Breast cancer gene 1 (BRCA1) and BRCA2 are tumor suppressors involved in DNA damage response and repair. Carriers of germline pathogenic or likely pathogenic variants in BRCA1 or BRCA2 have significantly increased lifetime risks of breast cancer, ovarian cancer, and other cancer types; this phenomenon is known as hereditary breast and ovarian cancer (HBOC) syndrome. Accurate interpretation of BRCA1 and BRCA2 variants is important not only for disease management in patients, but also for determining preventative measures for their families. BRCA1:c.132C>T (p.Cys44=) is a synonymous variant recorded in the ClinVar database with "conflicting interpretations of its pathogenicity". Here, we report our clinical tests in which we identified this variant in two unrelated patients, both of whom developed breast cancer at an early age with ovarian presentation a few years later and had a family history of relevant cancers. Minigene assay showed that this change caused a four-nucleotide loss at the end of exon 3, resulting in a truncated p.Cys44Tyrfs*5 protein. Reverse transcription-polymerase chain reaction identified two fragments (123 and 119 bp) using RNA isolated from patient blood samples, in consistency with the results of the minigene assay. Collectively, we classified BRCA1:c.132C>T (p.Cys44=) as a pathogenic variant, as evidenced by functional studies, RNA analysis, and the patients' family histories. By analyzing variants recorded in the BRCA Exchange database, we found synonymous changes at the ends of exons could potentially influence splicing; meanwhile, current in silico tools could not predict splicing changes efficiently if the variants were in the middle of an exon, or in the deep intron region. Future studies should attempt to identify variants that influence gene expression and post-transcription modifications to improve our understanding of BRCA1 and BRCA2, as well as their related cancers.

BACKGROUND Recently, the upregulation of LINC00963 expression has been reported in various cancer subtypes. LINC00963 expression can promote cancer cell invasion and metastasis. However, the clinical significance of LINC00963 in cervical and endocervical cancer (CESC) has remained relatively unexamined. MATERIAL AND METHODS We assessed the mRNA expression of LINC00963 in patients with CESC based on data acquired from The Cancer Genome Atlas (TCGA) to determine pathways involved in CESC pathogenesis with respect to LINC00963. We included 3 normal and 304 tumor samples in this study. RESULTS The scatter plot and paired plot showed differences in LINC00963 expression between normal and tumor samples (P<0.01). Overall survival (OS) analysis revealed that CESC patients with high expression of LINC00963 demonstrated worse prognosis than CESC patients with low expression of LINC00963 (P<0.01). Multivariate analysis with the Cox proportional hazards model indicated that the expression of LINC00963 (HR 0.297; 95% CI 0.115-0.776; P=0.012) and primary therapy outcome (HR 0.162; 95% CI 0.059-0.446; P=0.001) were independent prognostic factors for patients with CESC. GSEA results showed that reactome biological oxidations, inflammasomes, apoptosis, toll-like receptor signaling pathway, JAK/STAT signaling pathway, and NF-kappaB activation were differentially enriched in CESC samples with the high LINC00963 expression phenotype. CONCLUSIONS Our results confirmed the association of significantly high levels of LINC00963 expression in CESC with several observed clinical features. LINC00963 may be a potentially useful prognostic molecular biomarker associated with poor survival in patients with CESC.

Predisposition of germline BRCA1/2 mutations ( gBRCA MUT ) increases the risk of breast and ovarian cancer in females, but the mutation prevalence and spectrum are highly ethnicity-specific with different recurrent mutations being reported in different populations. Hereby, we performed hybridization-based target sequencing of BRCA1/2 in 530 ovarian cancer patients from Henan, the central region of China, followed by haplotype analysis of six short tandem repeat (STR) markers in the patients with recurrent mutations to determine their founder effect. About 28.3% (150/530) of the OC patients in our cohort harbored gBRCA MUT ; of the 151 mutations, 117 in BRCA1 and 34 in BRCA2 , identified in this study, BRCA1 :c.5470_5477del, c.981_982del, and c.4065_4068del are the top three mutants, recurrently detected in eight, seven, and six independent patients respectively. Haplotype analysis identified a region of 0.6 MB genomic length covering BRCA1 highly conserved across all eight carriers of BRCA1 :c.5470_5477del, but not c.981_982del, suggesting a consequence of founder effect. Retrospective analysis in a subgroup of serous ovarian cancer patients revealed gBRCA MUT status was not associated with the progression-free survival (PFS); instead, an expression of Ki-67% ≥50% was associated with a shorter PFS ( p = 0.041). In conclusion, patients with pathogenic or likely pathogenic gBRCA MUT account for 28.3% of the OC cases from Henan, and BRCA1 :c.5470_5477del, the most frequently detected mutation in Henan patients, is a founder mutation in the population.

Preeclampsia (PE) is a pregnancy-specific syndrome that has severe implications on perinatal mortality and morbidity. Excessive apoptosis of trophoblasts induced by hypoxia may be associated with the development of PE, but the exact pathogenesis is unknown. Forkhead box O transcription factor 3a (FOXO3a) is activated under hypoxic conditions. Furthermore, hypoxia-inducible factor-1 alpha (HIF-1 alpha) is sensitive to variations in partial oxygen pressure. Thus, the aims of the present study were to investigate the expression levels of HIF-1 alpha and FOXO3a in placental samples of early onset severe PE, and their effect on trophoblastic apoptosis under hypoxic conditions. Cobalt chloride was used to establish the hypoxic model. The present study examined the expression levels of HIF-1 alpha and FOXO3a in the placental tissues and HTR8/SVneo cells under hypoxic conditions. It was found that HIF-1 alpha and FOXO3a were highly expressed in placental tissues of patients with PE and in HTR8/SVneo cells under hypoxic conditions. Furthermore, knockdown of FOXO3a using a specific small interfering RNA (siRNA) decreased apoptosis in HTR8/SVneo cells. Moreover, it was found that after knockdown of HIF-1 alpha using siRNA, FOXO3a expression and the apoptotic rate were reduced in HTR8/SVneo cells. Therefore, the present results indicated that the elevated expression of HIF-1 alpha increased trophoblastic apoptosis by regulating FOXO3a, which may be involved in the pathogenesis of PE.

INTRODUCTION:Pre-eclampsia (PE) is a serious complication of pregnancy, and the likely pathogenic basis of early onset PE are placental dysfunction and increased oxidative stress. Resveratrol (RES) is a potent antioxidant which has shown beneficial effects in many diseases. The aim of this study was to investigate the protective effects of RES against oxidative stress-induced damage in trophoblasts, and elucidate the potential mechanisms.METHODS:We established an in vitro model of oxidative stress by exposing the human first-trimester extravillous trophoblast cell line HTR8/SVneo to H2O2. The level of oxidative stress was reflected by ROS, MDA and SOD. The viability of cells was determined by the MTS assay. Apoptosis was detected using Annexin V-FITC staining and flow cytometry. Levels of SIRT1(sirtuin 1) and autophagy-related proteins (LC3, Beclin-1, p62) were detected by western blot. Autophagosomes were observed by transmission electron microscopy (TEM).RESULTS:Pre-treatment with RES significantly ameliorated H2O2-induced cytotoxicity, morphological damage, oxidative stress and apoptosis. Mechanistically, RES restored the levels of SIRT1 and autophagy-related proteins including LC3-II, Beclin-1 and p62 that were dysregulated by H2O2. Blocking autophagy by 3-methyladenine (3-MA) completely abolished the protective effects of RES, as did knocking down SIRT1.CONCLUSION:RES may protect human trophoblasts against H2O2-induced oxidative stress by activating SIRT1-dependent autophagy, and therefore has therapeutic potential in PE.

Breast cancer (BC) is the most frequently diagnosed tumor in women worldwide. Although the combination of surgery and Taxol chemotherapy can achieve a certain therapeutic effect, patients often develop drug-resistance, resulting in a poor prognosis. Therefore, it is significative to seek the molecular mechanism of chemotherapy resistance. Recent studies have found that abnormal epigenetic regulation in breast cells changes the expression of key genes, which can lead to the occurrence, development, and maintenance of cancer, even related to the development of drug-resistance. Therefore, in this study, we performed methylated DNA immunoprecipitation-sequencing (MeDIP-seq) to reveal the difference in methylation between breast cancer drug-resistant cells and sensitive cells. A total of 55076 differentially methylated genes (DMGs) were detected, including 21061 hypermethylated DMGs and 34015 hypomethylated DMGs. Moreover, Gene Ontology (GO) analysis and KEGG pathway analysis reveal the function and pathway of screening genes. These results indicate that DNA methylation may be involved in regulating the occurrence and development of breast cancer.

Background. Preeclampsia (PE) is a pregnancy-specific hypertension syndrome and is one of the leading causes of maternal and perinatal morbidity and mortality. Long noncoding RNAs (lncRNAs) have been reported to be abnormally expressed in many diseases, including preeclampsia. The present study is aimed at identifying the key genes and lncRNA-associated competing endogenous RNA (ceRNA) networks in early-onset preeclampsia (EOPE).Methods.We investigated expression profiles of differentially expressed lncRNAs (DElncRNAs) and genes (DEGs) in placental tissues of EOPE and healthy controls with Human LncRNA Array v4. The potential functions of DEGs and DElncRNAs were predicted using the clusterProfiler package. The lncRNA-mRNA coexpression network was constructed via Pearson's correlation coefficient. The protein-protein interaction (PPI) network of DEGs was constructed, and the hub genes were obtained using the STRING database and Cytoscape. The ceRNA networks were constructed based on miRWalk and LncBase v2. qRT-PCR was performed to confirm the expression of lncRNA MIR193BHG, PROX1-AS1, and GATA3-AS1. ROC curves were performed to assess the clinical value of lncRNA MIR193BHG, PROX1-AS1, and GATA3-AS1 in the diagnosis of EOPE.Results.We found 6 hub genes (SPP1, CCR2, KIT, ENG, ACKR1, and FLT1) altered in placental tissues of EOPE and constructed a ceRNA network, including 21 lncRNAs, 3 mRNAs, and 69 miRNAs. The expression of lncRNA MIR193BHG and GATA3-AS1 were elevated and showed good clinical values for diagnosing EOPE.Conclusion.This study provides novel insights into the lncRNA-related ceRNA network in EOPE and identified two lncRNAs as potential prognostic biomarkers in EOPE.

OBJECTIVE:The purpose of this study was to investigate the expression of atypical chemokine receptor 2 (ACKR2, D6) in different types of preeclampsia (PE) and its effects on trophoblast proliferation and apoptosis.METHODS:The subjects were divided into four groups: early-onset PE group (EOPE, n = 30), late-onset PE group (LOPE, n = 30), preterm birth group (PB, n = 30), and normal group (N, n = 30). The expression of ACKR2 in placentas was evaluated using immunohistochemistry, qRT-PCR, and Western blot. The trophoblast cell line JAR was cultured to detect the expression of ACKR2 after simulating hypoxic conditions with cobalt chloride (CoCl2). The effects on cell proliferation, apoptosis, and expression of the chemokine CCL2 were analyzed after silencing ACKR2 with siRNA.RESULTS:ACKR2 was decreased in placentas of EOPE and PB groups at the protein and mRNA level，compared to the normal group. No statistical differences were found between EOPE and PB groups, or between LOPE and normal groups. In our in vitro work, we found that the expression of ACKR2 decreased after treatment with 150 μmol/L, 200 μmol/L, and 250 μmol/L of CoCl2. After ACKR2 was silenced, the degree of cellular proliferation decreased, while apoptosis and CCL2 expression increased.CONCLUSION:The changes of ACKR2 expression in placentas of PE may be related to gestational weeks. Hypoxia inhibits the expression of ACKR2 in placentas. Abnormal expression of ACKR2 in PE may lead to dysfunction of trophoblast, and ACKR2 is an essential player in the immunoregulation of the placental chemokine CCL2.

Objective To investigate the role of microRNA-26a (miR-26a) in cell migration and invasion in cervical cancer and its regulatory effects on high mobility group protein A1(HMGA1). Methods Both HeLa and SiHa cells were divided into four groups:miR-26a mimic group,mimic control group,miR-26a inhibitor group and inhibitor control group. MiR-26a expression was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Target genes for miR-26a-5p were predicted by bioinformatics and verified by dual-luciferase reporting system. HMGA1 expression was evaluated by Western blot and qRT-PCR. Cell migration and invasion were analyzed by in vitro assays. Results (1) HMGA1 was predicted as one of the potential targets of miR-26a by TargetScan. Results of dual-luciferase activity assay further con-firmed that HMGA1 was directly regulated by miR-26a. (2) Expression of miR-26a and HMGA1 at mRNA level was respectively enhanced and inhibited in miR-26a mimic-transfected HeLa as well as SiHa cells as compared with those in the corresponding mimic control groups (P<0.05). On the contrary, expression of miR-26a and HMGA1 at mRNA level was respectively reduced and increased in miR-26a inhibitor groups as compared with those in the corresponding inhibitor control groups(P<0.05). (3) Expression of HMGA1 in miR-26a mimic-transfected HeLa and SiHa cells was lower than that in the corresponding mimic control groups(P<0.05). But expression of HMGA1 in miR-26a inhibitor groups was higher than that in the corre-sponding inhibitor control groups(P<0.05). (4) Abilities of cell migration and invasion were suppressed in miR-26a mimic groups as compared with those in the corresponding mimic control groups,but were enhanced in miR-26a inhibitor groups as compared with those in the corresponding inhibitor control groups(P<0.05). Conclusion MiR-26a can inhibit the proliferation and invasion of HeLa and SiHa cells through targeting HMGA1,suggesting that miR-26a is closely related to cervical cancer and might be associated with chemo-therapy resistance in cervical cancer. This study might provide a new strategy for the prevention and treat-ment of cervical cancer.

Objective. In search of novel mechanisms leading to the development of gastric cancer, we analyzed differences of protein expression pattern between gastric cancer cell lines of MGC80-3, BGC-823, SGC-7901, AGS and normal gastric epithelial cell line GES-1. Methods. Cells were cultured in vitro and analyzed by proteomics approach. Results. Twenty-two different expression proteins were detected and identified. Nine of them were up-regulated, 10 of them were down-expressed in gastric cancer cells, and 3 of them only expressed in normal gastric epithelial cell. Conclusion. The results suggest that expressed proteins are different between gastric cancer cells and normal cells. These identified proteins may be important in gastric cancer formation and represent potential targets for tumor markers. [Life Science Journal. 2008; 5(4): 28-32] (ISSN: 1097-8135).