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Triple-negative breast cancer (TNBC) is a heterogeneous disease and lacks effective treatment. Our previous study classified TNBCs into four subtypes with putative therapeutic targets. Here, we report the final results of FUTURE, a phase II umbrella trial designed to explore whether the subtyping-based strategy may improve the outcomes in metastatic TNBC patients. A total of 141 patients with a median of three previous lines of therapies in the metastatic setting were enrolled in seven parallel arms. Confirmed objective responses were achieved in 42 patients (29.8%; 95% confidence interval [CI], 22.4–38.1). The median values of progression-free survival and overall survival were 3.4 (95% CI: 2.7–4.2) and 10.7 (95% CI: 9.1–12.3) months, respectively. Given Bayesian predictive probability, efficacy boundaries were achieved in four arms. Furthermore, integrated genomic and clinicopathological profiling illustrated associations of clinical and genomic parameters with treatment efficacy, and the efficacy of novel antibody–drug conjugates was explored in preclinical TNBC models of subtypes for which treatment was futile. In general, the FUTURE strategy recruits patients efficiently and provides promising efficacy with manageable toxicities, outlining a direction for further clinical exploration.

With rare genetic variations having been increasingly recognized at a preclinical stage, a variety of early-phase clinical trials have been launched. Due to the low incidence rate of these variations, although the sample size of trials are small, it still needs a large number of patients for screening. With the advent of next-generation sequencing (NGS), multiple genetic variations can be detected simultaneously. Multiple biomarkers and agents can be evaluated using umbrella clinical trials, which rapidly and effectively screen and enroll patients for parallel sub-studies using NGS.We designed an open-label, multi-center, phase II clinical trial CTONG1702. This is an adaptive umbrella trial that will evaluate the efficacy and safety of several biomarker-driven agents, including tyrosine kinase inhibitors (TKIs) and a PD-1 inhibitor, in stage IIIB to IV patients (eighth AJCC) with non-small-cell lung cancer (NSCLC) patients. Patients will be enrolled in parallel sub-studies based on the results of NGS and PD-L1 IHC analysis. Patients who are not eligible for CTONG1702 will be enrolled in the observational real-world study CTONG1705. This study aims to develop a large-scale genomic database and explore the relationship between genetic variations in NSCLC patients and clinical outcomes.The adaptive umbrella trial will evaluate multi-targets and multi-drugs in advanced NSCLC patients (CTONG1702). In addition, the simultaneously initiated real-world study will provide additional data for clinical practice (CTONG1705).

The outbreak of coronavirus disease 2019 (COVID-19) is a significant challenge for clinicians, especially for immunocompromised cancer patients. By analyzing the impact of COVID-19 on the immune microenvironment of colorectal cancer (CRC) patients at the tissue level and single-cell level, we found that CRC patients are more easily infected by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), but promotion of infiltration and differentiation of monocytes makes them more likely to develop severe COVID-19. Because of the continuing activation of nuclear factor (NF)-κB and C-C chemokine receptor type 5 (CCR5) signaling pathways in monocytes, imbalance of macrophage polarization can aggravate the cytokine release syndrome. Therefore, regulating the infiltration and differentiation of monocytes is helpful for the treatment of COVID-19 in CRC patients.

Background An increasing number of studies have shown that iron, one of the indispensable trace elements in the human body, is closely related to the occurrence and development of cancer. However, few studies have clearly demonstrated the role of the iron levels in lung cancer patients, or the potential effects of inflammation on iron levels. Methods The clinical data for lung cancer patients and non-lung cancer participants were retrospectively analyzed. The serum iron and ferritin levels were measured and compared using a rank-sum test. The correlation between the serum iron/ferritin and C-reactive protein (CRP) was analyzed by rank correlation. The cut-off values for continuous variables were obtained by the receiver operating characteristic curve (ROC) method. An analysis of potential prognostic factors in lung cancer patients was conducted by univariate and multivariate survival analyses. Results The serum iron levels in patients with extensive small-cell lung cancer (SCLC) were lower than those with limited-stage SCLC, and the levels of serum ferritin and CRP in those with extensive SCLC were higher than those with limited-stage SCLC. Similarly, the serum iron levels in patients with stage IV non-small cell lung cancer (NSCLC) were lower than those of patients with stage Ⅰ-Ⅲ disease, and the levels of serum ferritin and CRP in those with stage IV NSCLC were higher than those in stages Ⅰ-Ⅲ. The serum iron level was negatively correlated with the level of CRP, while the serum ferritin level was positively correlated with CRP. The stage of lung cancer, but not the serum iron/ ferritin level, was an independent prognostic factor in lung cancer patients. Conclusions The serum iron and ferritin levels are associated with the staging of lung cancer. The later stages of lung cancer are associated with a lower serum iron level, a higher serum ferritin level, and a higher CRP level. Inflammation may play an important role in regulating the serum iron and ferritin levels in lung cancer patients.

Objective There is strong evidence that the body composition can affect the progression-free survival (PFS) and overall survival (OS) in patients with a variety of cancers. The main objective of this study was to investigate the effect of body composition on the prognosis of patients with advanced gastrointestinal and colorectal cancers who received first-line palliative chemotherapy. Methods Patients who were newly-diagnosed with advanced gastrointestinal or colorectal cancer and received standard first-line palliative chemotherapy from January 2017 to December 2018 were included in this retrospective study. An analysis of computed tomography images was performed to determine the skeletal muscle index (SMI), which reflects the skeletal muscle mass and skeletal muscle density (SMD) related to muscle strength. A Kaplan-Meier survival analysis and log-rank test were used to compare the survival relationships among groups stratified by the SMI, and a Cox proportional hazard model was used for a multivariate analysis. Results A total of 108 patients met the inclusion criteria, including 41 cases of gastric cancer, 46 cases of left colorectal cancer, and 21 cases of right colon cancer. In patients with gastric cancer, the OS of women was significantly shorter than that of men. The OS of patients with a low SMI, low SMD, and low phase angle (PA) was significantly shorter than that of patients with high values (P ≤ 0.05). In the multivariate analysis, the SMD was significantly associated with the patients' long-term survival [Hazard Ratio (HR) = 0.904, 95% CI: 0.840~0.974, P = 0.008]. For patients with a low SMI and PA, the PFS was significantly shorter than that of patients with high values (P ≤ 0.05). In patients with left colon cancer, the PA and SMD were both independent risk factors for a poorer long-term prognosis (HR = 0.375, 95% CI: = 0.167~0.840, P = 0.017; HR = 0.887, 95% CI: 0.824~0.954, P = 0.001). Among right colon cancer patients, the PFS and OS of those with a low SMD were significantly lower than those for patients with high values (P ≤ 0.05). Conclusion The PA is an independent risk factor for the OS of left colon cancer patients; the SMD is an independent risk factor for the survival of patients with gastric cancer, left colon cancer, and right colon cancer.

Circulating tumor DNA (ctDNA) in the bloodstream can be used to reliably identify a minimal residual disease (MRD). ctDNA-MRD has demonstrated clinical values as a predictive and prognostic marker for resectable non-small cell lung cancer (NSCLC) regarding efficacy evaluation, recurrence monitoring, risk classification, and adjuvant treatment choices, and it has the advantage of being non-invasive, real-time, and dynamic. A recent large-scale prospective study of patients with resectable NSCLC revealed that patients with longitudinal undetectable MRD might represent a potentially curable population, benefiting many patients by eliminating wasteful therapies and side effects. However, there are still barriers to using ctDNA-MRD in clinical management, and the most significant is the lack of high-sensitivity detection technologies and consistent detection times. Herein, we defined the clinical significance of ctDNA-MRD in resectable NSCLC, summarized the available next-generation sequencing (NGS) detection approaches, and speculated on future clinical trial design and detection technology optimization.

Objective: There is no standard care for advanced non–small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutation in the third line. Our study aimed to assess the efficacy and safety of gefitinib as a third-line re-challenge treatment for advanced NSCLC patients with EGFR mutation. Materials and Methods: It was a multicenter, open-label, single-arm, phase II study. Stage IIIB/IV NSCLC patients with EGFR exon 19del/L858R mutation, who had benefited from first-line gefitinib treatment followed by second-line chemotherapy, received gefitinib 250 mg/d. The primary objective was disease control rate (DCR) at week 8. Results: Predefined DCR was achieved in 69.8% (95% confidence interval, 49.87-74.91) patients and objective response rate was reported in 4.7% (95% confidence interval, 0.78-13.06) patients. Median progression-free survival (PFS) was 4.4 months and overall survival (OS) was 10.3 months. Baseline T790M-negative patients achieved favorable DCR compared with T790M-positive patients (78.1% vs. 45.5%, P=0.0418), significantly longer median PFS (4.7 vs. 2.0 mo, P=0.0009) and median OS (15.2 vs. 7.7 mo, P=0.0132). We observed a negative correlation of PFS (r=−0.4396, P=0.0032), and OS (r=−0.3630, P=0.0167) with mutation abundance of exon 19del/L858R at baseline. Conclusions: Re-challenge with gefitinib is effective and could be a choice for third-line patients after the first-line EGFR-TKI treatment and second-line chemotherapy, especially for the T790M-negative patients.

Introduction BCR-ABL1 mutations require consideration during second-line tyrosine kinase inhibitor selection for patients with chronic myeloid leukemia (CML). The present retrospective analysis compared the frequency of BCR-ABL1 mutations in Asian and white patients in whom imatinib therapy had failed. Patients and Methods A nonstudy cohort (76 Asian patients from community clinical practices) and 2 study cohorts (29 Asian and 352 white patients from dasatinib phase II and III clinical trials) were identified. Results In the nonstudy cohort, 80 mutations were identified; the most frequent was T315I (15%), followed by phosphate-binding loop mutations E255K (11%), G250E (10%), and Y253H (10%). Asian patients had a greater proportion of T315I and phosphate-binding loop mutations compared with the white patients. The nonstudy cohort was less likely to have multiple mutations compared with either study cohort. Single mutations highly resistant to dasatinib, nilotinib, and bosutinib were more frequent in the Asian than in the white cohorts. Conclusion These results suggest that mutational analysis findings will be invaluable for choosing an appropriate second-line tyrosine kinase inhibitor in Asia.

Earlier studies on elucidating the role of lymphocytes in tumor immunity predominantly focused on T cells. However, the role of B cells in tumor immunity has increasingly received better attention in recent studies. The B cells that infiltrate tumor tissues are called tumor-infiltrating B cells (TIBs). It is found that TIBs play a multifaceted dual role in regulating tumor immunity rather than just tumor inhibition or promotion. In this article, latest research advances focusing on the relationship between TIBs and tumor complexity are reviewed, and light is shed on some novel ideas for exploiting TIBs as a possible tumor biomarker and potential therapeutic target against tumors.

MDA7/IL24 is a member of the IL-10 gene family that functions as a cytokine. Notably, supra-physiological endogenous MDA7 levels have been indicated to suppress tumor growth and induce apoptosis in different cancer types. In the present study, MDA7 roles were investigated during the proliferation of hepatocellular carcinoma (HCC) cells and the molecular mechanisms underlying this process. A lentiviral vector expressing MDA7/IL24 (LV-MDA7/IL24) was constructed and used to infect HCC SMMC-7721 cells. The expression levels of MDA7/IL24 in these cells were determined using RT-qPCR and western blot analysis. The effects of LV-MDA7/IL24 on cell proliferation were analyzed using MTT and colony formation assays. Furthermore, the influence of LV-MDA7/IL24 on cell apoptosis and cell cycle distribution were detected using flow cytometry. The underlying molecular mechanisms were investigated using microarray and western blot analysis. The expression of MDA7/IL24 was confirmed to be significantly increased in the cells infected with LV-MDA7/IL24 compared with that the negative-control infected group. Lentivirus-mediated MDA7/IL24 expression was found to inhibit HCC cell proliferation and colony formation, and it also induced cell arrest and apoptosis. Microarray analysis and western blotting results indicated that multiple cancer-associated pathways and oncogenes are regulated by MDA7/IL24, including cell cycle regulatory and apoptosis activation pathway. In conclusion, it was determined that MDA7/IL24 inhibits the proliferation and reduces the tumorigenicity of HCC cells by regulating cell cycle progression and inducing apoptosis, indicating that it may be used as a potential prognostic and therapeutic target in HCC.

The diagnosis and treatment of lung cancer have evolved into the era of precision medicine. Liquid biopsy, a minimally invasive approach, has emerged as a promising practice in genetic profiling and monitoring of lung cancer. Translating liquid biopsy from bench to bedside has encountered various challenges, including technique selection, protocol standardisation, data analysis and cost management. Regarding these challenges, the 2016 Chinese Lung Cancer Summit expert panel organised a trilateral forum involving oncologists, clinicians, clinical researchers, and industrial expertise on the 13th Chinese Lung Cancer Summit to formally discuss these controversies. Six consensuses were reached to guide the use of liquid biopsy and perform precision medicine in both clinic and research.

Objectives To explore NPM1, FLT3-ITD, CEBPA, and c-kit mutations in patients with acute myeloid leukemia (AML) from Chinese population. Methods In this study, we retrospectively analyzed the prevalence and clinical profile of NPM1, FLT3-ITD, CEBPA, and c-kit mutations in 312 patients with de novo AML. Results The frequencies of NPM1, FLT3-ITD, c-kit, and CEBPA mutations were 15.4, 14.0, 7.64, and 25.6%, respectively. The occurrence rate of NPM1 mutations increased with age in patients younger than 60 years. NPM1, c-kit, and CEBPA mutations were all associated with French-American-British subtypes. Patients with NPM1 mutations and FLT3-ITD presented with higher peripheral white blood cell counts and marrow blast percentages. Conclusion Both this and previous studies may suggest low frequencies of NPM1 and FLT3-ITD mutations in AML patients from the Chinese population, and they may have a synergistic function in stimulating proliferation of leukemia cells.

Certain molecular mutations are associated with signs of cell morphology and differentiation in acute myeloid leukemia (AML). However, only limited data are available for the detailed analysis of such correlations. In this study, AML patients were classified into 4 subsets according to CD34, HLA-DR and CD11c expression levels. Significantly low CD34 antigen expression was observed in nucleophosmin (NPM1)-mutated patients and in those with FMS-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD). No correlations were observed among NPM1 mutations, FLT3-ITD and monocytic morphology in patients without CD34 expression. Both NPM1 mutations and FLT3-ITD were absent in cluster IIb patients (CD34+CD11c-). The associations among NPM1 mutations, FLT3-ITD and the surface molecular signature of leukemic cells may offer beneficial information about the pathogenesis of AML.

e15044Background: Preclinical studies showed that antibody-dependent cell-mediated cytotoxicity mediated by CD56+natural killer (NK) cells contributed to the activity of rituximab (R) in patients (...

e20504Background: There are significantly differences in the efficacy of platinum based chemotherapy in NSCLC patients. Platinum leads to the cell death by damaging DNA through the chain cross link...

CCAAT/enhancer binding protein alpha gene (CEBPA) is an important transcription factor in maintenance of differentiation of granulocyte series of hematopoietic system. It plays a key role in regulating cell proliferation and differentiation. CEBPA mutation easily occurs in M1 and M2 type of acute myeloid leukemia, about 5%-14% in adult acute myeloid leukemia and 7.9% in children with acute myeloid leukemia. At present, domestic CEBPA mutation research is far less than abroad. This review focuses on the structual characteristics and detection method of CEBPA, CEBPA clinical features, the effect of CEBPA mutation on the prognosis of patients and the choice of treatment.

Low-dose computed tomography (LDCT) has been increasingly accepted as an efficient screening method for high-risk individuals to reduce lung cancer mortality. However, there remains a gap of knowledge in the practical implementation of screening on a larger scale, especially for low-risk individuals. The aim of this study is to initiate discussion through an evidence-based analysis and provide valuable suggestions on LDCT screening for lung cancer in clinical practice. Among previously published randomized controlled trials (RCTs), the National Lung Screening Trial (NLST) is the only one demonstrating positive results in a high-risk population of old age and heavy smokers. It is also shown that the potential harms include false-positive findings, radiation exposure etc., but its magnitude is uncertain. In the meantime, the current risk stratification system is inadequate, and is difficult to define selection criteria. Thus, the efficacy of LDCT in lung cancer screening needs to be confirmed in future trials, and the procedure should not be proposed to individuals without comparable risk to those in the NLST. Furthermore, there is a lack of evidence to support the expansion of LDCT screening to low-risk individuals. Therefore, recommendation of LDCT screening for these patients could be premature in clinical practice although some of them might be missed based on current definition of risk factors. Further studies and advances in risk assessment tools are urgently needed to address the concerns about lung cancer screening in order to improve the outcomes of lung cancer.

To screen serum biomarkers for minimal residual disease (MRD) monitoring according to differential peptidomics profile in the serum from the patients with acute leukemia (AL) and healthy controls.Serum polypeptides from 90 AL patients, 60 healthy controls and 20 patients with benign hematological disorders were enriched by copper chelate magnetic beads, and the peptidomics profile was obtained by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis. And the intensities of differential peptides were calculated to assess MRD level.The diagnostic models by using support vector machine (SVM) algorithm according to differential peptides between AL patients and healthy controls with P0.01 by t-test were established. The sensitivity and specificity of distinguishing AL patients from healthy controls were 98% and 99%, respectively. The model obtained a sensitivity of 98% and a specificity of 96% for distinguishing newly-diagnosed AL patients from AL patients with hematological complete remission (AL-HCR). Then a sensitivity of 92% and a specificity of 93% were obtained for distinguishing patients with AL-CR from AL patients with molecular complete remission (AL- MR). The intensity of peptide with m/z (mass-to-charge ratio) 4468 was significantly higher in newly- diagnosed AL patients compared to healthy controls, and gradually decreased with the increase of remission degree, and it was not found increase in patients with benign hematological disorders.The SVM diagnostic model established by differential serum peptide profile could be used to discriminate AL patients with different stages of remission and to evaluate the treatment efficacy. The peptide of m/z 4468 could be used for MRD assessment, and continuous monitoring of its expression level will play an important role in the individual treatment and recurrence prediction.

This study was aimed to investigate the correlation of NPM1 and FLT3-ITD mutations with leukocyte count in peripheral blood and bone marrow blasts in patients with acute myeloid leukemia (AML). Fifty-one acute myeloid leukemia patients with normal karyotype from January 2009 to December 2011 were enrolled in this study. The clinical data of 51 cases were analyzed retrospectively. Out of 52 cases 22 were male, and 29 were female. The median age was 47 years old (ranged from 14 to 83 years old). The de novo patients were examined by bone marrow cytomorphology and blood routine analysis. Polymerase chain reaction was used to analyze the NPM1 and FLT3-ITD mutations. The results showed that the patients with NPM1 mutations had higher leukocyte count compared with those without mutations (30.7×10(9)/L vs 8.6×10(9)/L, P = 0.002). FLT3-ITD mutation was related to higher leukocyte count (42.38×10(9)/L vs 11.45×10(9)/L without mutation, P = 0.033) and blasts (74.0% vs 60.25% without mutation, P = 0.036). The leukocyte count and percentage of bone marrow blasts were lowest in the patients with neither mutations, and gradually increasing in the NPM1(-) mutation, FLT3-ITD(-) mutation, and NPM1(+) mutation, FLT3-ITDI(+) mutation, and NPM1(+)/FLT3-ITD(+) mutation groups (P0.05). The patients tended to have NPM1 (P = 0.002) and FLT3-ITD (P = 0.033) mutations when their leukocyte counts were more than 12.55×10(9)/L and 37.85×10(9)/L, respectively. Those with bone marrow blast more than 72.25% showed higher rate of FLT3-ITD mutation (P = 0.008). Patients with NPM1 mutations had higher complete remission rate than those without NPM1 mutation (78.13% vs 40.0%, χ(2) = 4.651, P = 0.031) after remission induction therapy. It is concluded that both NPM1 and FLT3-ITD mutations are linked to higher leukocyte count and blast percentage, suggesting that both mutations may be associated with increased proliferation of leukemia cells, and may have a synergistic function in stimulating proliferation.

To analyze the cytogenetic characteristics of different age subgroups in patients with acute myeloid leukemia (AML), and to explore the relationship between age and cytogenetics.Between January 2004 and December 2011, Bone marrow (BM) samples from 640 patients with de novo AML were analyzed retrospectively. The analyses were performed according to standard culturing and banding techniques, and clonal abnormalities were defined and described according to the International System for Human Cytogenetic Nomenclature (ISCN 2009). The cytogenetic subtypes were performed as normal, balanced, and unbalanced karyotypes. In the last group, the age distribution of complex and monosome karyotypes were further analyzed. The patients were divided into 8 age groups: 0 - 9, 10 - 19, 20 - 29, 30 - 39, 40 - 49, 50 - 59, 60 - 69, and ≥ 70 year old groups.The distribution of normal, balanced, and unbalanced karyotypes showed age specific characteristics. The incidence of normal karyotype increased from 6.67% (0 ∼ 9 year old) to 58.33% (≥ 70) (χ(2) = 20.68, P = 0.001) and balanced karyotype decreased from 73.33% (0 ∼ 9) to 11.11% (≥ 70) (χ(2) = 48.22, P0.01). The frequency of unbalanced karyotypes increased from 20.0% (0 ∼ 9) to 30.56% (≥ 70) (χ(2) = 18.963, P = 0.008). The frequency of complex karyotype was 6.67% in 0 - 9 year old group, followed by 0% in 10 - 19 and 20 - 29 year old group, and from 1.72% to 11.11% from 30 - 39 to ≥ 70 year old group (χ(2) = 8.341, P = 0.08). Monosome karyotype was only detected in patients in 30 year old or older groups. Although an increased tendency was observed with ages, there was no significant difference (χ(2) = 4.778, P = 0.311).The different age profiles of the cytogenetic subtypes may indicate the different mechanisms of the pathogenesis of AML, which may also offer beneficial information for etiological research of AML.

Certain molecular mutations are associated with signs of cell morphology and differentiation in acute myeloid leukemia (AML). However, only limited data are available for the detailed analysis of such correlations. In this study, AML patients were classified into 4 subsets according to CD34, HLA-DR and CD11c expression levels. Significantly low CD34 antigen expression was observed in nucleophosmin (NPM1)-mutated patients and in those with FMS-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD). No correlations were observed among NPM1 mutations, FLT3-ITD and monocytic morphology in patients without CD34 expression. Both NPM1 mutations and FLT3-ITD were absent in cluster IIb patients (CD34(+)CD11c(-)). The associations among NPM1 mutations, FLT3-ITD and the surface molecular signature of leukemic cells may offer beneficial information about the pathogenesis of AML.

To find a kind of quick and effective haemostasis to decrease the mortality of severe bleeding.18 severe bleeding patients with different cause received recombinant activated factor VIIa (rFVIIa) were analyzed retrospectively.Of total 18 cases with severe bleeding, 13 cases cured, 3 cases were effective, 2 cases ineffective. The total clinical effective rate is 88.89%. After using rFVIIa, the PT, APTT and fibrinogen level of 6 DIC patients returned to normal within 12 hours; 13 patients whose the amount of bleeding can be evaluated stopped bleeding quickly. The fastest onset time was 10 min.rFVIIa can stanch severe bleeding for a variety of reasons rapidly and effectively, including coagulopathy, thrombocytopenia, and obstetric hemorrhage. Application of rFVIIa may decrease mortality, when conventional treatment is not valid.

The activation of Fli-1, an Ets transcription factor, is the critical genetic event in Friend murine leukemia virus (F-MuLV)–induced erythroleukemia. Fli-1 overexpression leads to erythropoietin-dependent erythroblast proliferation, enhanced survival, and inhibition of terminal differentiation, through activation of the Ras pathway. However, the mechanism by which Fli-1 activates this signal transduction pathway has yet to be identified. Down-regulation of the Src homology 2 (SH2) domain-containing inositol-5-phosphatase-1 (SHIP-1) is associated with erythropoietin-stimulated erythroleukemic cells and correlates with increased proliferation of transformed cells. In this study, we have shown that F-MuLV–infected SHIP-1 knockout mice display accelerated erythroleukemia progression. In addition, RNA interference (RNAi)-mediated suppression of SHIP-1 in erythroleukemia cells activates the phosphatidylinositol 3-kinase (PI 3-K) and extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathways, blocks erythroid differentiation, accelerates erythropoietin-induced proliferation, and leads to PI 3-K–dependent Fli-1 up-regulation. Chromatin immunoprecipitation and luciferase assays confirmed that Fli-1 binds directly to an Ets DNA binding site within the SHIP-1 promoter and suppresses SHIP-1 transcription. These data provide evidence to suggest that SHIP-1 is a direct Fli-1 target, SHIP-1 and Fli-1 regulate each other in a negative feedback loop, and the suppression of SHIP-1 by Fli-1 plays an important role in the transformation of erythroid progenitors by F-MuLV.

This study was aimed to construct the shRNA eukaryotic expression vectors of M2-pyruvate kinase gene (pkm2) and to investigate the effects of pkm2 gene interference on the drug resistance of acute promyelocytic leukemia (APL) cells in vitro. Three specific shRNAs of pkm2 gene were designed and cloned into PBSU6 vector containing a U6 promotor. The constructed plasmids were identified and proved by the restriction sequence analysis. Then the effect of pkm2-shRNA on the protein expression of endogenous PKM2 was detected in NB4R2 cells, a drug resistant cell line of APL by Western blot. The alteration of NB4R2 cell differentiation with the interference of pkm2 gene was also validated by nitroblue tetrazolium (NBT) reduction test. The results showed that three specific shRNA eukaryotic expression vectors targeting pkm2 were successfully constructed. The efficiency of pkm2 gene silence was proved at protein level. The interference of pkm2 gene could significantly enhance the cell differentiation in the drug resistant NB4R2 cell line. It is concluded that the DNA vector containing pkm2 targeting shRNA remarkably promotes the differentiation of NB4R2 cells, showing the prospects of developing the gene target drug.

MicroRNAs (miRNAs) are a newly discovered class of posttranscriptional regulatory noncoding small RNAs. Recent evidence has shown that miRNA misexpression correlates with progression of various human cancers. Friend erythroleukemia has been used as an excellent system for the identification and characterization of oncogenes and tumor suppressor genes involved in neoplastic transformation. Using this model, we have isolated a novel integration site designated Fli-3, from a Friend murine leukemia virus (F-MuLV)–induced erythroleukemia. The Fli-3 transcription unit is a murine homologue of the human gene C13orf25 that includes a region encoding the mir-17–92 miRNA cluster. C13orf25 is the target gene of 13q31 chromosomal amplification in human B-cell lymphomas and other malignancies. The erythroleukemias that have acquired either insertional activation or amplification of Fli-3 express higher levels of the primary or mature miRNAs derived from mir-17–92. The ectopic expression of Fli-3 in an erythroblastic cell line switches erythropoietin (Epo)–induced differentiation to Epo-induced proliferation through activation of the Ras and PI3K pathways. Such a response is associated with alteration in the expression of several regulatory factors, such as Spi-1 and p27 (Kip1). These findings highlight the potential of the Fli-3 encoding mir-17–92 in the development of erythroleukemia and its important role in hematopoiesis.

To explore the molecular mechanisms of apoptotic NB4 cells induced by the histone deacetylase inhibitor, sodium butyrate(SB).SB was exposed to NB4 cells at a final concentration of 1.0 mmol/L. The untreated and treated cells were analysed with FACS and 2-dimensional electrophoresis (2-DE) at 0, 12, 24, 48 and 72 h. The changed protein spots were identified with MALDI-TOF-MS and ESI-TOF-MS/MS.SB induced apoptosis of NB4 cells. Twenty-one changed proteins involving apoptotic signal transduction, immunological regulation, transcriptional control, cellular metabolism, molecular transport and so on were identified. Thirteen of them had been reported to be related to apoptosis.SB can induce apoptosis and many functional protein changes in tumor cells. These results pave the way to further explore the anti-tumor mechanism of SB.

This study was aimed to analyze the different proteomes between human acute leukemia (AL) cells and normal white blood cells by proteomic technology in order to lay the basis for diagnosing AL and understanding the mechanism of leukemogenesis. The proteins from AL cells of 40 AL patients identified by FAB classification and proteins from normal lymphocytes and granulocytes of 20 normal volunteers were separated by two-dimensional electrophoresis (2-DE), and the differentially expressed proteins between the two groups were identified by both matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) and electronspray ionization (ESI)-MS/MS. The results showed that among the differentially expressed proteins between AL cells and normal lymphocytes and granulocytes, some proteins involved in the process of malignant transformation (such as Op18, NM23-H1), cell proliferation (such as PCNA) and apoptosis inhibition (such as tumor necrosis factor inhibitor protein) were found to be up regulated in AL cells. However, some proteins involved in differentiation and physiological functions of normal cells were down regulated in AL cells. It is concluded that there are many events involved in the process of leukemogenesis, expression of some proteins relating to the malignant transformation, cell proliferation and apoptosis inhibition are up-regulated in AL cells. The proteome analysis may provide a new approach to explaining the molecular mechanism underlying the pathogenesis of AL.

To observe the therapeutic efficacy and side effects of arsenic trioxide (As2O3)combined with low-dose all-trans retinoic acid (ATRA) on remission induction in newly-diagnosed and relapsed patients with acute promyeloeytic leukemia (APL).224 patients of APL, 156 newly diagnosed patients, aged 34 (13 approximately 62), with a male/female ratio of 1.56, and 28 relapsed patients, aged, aged 34 (12 approximately 63), with a male/female ratio of 1.89, underwent As2O3 + ATRA therapy. The therapeutic effects was compared with that of As2O3 alone treatment on 40 newly diagnosed patients and 25 relapsed patients and that of ATRA alone treatment on 36 newly diagnosed patients and 15 relapsed patients. The treatment protocol for the combination group was as following: As2O3 was administered intravenously at a dose of 10 mg/day and ATRA was given orally three times per day at a dose of 10 mg. The complete remission (CR) rate, period to CR, incidence of early death and side effects were observed in the three groups.In the newly-diagnosed patients, there was no significant difference in CR rate among the three groups (92.5% for the As2O3/LD-ATRA group, 83.8% for the ATRA group, and 90% for the As2O3 group respectively). In comparison with As2O3 alone, administration of LD-ATRA to the patients in the As2O3/LD-ATRA group significantly shortened the period to CR (the medium time to CR was 28 days for the As2O3/LD-ATRA group and 39 days for the As2O3 group respectively). As compared to ATRA alone, treatment with As2O3 with low-dose ATRA showed a significantly lower incidence of early death (2.5% for the As2O3/LD-ATRA group and 13.9% for the ATRA group respectively). In the relapsed patients, the CR rate was significantly higher in the group treated with As2O3/LD-ATRA (71.4% for the As2O3/LD-ATRA group, 32.0% for the ATRA group, and 43.0% for the As2O3 group respectively). The combined use of LD-ATRA with As2O3 did not further enhance toxic side effects as compared to As2O3 alone or ATRA alone.As2O3/LD-ATRA regimen is superior to either regimen given alone to patients with APL. It is an efficient therapeutic approach to APL patients using a combination of As2O3 with low-dose ATRA.

French-American-British (FAB) classification of acute leukemia with genetic heterogeneity is important for treatment and prognosis. However, the distinct protein profiles that contribute to the subtypes and facilitate molecular definition of acute leukemia classification are still unclear.The proteins of leukemic cells from 61 cases of acute leukemia characterized by FAB classification were separated by two-dimensional electrophoresis, and the differentially expressed protein spots were identified by both matrix-assisted laser desorption/ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) and tandem electrospray ionization MS (ESI-MS/MS).The distinct protein profiles of acute leukemia FAB types or subtypes were successfully explored, including acute myeloid leukemia (AML), its subtypes (M2, M3, and M5) and acute lymphoid leukemia (ALL), which were homogeneous within substantial samples of the respective subgroups but clearly differed from all other subgroups. We found a group of proteins that were highly expressed in M2 and M3, rather than other subtypes. Among them, myeloid-related proteins 8 and 14 were first reported to mark AML differentiation and to differentiate AML from ALL. Heat shock 27 kDa protein 1 and other proteins that are highly expressed in ALL may play important roles in clinically distinguishing AML from ALL. Another set of proteins up-regulated was restricted to granulocytic lineage leukemia. High-level expression of NM23-H1 was found in all but the M3a subtype, with favorable prognosis.These data have implications in delineating the pathways of aberrant gene expression underlying the pathogenesis of acute leukemia and could facilitate molecular definition of FAB classification. The extension of the present analysis to currently less well-defined acute leukemias will identify additional subgroups.

Acute myelogenous leukemia (AML) cells are organized in a hierarchical fashion, with only the most primitive rare population (leukemia stem cell, LSC) of AML cells capable of maintaining the leukemic clone. A broad range of studies has indicated that AML results from mutations at the level of the stem cells of AML cells. The changes of cellular and molecular features in these malignant stem cells determine the features of leukemic clone and give rise to different subtypes of AML. LSCs share some similar characteristics with normal hematopoietic stem cells (HSC) including the ability to self-renew, and also have the potential of limited differentiation. LSCs, also have some features that are not found in normal HSC. LSCs have unique phenotype such as CD90-, CD117- and CD123+. Tumor-suppressor protein-death associated protein kinase and interferon regulatory factor 1 were overexpressed in LSCs, but not in normal HSC. Due to a predominantly G0 cell-cycle status, LSCs may not be responsive to conventional chemotherapeutic agents, compared with leukemia blasts. It is proposed that surviving LSCs are a major contributing factor to leukemic relapse. Although LSC population is likely to be drug-resistant, quiescent LSCs are preferentially susceptible to apoptosis induction while sparing normal HSC, with the appropriate stimulus such as proteasome inhibitor MG-132. This article reviewed the data emerging from the study of LSCs, and elucidated the distinct cellular and molecular characteristics of the LSC population, which may shed new light on AML therapy and leukemogenesis study.