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The success of cloning by somatic cell nuclear transfer depends on the efficiency of nuclear reprogramming, with the cycle stage of the donor cell playing a crucial role. Therefore, the aim was to evaluate three different approaches for cell cycle synchronization: (i) serum starvation (SS) for 1 to 4 days, (ii) contact inhibition (CI) for 1 to 3 days, and (iii) using cell cycle regulatory inhibitors (dimethyl sulfoxide, cycloheximide, cytochalasin B, or 6-dimethylaminopurine) for 1 and 2 days, in terms of their effects on synchronization in G0/G1 phases and viability of collared peccary skin fibroblasts. Flow cytometry analysis revealed that SS for 4 days (79.0% ± 1.6) and CI for 3 days (78.0% ± 1.4) increased the percentage of fibroblasts in G0/G1 compared to growing cells GC (68.1% ± 8.6). However, SS for 3 and 4 days reduced the viability evaluated by differential staining (81.4% ± 0.03 and 81.6% ± 0.06) compared to growing cells (GC, 95.9% ± 0.06). CI did not affect the viability at any of the analyzed time intervals. No cell cycle inhibitors promoted synchronization in G0/G1. These results indicate that CI for 3 days was the most efficient method for cell cycle synchronization in peccary fibroblasts.

INTRODUCTION: PMS1 is part of the cluster of genes related to Mismatch repair genes (MMR) and has been used for investigation in oncogenetic panels. The MMR genes play an important role in tumor control and progression, however, the role of PMS1 in this process still poorly understood and information about its role in increasing the risk of developing a hereditary cancer predisposition syndrome (SHPC) is not completely understood.AIM: To characterize clinically and genetically cancer patients with NCCN criteria for SHPC and carriers of variants in the PMS1 gene.METHODOLOGY: A total of 368 patients suspected of having SHPC, according to the National Comprehensive Cancer Network (NCCN) criteria, were investigated using a Next-Generation Sequencing (NGS) in a panel containing 31 genes. Those that showed variation in the PMS1 gene were grouped, and the tumors were characterized in clinical and molecular aspects.RESULTS: From the 368 patients analyzed, 6.8% (25/368) patients presented Variants of Uncertain Significance (VUS) in PMS1. There was a case of Breast Cancer (BC) with a variant in PMS1, not described in ClinVar, presented in heterozygosis, probably pathogenic [Chr2:190.738,325 NM_000534: c.2578delA: p.(Arg860.Glufs*14)]. Besides, this case was associated with a family history (FH) of cancer breast cancer and melanoma. It is worth mentioning that only one patient was a man, with colorectal cancer (CRC), and his FH also was CRC in addition to stomach cancer. The only change founded was the presence of VUS NM_000534:c.1615A>G:p.(Met539Val) which was identified in 32% (8/25). This VUS was the only genetic alteration observed in 20% (5/25) patients with tumors in the breast (3), thyroid (1), CRC (1), and ovary (1). One of these patients had bilateral breast cancer and thyroid tumor. The patients had their age at diagnosis ranged from 25 to 65 years, with an average of 41 years. The carriers of variants exclusively in PMS1 [40% (10/25)] had different tumors: breast (6); CRC (3); ovary (2); lymphoma (1); thyroid (1); others 32% (n = 8/25) that presented additional VUS in different genes: [ATM (2); BARD (1); CDH1 (1); CHECK2 (1); NBN (1); APC (1); POLE (1)]. Different pathogenic variants (PV) [28% (n = 7/25)] also were identified, besides the previous PMS1 VUS described. The PV was founded in ATM (1); BRCA1 (3); NF1 (1); TP53 (1); MUTYH-heterozygosis (1), and PMS1 (1) gene. A double pathogenic variation was identified in a patient (MUTYH in heterozygosis and BRCA1). The findings in PMS1, even currently classified as VUS, should highlight observation for the clinical and familiar history. Genetic panels should be increasingly used in the investigation of SPHC.CONCLUSION: The roles of PMS1 in cancer progression need further investigations. A few numbers of reports have been identified on germline PMS1 mutations, considering defined disease phenotypes. Therefore, the performance of genetic panels, including the PMS1 gene, in SHPC further investigations will expand our knowledge and permit precise genetic counseling. Citation Format: Isabelle JoyceLima Silva-Fernandes, Clarissa Gondim Picanço-Albuquerque, Maria Claudia dos Santos Luciano, Deysi Viviana Tenazoa Wong, Maria Júlia Barbosa Bezerra, Flavio Silveira Bitencourt, Francisca Fernanda Barbosa Oliveira, Paulo Goberlanio de Barros Silva, Rosane Oliveira Sant´anna, Marcos Venicio Alves Lima. Pms1 gene: A new risk-mutation description? [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS16-30.

INTRODUCTION: HBOC syndrome is directly associated with a variant in BRCA1/2 genes and the diagnosis may increase anxiety and depression levels because of the need for preventive measures. Psychic impacts on women with mutations have been further explored in the literature, although the emotional aspects linked to the diagnosis in male breast cancer is poorly known. AIM: This work aimed to compare the anxiety, depression, and quality of life levels in male and female cancer patients HBOC-suspected, immediately after the result of the genetic test. METHODOLOGY: A retrospective case-control study was carried out with five men matched for age with 39 women, both with National Comprehensive Cancer Network (NCCN) criteria for HBOC. The Hospital Anxiety and Depression (HAD) scale and the WHOQOL-bref quality of life inventory were used. Sociodemographic, clinical, molecular, and psychological data were analyzed using χ2/Fisher's exact and Mann-Whitney, Friedman/Dunn, and Wilcoxon tests (SPSS, 20.0; p 45 years old, all recruited women had breast cancer as the primary tumor and eight of them (20.5%) had ovary cancer. We had two male breast cancer and three prostate cancer Gleason>7. Sociodemographic characteristics did not differ between the genders (p> 0.05). Men had a higher prevalence of pathogenic mutation (p=0,031). Women had a higher prevalence of BRCA1 mutations (p=0,014) while men had of BRCA2 mutation (p=0,001). There was no difference in the anxiety and depression levels immediately after receiving the result, but it was observed that only women had lower quality of life in the physical domain (p=0,048). This domain showed the lowest scores in female patients (p Citation Format: Francisca Fernanda Barbosa Oliveira, Paulo Goberlanio de Barros Silva, Isabelle JoyceLima Silva-Fernandes, Clarissa Gondim Picanço-Albuquerque, Maria Claudia dos Santos Luciano, Maria Júlia Barbosa Bezerra, Rosane Oliveira Sant´anna, Flavio Silveira Bitencourt, Marcos Venicio Alves Lima. Hereditary breast and ovarian cancer syndrome (HBOC) diagnosis impacts more the life quality of women than men: A case-control report [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS9-37.

INTRODUCTION: The male breast cancer (MBC) is a rare disease, responsible for about 0.2% of all cancers and 0.1% of deaths for male cancer. Despite the rarity of the disease, statistics indicate that the incidence of MBC has been increasing significantly. The main risk factors for the development of MBC include old age, hormonal imbalance, radiation exposure, and family history of breast cancer. Hereditary breast and ovarian cancer (HBOC) is a syndrome associated with mutations in the BRCA1 and BRCA2 genes that have been related to MBC that as a rare disease, still poorly understood. METHODOLOGY: Family history analyzes, type and tumor staging, molecular markers expression, hormone receptors (RE, RP, and HER2), and cell proliferation marker (KI67) analysis were performed by immunohistochemistry. The presence of genetic mutations and the frequency of these mutations were evaluated in 31 genes of HBOC suspected patients treated at the Haroldo Juaçaba Hospital between the years 2009 to 2020. RESULTS: A total of 236 patients were diagnosed with breast cancer and submitted to a genetic test. Six of them were male patients aged between 54 and 73 years. His family history indicated that 83.3% had first-degree relatives affected by breast cancer. Different pathological staging was founded after tumors evaluation with the presence of micrometastases (P3) and higher levels of invasion in the lymph nodes. The expression of the cell proliferation marker (KI67) indicated low levels of expression except for patient P5. All patients presented expression of two hormone receptors (ER and RP) and no expression for HER2, except for one patient (P1), who had bilateral breast cancer, with HER2 expression (+2) on the right breast (first tumor) and no RP and HER2 expression in the left breast (second tumor), besides a moderate expression of KI67 (40%) (Table 1). The oncogenetic evaluation indicated that 83.33% of the patients had mutations described in the clinical database (ClinVar) 60% of them were pathogenic. The variants founded were in the PALB2 (16.66%), BRCA1 (33.33%), and BRCA2 (33.33%) genes (Table 1). The pathogenic mutations founded were located in the BRCA 1 and BRCA2 genes. One patient (P5) did not present mutations or VUS. The patient without the mutation (P5) presented a tumor with T3N1 pathological staging, low expression of estrogen (10%) and progesterone (10%) receptors, absence of HER2 expression, and high capacity of proliferation, indicated by the expression of KI67 (80%). The results indicate heterogeneity in the histological and molecular patterns of the patients evaluated, besides the oncogenetic patterns that may be associated with HBOC, especially those whose variants founded were pathogenic.CONCLUSION: Due to the low frequency of male breast cancer, the oncological data of these patients are relevant for epidemiological, pathological, and oncogenetic characterization, improving the characterization and identification of patterns for this pathology.Research Sponsor: Ministerio da Saude, Brasil (PRONON) Table 1. Histopathologic molecular markers and variation gene characterization of male breast cancer evaluated in Hospital Haroldo Juaçaba, Northest of Brazil, between 2009 and 2020. *VUS #Pathogenic variation.AgeFamiliar historyMolecular markersVariationPatientDiagnosticLast clinical evaluationBCOtherSubtype tumorPathological stagingRERPHER2KI67GeneSpecificityP14857YesPancreas; CCPLUMINAL A(R) pT1cpN1a(L) pT1B(R) 80%(L) 80%(R) 20%(L) negative(R) +2(L) negative(R) 18%(L) 40%PALB2*c. 3257G>A:Arg1086GlnP27274YesNoLUMINAL B(L)pT2 pN3a90%100%Negative20%BRCA1*c. 754C>T:Arg252CysP36466NoNoLUMINAL ApT2pN1mic90%70%Negative10%BRCA1#c. 5266dupC: p.Gln1756Pro fs74P45457YesNoLUMINAL A(L)T4bN290%90%Negative10%BRCA2#c. 4808delA: pAsn1603Thr fs14P55858YesProstaticLUMINAL A(R) T3N110%10%Negative80%Not detectedNot detectedP65858YesProstaticLUMINAL B(R) T4dN060%60%Negative30%BRCA2#c. 4808delA: pAsn1603Thr fs14 Citation Format: Maria Claudia dos Santos Luciano, Maria Júlia Barbosa Bezerra, Isabelle JoyceLima Silva-Fernandes, Paulo Goberlanio de Barros Silva, Clarissa Gondim Picanço-Albuquerque, Rosane Oliveira Sant´anna, Francisca Fernanda Barbosa Oliveira, Flavio Silveira Bitencourt, Marcos Venicio Alves Lima. Oncogenetic caracterization and imunohistochemistry patterns of male cancer breast diagnosticated in Haroldo JuaÇaba hospital, Northeast of Brazil [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS14-27.

The imprinted H19 long non-coding RNA, a knowing oncofetal gene, presents a controversial role during the carcinogenesis process since its tumor suppressor or oncogenic activity is not completely elucidated. Since H19 lncRNA is involved in many biological pathways related to tumorigenesis, we sought to develop a non-cancer lineage with CRISPR-Cas9-mediated H19 knockdown (H19-) and observe the changes in a cellular context. To edit the promoter region of H19, two RNA guides were designed, and the murine C2C12 myoblast cells were transfected. H19 deletion was determined by DNA sequencing and gene expression by qPCR. We observed a small deletion (~ 60 bp) in the promoter region that presented four predicted transcription binding sites. The deletion reduced H19 expression (30%) and resulted in increased proliferative activity, altered morphological patterns including cell size and intracellular granularity, without changes in viability. The increased proliferation rate in the H19- cell seems to facilitate chromosomal abnormalities. The H19- myoblast presented characteristics similar to cancer cells, therefore the H19 lncRNA may be an important gene during the initiation of the tumorigenic process. Due to CRISPR/Cas9 permanent edition, the C2C12 H19- knockdown cells allows functional studies of H19 roles in tumorigenesis, prognosis, metastases, as well as drug resistance and targeted therapy.

Natural pterocarpans and synthetic 5-carba-pterocarpans are isosteres in which the oxygen atom at position 5 in the pyran-ring of pterocarpans is replaced by a methylene group. These 5-carba-analogues were obtained in good yields through the palladium-catalyzed oxyarylation of alcoxy-1,2-dihydronaphthalens with o-iodophenols in PEG-400. They were evaluated on human cancer cell lineages derived respectively from prostate tumor (PC3, IC50 = 11.84 μmol L−1, SI > 12)) and acute myeloid leukemia (HL-60, IC50 = 8.81 μmol L−1, SI > 16), highly incident cancer types presenting resistance against traditional chemotherapeutics. Compound 6c (LQB-492) was the most potent (IC50 = 3.85 μmol L−1, SI > 37) in SF-295 cell lineage (glioblastoma). Such findings suggest that 5-carba-pterocarpan can potentially be new hit compounds for further development of novel antiproliferative agents.

The study aimed to evaluate cytotoxicity and antitumor activity of perillaldehyde 1,2-epoxide (PE), a p-menthane monoterpene derivative against four human tumor cell lines ovarian cancer (OVCAR-8), colon carcinoma (HCT-116), glioblastoma (SF-295) and leukemia (HL-60) using the colorimetric MTT assay. PE showed a high degree of inhibition of cell proliferation (GI = 95.66 to 99.71%) and IC50 16.14 μM (± 1.86), 23.61 μM (± 1.13), 21.99 μM (± 2.64) and, 9.70 μM (± 1.01) against tumor cells, respectively. Then, in vivo antitumor activity of the PE was assessed in sarcoma 180-bearing mice. Tumor growth inhibition rates were 33.4, 56.4 and 66.6% at doses of 100 and 200 mg/kg/day for the PE and 25 mg/kg/day for 5-FU intraperitoneal treatments, respectively. Toxicological effects related to the spleen, kidneys, liver, and hematological were investigated in mice submitted to treatment. Furthermore, histopathological analyses of these organs were absent of any morphological changes in the animals treated with PE. The viability of HL-60 cells was affected by perillaldehyde 1, 2-epoxide after an exposure period of 72 h when analyzed by trypan blue exclusion. PE reduced the number of viable cells associated with an increase in non-viable cells, which contributes to the increased number of dead cells in the morphological analysis. The incorporation of ethidium bromide/acridine orange, the treated cells suggests cytotoxicity via apoptosis and necrosis. So on the results, we conclude that PE presents cytotoxic and antitumoral activity through apoptotic and necrotic processes. Key words: Essential oils, p-menthanes, natural products, cytotoxicity, antitumor activity, sarcoma 180.

Intestinal mucositis is a common complication associated with 5-fluorouracil (5-FU), a chemotherapeutic agent used for cancer treatment. Troxerutin (TRX), a semi-synthetic flavonoid extracted from Dimorphandra gardneriana, has been reported as a potent antioxidant and anti-inflammatory agent. In the present study, we aimed to evaluate the effect of TRX on 5-FU-induced intestinal mucositis. Swiss mice were randomly divided into seven groups: Saline, 5-FU, TRX-50, TRX-100, TRX-150, Celecoxib (CLX), and CLX + TRX-100. The weight of mice was measured daily. After treatment, the animals were euthanized and segments of the small intestine were collected to evaluate histopathological alterations (morphometric analysis), levels of malondialdehyde (MDA), myeloperoxidase (MPO), glutathione (GSH), mast and goblet cell counts, immunohistochemical analysis, and cyclooxygenase-2 (COX-2) activity. Compared to the saline treatment, the 5-FU treatment induced intense weight loss and reduction in villus height. TRX treatment (100 mg/kg) prevented the 5-FU-induced histopathological changes and decreased oxidative stress by decreasing the MDA levels and increasing GSH concentration. TRX attenuated inflammatory process by decreasing MPO activity, intestinal mastocytosis, and COX-2 expression. TRX also reversed the depletion of goblet cells. Our findings suggest that TRX at a concentration of 100 mg/kg had chemopreventive effects on 5-FU-induced intestinal mucositis via COX-2 pathway.

Pistia stratiotes trata-se de uma macrofita aquatica flutuante que se desenvolve livremente no espelho d’agua, e conhecida como alface-d’-agua, erva-de-santa-luzia, repolho-d’agua e golfo. Objetivou-se com este trabalho avaliar como a especie P. stratiotes esta sendo utilizada no desenvolvimento cientifico e tecnologico, atraves do perfil quantitativo das publicacoes e deposito de patentes com o genero Pistia e a especie P. stratiotes. A prospeccao foi realizada com base em periodicos depositados no Web of Science e base de dados de patentes, European Patent Office (ESPACENET), World Intelectual Property Organization (WIPO), United States Patent and Trademark Office (USPTO) e do Instituto Nacional de Propriedade Industrial (INPI) do Brasil. Os resultados demonstram que o EUA foi o pais que mais publicou pesquisas com a especie, o Brasil ficou em terceiro lugar no numero de trabalhos publicados, atras de paises como EUA e India. Os dados levantados por patentes resultaram num total de 116 documentos para o termo Pistia e 70 documentos para o termo Pistia stratiotes, considerando a somatoria de todos os documentos encontrados em todas as bases de dados buscadas.

Growing evidence suggests that aberrant epigenetic regulation of gene function is strongly related to the genesis of cancer. Unlike genetic mutations, the ability to reprogram the epigenetic landscape in the cancer epigenome is one of the most promising target therapies in both treatment and reversibility of drug resistance. Epigenetic alterations in cancer development and progression may be the basis for the individual variation in drug response. Thus, this review focuses on the emerging area of pharmaco(epi)genomics, specifically highlighting epigenetic reprogramming during tumorigenesis and how epigenetic markers are targeted as a therapy (epidrugs) and the clinical implications of this for cancer treatment.

This work aimed to evaluate the cytotoxic potential against cancer cells of Passiflora genus plant species cultivated in Brazil and identify the mechanism of cytotoxicity induced by the most promising extract. Ethanolic extracts from the leaves of 14 Passiflora species were obtained by accelerated solvent extraction and in vitro cytotoxicity evaluated against cancer cell lines using MTT assay at a single concentration of 50 μg/ml. Additionally, the IC50 of the Passiflora alata (ELPA) leaf extracts was determined against both cancer (HCT-116, SF-295, OVACAR-8, and HL-60), and non-cancer cells (PBMC). The ELPA flavonoids were identified by HPLC-DAD and UHPLC-MS/MS. The morphological analyses, using light and fluorescence microscopy, and cell cycle and DNA fragmentation analysis, using flow cytometry, were evaluated to study the mechanism of cell death induced by ELPA in HL-60 cells. Among the Passiflora leaf extracts evaluated; ELPA stood out with high cytotoxic activity, followed by Passiflora capsularis and Passiflora quadrangularis with varying high and low cytotoxic activity. ELPA presented high cytotoxic potency in HL-60 (IC50 19.37 μg/ml), and without cytotoxicity against PBMC, suggesting selectivity for cancer cells. The cytotoxic activity of ELPA may well be linked to the presence of ten identified flavonoids. Cells treated with ELPA presented the hallmarks typical of apoptosis and necrosis, with cell cycle arrest in the G2/M phase. From among the studied species, ELPA presented greater cytotoxic activity, possibly a consequence of synergistic flavonoid action, which induces cell death by apoptosis and necrosis. Key words: Cancer, cytotoxicity, ethanolic extract, Passiflora, Passiflora alata.

Natural α,β-amyrins were isolated from endemic Brazilian Esenbeckia grandiflora Mart., and eight synthetic derivatives were obtained by esterification reactions with bromo acetate, followed by amine treatment. The structures of the all compounds were confirmed by 1H and 13C nuclear magnetic resonance (NMR), Fourier transform infrared (FTIR) and high-resolution mass spectrometry (HRMS) data analysis. The derivatives were screened for cytotoxic activity against human tumor cell-lines PC3 (prostate carcinoma), HCT-116 (colon carcinoma) and HL60 (leukemia). HCT-116 and PC3 cell-lines showed weak tumor growth inhibition (range of 13.9-25.4 and 10.3-28.8%, respectively), but the derivatives presented moderate activity against HL60 (range of 13.6-59.0%). Diethyl, aniline, morpholine and imidazole moieties presented higher activities (range of 45.9-59.0%).