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Anterior gradient 2 protein belongs to a family of chaperone‐like proteins, namely protein disulfide isomerase. Generally, AGR2 is highly expressed in mucus‐secreting cells and endocrine organs, and in this study, we aimed to evaluate AGR2 and cell cycle molecules in epithelial ovarian cancer and its implications on prognosis. One hundred seventy‐five patient's samples that were diagnosed with primary epithelial ovarian carcinoma were selected. All the patients were treated with platinum‐taxane standard chemotherapy after surgery and CA125 serum levels were routinely determined. Four‐micrometer‐thick sections were processed by immunohistochemistry using an automated immunostainer, Ventana BenchMark AutoStainer with AGR2, cyclin D1, p21WAF1, and p53. Forty‐nine of 167 cases (29.3%) showed strong to moderate cytoplasmic marking of AGR2, and 118 (70.7%) had weak to negative expression. The absence of the AGR2 protein was observed in high‐grade serous carcinoma (P

Graphical abstract Image 1, Prevalence of Kaposi sarcoma-associated herpesvirus (KSHV/HHV-8) varies greatly in different populations. We hypothesized that the actual prevalence of KSHV/HHV8 infection in humans is underestimated by the currently available serological tests. We analyzed four independent patient cohorts with post-surgical or post-chemotherapy sepsis, chronic lymphocytic leukemia and post-surgical patients with abdominal surgical interventions. Levels of specific KSHV-encoded miRNAs were measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and KSHV/HHV-8 IgG were measured by immunoassay. We also measured specific miRNAs from Epstein Barr Virus (EBV), a virus closely related to KSHV/HHV-8, and determined the EBV serological status by ELISA for Epstein-Barr nuclear antigen 1 (EBNA-1) IgG. Finally, we identified the viral miRNAs by in situ hybridization (ISH) in bone marrow cells. In training/validation settings using independent multi-institutional cohorts of 300 plasma samples, we identified in 78.50% of the samples detectable expression of at least one of the three tested KSHV-miRNAs by RT-qPCR, while only 27.57% of samples were found to be seropositive for KSHV/HHV-8 IgG (P, Highlights • There is no agreement on a standard assay to detect the true prevalence of Kaposi sarcoma-associated herpesvirus (KSHV) infection. • Measurement of the viral miRNAs in plasma by RT-qPCR allows a direct and accurate assessment of viral infection. • Measurement of the viral miRNAs in plasma by RT-qPCR shows prevalence of KSHV infection in immuno-depressed patients. • Measurement of plasma viral miRNAs for viral infection assessment has the potential to become a “gold” standard method in the clinical practice. Chronic viral infections represent risk factors for diseases and development of infection-related complications. There is no agreement on a standard assay to detect the true prevalence of Kaposi sarcoma-associated herpesvirus (KSHV) infection. The current method used in the clinical practice (ELISA-test) identifies a great geographic variation in KSHV seroprevalence and may underestimate the true-prevalence of KSHV infection. Here we showed that detection of plasma viral miRNAs levels for the identification of viral infection (e.g., KSHV, Epstein-Bar virus or EBV) is more accurate than the current method for detection of virus-derived antigen, especially in patients with low number of immune cells.

Myelofibrosis (MF) is a myeloproliferative neoplasm characterized by cytopenia and extramedullary hematopoiesis, resulting in splenomegaly. Multiple pathological mechanisms (e.g., circulating cytokines and genetic alterations, such as JAKV617F mutation) have been implicated in the etiology of MF, but the molecular mechanism causing resistance to JAK2V617F inhibitor therapy remains unknown. Among MF patients who were treated with the JAK inhibitor ruxolitinib, we compared noncoding RNA profiles of ruxolitinib therapy responders versus nonresponders and found miR-543 was significantly upregulated in nonresponders. We validated these findings by reverse transcription–quantitative PCR. in this same cohort, in 2 additional independent MF patient cohorts from the United States and Romania, and in a JAK2V617F mouse model of MF. Both in vitro and in vivo models were used to determine the underlying molecular mechanism of miR-543 in MF. Here, we demonstrate that miR-543 targets the dioxygenases ten-eleven translocation 1 (TET1) and 2 (TET2) in patients and in vitro, causing increased levels of global 5-methylcytosine, while decreasing the acetylation of histone 3, STAT3, and tumor protein p53. Mechanistically, we found that activation of STAT3 by JAKs epigenetically controls miR-543 expression via binding the promoter region of miR-543. Furthermore, miR-543 upregulation promotes the expression of genes related to drug metabolism, including CYP3A4, which is involved in ruxolitinib metabolism. Our findings suggest miR-543 as a potentially novel biomarker for the prognosis of MF patients with a high risk of treatment resistance and as a potentially new target for the development of new treatment options The work in GAC’s laboratory was partly supported by NIH/National Center for Advancing Translational Sciences grant UH3TR00943-01, the NIH/National Cancer Institute (NCI) grant 1 R01 CA182905-01, U54 grant UPR/MDACC Partnership for Excellence in Cancer Research 2016 Pilot Project, Team Department of Defense grant CA160445P1, a Ladies Leukemia League grant, a Chronic Lymphocytic Leukemia Moonshot Flagship project, a Sister Institution Network Fund 2017 grant, and the Estate of C. G. Johnson, Jr. EFM was supported in part by award number P50 CA140388 from the NCI and by the NIH Clinical Research Loan Repayment Program. AMS was supported by Fundação para a Ciência e a Tecnologia through fellowship SFRH/BD/85968/2012. ZJZ’s work was supported by grants from the MPN Foundation and the Oklahoma Center for the Advancement of Science and Technology. DG, MPD, and IBN were supported in part by a Programul Opera¿ional Competitivitate grant nr35/01.09.2016, ID 37_796, titled “Clinical and economical impact of personalized targeted anti-mi-croRNA therapies in reconverting lung cancer chemoresistance” (CANTEMIR). NSDA was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo, BEPE 2016/09349-4.

// Beatriz de Melo Maia 1, 2 , Iara Santana Rodrigues 1 , Erica Mie Akagi 1 , Nayra Soares do Amaral 1 , Hui Ling 2 , Paloma Monroig 2 , Fernando Augusto Soares 1 , George Adrian Calin 2, 3 , Rafael Malagoli Rocha 4 1 Molecular Morphology Laboratory, Anatomic Pathology Department, AC Camargo Cancer Center, Sao Paulo, Brazil 2 Department of Experimental Therapeutics, The University of Texas, MD Anderson Cancer Center, Houston, TX, USA 3 The Center for RNA Interference and Non-Coding RNAs, The University of Texas, MD Anderson Cancer Center, Houston, TX, USA 4 Gynecology Laboratory, Gynecologic Department Federal University of Sao Paulo, Sao Paulo, Brazil Correspondence to: Rafael Malagoli Rocha, email: rafael.malagoli@gmail.com Keywords: vulvar cancer, microRNAs, cellular assays, hsa-miR-223-5p, TP63 Received: January 11, 2016 Accepted: May 08, 2016 Published: June 23, 2016 ABSTRACT MiR-223-5p has been previously mentioned to be associated with tumor metastasis in HPV negative vulvar carcinomas, such as in several other tumor types. In the present study, we hypothesized that this microRNA would be important in vulvar cancer carcinogenesis and progression. To investigate this, we artificially mimicked miR-223-5p expression in a cell line derived from lymph node metastasis of vulvar carcinoma (SW962) and performed in vitro assays. As results, lower cell proliferation ( p < 0.01) and migration ( p < 0.001) were observed when miR-223-5p was overexpressed. In contrast, increased invasive potential of these cells was verified ( p < 0.004). In silico search indicated that miR-223-5p targets TP63 , member of the TP53 family of proteins, largely described with importance in vulvar cancer. We experimentally demonstrated that this microRNA is capable to decrease levels of p63 at both mRNA and protein levels ( p < 0.001, and p < 0.0001; respectively). Also, a significant inverse correlation was observed between miR-223-5p and p63 expressions in tumors from patients ( p = 0.0365). Furthermore, low p63 protein expression was correlated with deeper tumor invasion ( p = 0.0491) and lower patient overall survival ( p = 0.0494). Our study points out miR-223-5p overexpression as a putative pathological mechanism of tumor invasion and a promising therapeutic target and highlights the importance of both miR-223-5p and p63 as prognostic factors in vulvar cancer. Also, it is plausible that the evaluation of p63 expression in vulvar cancer at the biopsy level may bring important contribution on prognostic establishment and in elaborating better surgical approaches for vulvar cancer patients.

To identify biomarkers that could predict response or lack of response to conventional chemotherapy at the time of diagnosis of high‑grade serous ovarian carcinoma (HGSOC), the present study compared large‑scale gene expression from patients with short or long disease‑free survival times, according to the last cycle of chemotherapy, and validated these findings using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and conventional immunohistochemical (IHC) analysis. Samples were selected for microarray evaluation, at the time of diagnosis, using the following criteria: Identical debulking primary surgery, International Federation of Gynaecology and Obstetrics staging, histological subtype and grade. These were divided into 2 groups, regarding the outcome after 2 years of follow-up. Prostaglandin D2 synthase 21 kDa (brain) (PTGDS) was found to be expressed at a significantly higher level in the tumours of patients with a short disease‑free survival time, and this was validated by RT‑qPCR in all samples. Furthermore, the study evaluated PGD2, the protein product of the PTGDS gene, in a large cohort of 114 HGSOC patients using the Ventana Benchmark automated platform, and IHC positivity was correlated with clinicopathological data and outcome. The global gene expression analysis identified 1,149 genes that were differentially expressed in microarray data, according to the patient outcome. Further analysis RT‑qPCR validated PTGDS gene expression in the same samples (r=0.945; P

Introduction: Bevacizumab is an antiangiogenic agent approved to treat ovarian cancer in combination to chemotherapy. The relative benefit of bevacizumab in ovarian cancer patients seems to be greater the more the disease becomes platinum resistant. Cyclin E1 overexpression is a marker of platinum resistance. In this study we aimed to evaluate the benefit of bevacizumab in platinum sensitive recurrent ovarian cancer and to test if the benefit changes according to platinum-free interval (PFI) and cyclin E1 expression. Methods: We retrospectively evaluated data from patients with platinum sensitive recurrent ovarian cancer treated with CT plus bevacizumab (Bev group) and CT alone (CT group) at a tertiary cancer center in Brazil from 2005 to 2017. The two groups were paired according to histology, platinum free interval and number of previous treatment lines. Cyclin E1 expression was evaluated by immunohistochemistry in tissue microarray. Progression-free survival (PFS) was compared between the groups with log rank test and cox regression. Results: 124 patients were included, 62 in each group. Bev group and CT group were well balanced regarding histology (high grade serous carcinoma 94% in Bev group, 93% in CT group), PFI (PFI > 12 months in 36.2% in Bev group, 39.3% in CT group) and number of previous treatment lines (one previous chemotherapy in 55.7% in Bev group, 60.0% in CT group). Median age and median PFI were 56.2 years old and 59.2 years old, and 9.7 months and 10.5 months, in the Bev and CT groups, respectively. All patients were treated with platinum doublets with paclitaxel, gemcitabine or liposomal doxorubicin except for one patient treated with cisplatin plus bevacizumab and one patient treated with liposomal doxorubicin. Median PFS (mPFS) was 19.5 months for the Bev group vs. 16.0 months in the CT group (p = 0.150). Patients with a PFI > 12 months showed a mPFS of 20.0 months vs. 15.5 for patients with a PFI < 12 months (p=0.029). Patients with cyclin E1 overexpression showed a mPFS of 15.8 months vs. 19.7 months for those without cyclin E1 overexpression (p=0.05). Benefit of bevacizumab was present only in the subgroup of patients with PFI < 12 months (mPFS 18.6 versus 10.4 months, p=0.002) and in the subgroup of patients with cyclin E1 overexpression (mPFS 16.3 versus 7.0 months, p=0.010). Conclusions:Markers of resistance to chemotherapy such as cyclin E1 overexpression and PFI identify patients with the greatest benefit of bevacizumab. This data could help to discern between maintenance treatment options in the era of PARP inhibitors and anti-angiogenics. Citation Format: Adriana Regina G. Ribeiro, Marcella M. Salvadori, Louise de Brot, Graziele Bovolim, Henrique Mantoan, Felipe Ilelis, Mariana R. Alves, Nayra Soares Amaral, Solange M. Sanches, Joyce Lisboa, Elizabeth S. dos Santos, Ronaldo Pereira, Fabricio S. Castro, Joao Paulo S. Lima, Andrea P. Guimaraes, Glauco Baiocchi, Alexandre Andre B. Da Costa. Cyclin E1 overexpression identifies patients with greater benefit from bevacizumab in platinum sensitive recurrent ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3157.

Tobacco and alcohol are the leading environmental risk factors in the development of human diseases, such as cancer, cardiovascular disease, and liver injury. Despite the copious amount of research on this topic, by 2030, 8.3 million deaths are projected to occur worldwide due to tobacco use. The expression of noncoding RNAs, primarily microRNAs (miRNAs) and long noncoding RNAs (lncRNAs), is modulated by tobacco and alcohol consumption. Drinking alcohol and smoking cigarettes can modulate the expression of miRNAs and lncRNAs through various signaling pathways, such as apoptosis, angiogenesis, and inflammatory pathways—primarily interleukin 6 (IL-6)/signal transducer and activator of transcription 3 (STAT3), which seems to play a major role in the development of diseases associated with these risk factors. Since they may be predictive and prognostic biomarkers, they can be used both as predictors of the response to therapy and as a targeted therapy. Further, circulating miRNAs might be valuable noninvasive tools that can be used to examine diseases that are related to the use of tobacco and alcohol. This review discusses the function of noncoding RNAs in cancer and other human tobacco- and alcohol-associated diseases.

Introdução: O adenocarcinoma é o tipo histológico mais prevalente entre os tumores de pâncreas (90%) e possui alta taxa de mortalidade devido ao seu mau prognostico. A importância de se compreender a carcinogênese pancreática é determinar novos alvos terapêuticos e biomarcadores que auxiliem no diagnostico precoce. A progressão do adenocarcinoma ductal pancreático (ADP) é relacionada ao acúmulo de mutações em lesões pré cancerígenas como a Neoplasia Intraepitelial Pancreática (PanIN). Alterações em oncogenes como: K-ras, EGFR, BRAF, PIK3CA e perda de função em supressores tumorais como CDKN2A e p53 estão envolvidas no desenvolvimento do tumor. Nosso objetivo foi avaliar a frequência de mutações nos genes envolvidos com a progressão do câncer e correlacioná-las com os achados clínicos e a ancestralidade genômica Material e Método: A análise das mutações foi feita por sequenciamento e a ancestralidade através de 40 indels bialélicos em 23 pacientes brasileiros com adenocarcinoma pancreático. Resultados: Dentre todos os genes estudados, as mutações no códon 12 de K-ras foram as mais freqüentes (60%) em nossa amostragem. Em PIK3CA, identificamos uma mutação não descrita (M983K) em um paciente. Mutações nos genes EGFR, BRAF, PIK3CA, CDKN2A e p53 não foram frequentes em nosso estudo. A ancestralidade genômica dos nossos pacientes não diferiu do grupo controle, contudo, observamos que os pacientes com maior componente ancestral africano apresentaram mutações em K-ras. Conclusão: Observamos em nossos pacientes uma menor frequência de mutações em K-ras em relação à maioria dos resultados já descritos, porém mais aproximada à da população coreana, além disto, nossas amostras apresentaram menor freqüência de mutações em CDKN2A e p53 em relação a outras populações já estudadas, reforçando a importância de se levar em consideração diferenças étnicas nos estudos envolvendo alterações genéticas em pacientes com adenocarcinoma pancreático. Devido à alta miscigenação da população brasileira, é interessante que outros estudos como este sejam realizados em outras regiões do país, para caracterizar o perfil genético da população brasileira com adenocarcinoma pancreático. Introduction: Adenocarcinoma is the most prevalent histological type among tumors of the pancreas ( 90 % ) and has high mortality. The importance of understanding the pancreatic carcinogenesis is to determine new therapeutic targets and biomarkers to assist in early diagnosis. The progression of pancreatic ductal adenocarcinoma involved the accumulation of mutations in pre- cancerous lesions, for example, the pancreatic intraepithelial neoplasia ( PanIN ) . Molecular alterations in oncogenes such as K-ras, EGFR , BRAF , PIK3CA and loss function of tumor suppressor, such as CDKN2A and p53 are involved in tumor development. Our objective was to evaluate the frequency of mutations in genes involved in cancer progression and correlate them with the clinical and genomic ancestry Material and Methods: Mutation analysis was made by direct sequencing in 23 samples with pancreatic adenocarcinoma and ancestry was analyzed using 40 biallelic indels. Results: Among all the genes, mutations in codon 12 of K - ras were the most frequent (60 %). In PIK3CA, was identified a new mutation in one patient, do not described in the literature ( M983K ). Mutations in EGFR , BRAF , PIK3CA , CDKN2A and p53 were infrequent in our study . The genomic ancestry of our patients did not differ from the control group, however, we observed that all patients with higher African ancestral component showed mutations in K - ras . Conclusion: We observed in our patients a lower frequency of mutations in K - ras in relation to most of the results already described in other countries, but closer to the Korean population , and our samples showed a lower frequency of mutations in CDKN2A and p53 in relation to other populations, reinforcing the importance of th ethnic differences in studies involving genetic alterations in patients with pancreatic adenocarcinoma . Due to the high miscegenation of the Brazilian population, it is interesting that other studies like this are done in other regions of the country, with the objective of to characterize the genetic profile of Brazilian population with pancreatic adenocarcinoma .

Recently, miR-223-5p was associated with metastasis in HPV negative vulvar samples and in several other tumor types. In the present study, we hypothesized that this microRNA would have a potential impact on in vitro vulvar cancer behavior. We artificially mimicked microRNA expression in vulvar cancer cell line, SW962, derived from lymph node metastasis of vulvar carcinoma, and performed in vitro assays as follows in the result section. Impaired cell proliferation (p Citation Format: Rafael Rocha, Beatriz De Melo Maia, Iara S. Rodrigues, Nayra S. Amaral, Hui Ling, George A. Calin, Fernando A. Soares. Mir-223-5p works as an oncomir in vulvar carcinoma by TP63 regulation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1106.