The BRCA2 p.N372 H i.a.1342A>C Could Regulate the Sensitivity of Ovarian Cancer Cells to Platinum-Based Drugs.

BACKGROUND AND OBJECTIVE: We have previously reported that BRCA2 N372 H i.a.1342A>C heterozygous variation presented in platinum-resistant patients. This study aimed to further investigate the mechanism of BRCA2 N372 H mutation in the development of platinum resistance in ovarian cancer. METHODS: The BRCA2 N372 H i.a.1342A>C was synthesized and used to exchange 1 wildtype allele followed by sequencing to confirm the mutant allele sequence. Plasmids were constructed and transfected into the OVCAR-3 cells after lentiviral packaging. BRCA2 N372 H mRNA was detected by qPCR. BRCA2 protein was assessed by immunoblotting. Binding of the BRCA2 to Rad51 was detected by immunofluorescence staining. Sensitivity of the cells to cisplatin treatment was assessed with CCK-8 assay. RESULTS: It was found that expression of BRCA2 protein in ovarian cancer cells transfected with BRCA2 N372 H i.a.1342A>C gene (2.177 ± 0.003) was significantly increased compared to that of the cells transfected with lenti-EGFP only (1.227 ± 0.003, P < 0.001). Binding of the BRCA2 and Rad51 proteins was significantly increased in the cells with BRCA2 N372 H i.a.1342A>C mutation (3.542 ± 0.24) than that in the cells transfected with lenti-EGFP (1.29 ± 0.32) or empty cells (1.363 ± 0.32, P < 0.001). Cell viability significantly increased in the cells transfected with BRCA2 N372 H mutant gene. The IC50 value was significantly higher in the cells transfected with BRCA2 N372 H mutant gene (1.963 ± 0.04) than that of the cells transfected with lenti-EGFP (0.955 ± 0.03, P < 0.01) or empty cells (1.043 ± 0.007, P < 0.01). CONCLUSION: Over expression of mRNA and protein of BRCA2 was detected in the cells with BRCA2 N372 H i.a.1342A>C mutation but not in the lentivirus negative control (lenti-EGFP) or the cells without transfection (empty cells), which may lead to resistance to platinum-based drugs in ovarian cancer cells through homologous recombination repair pathway.

Association between metabolic syndrome and endometrial cancer risk: a systematic review and meta-analysis of observational studies.

Existing evidence has revealed inconsistent results on the association between metabolic syndrome (MetS) and endometrial cancer (EC) risk. Herein, we aim to better understand this association. Systematic searches of PubMed, EMBASE, and Web of Science through 12 December 2019 were conducted. Observational studies that provided risk estimates of MetS and EC risk were eligible. The quality of the included studies was judged based on the Newcastle-Ottawa scale. Summary odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using a random-effects model. Six studies, comprising 17,772 EC cases and 150,371 participants were included. MetS, diagnosed according to the criteria of the National Cholesterol Education Program-Third Adult Treatment Panel, was associated with an increased risk of EC (OR: 1.62; 95% CI = 1.26-2.07) with substantial heterogeneity (I2 = 78.3%). Furthermore, we found that women with MetS, diagnosed according to the criteria of the International Diabetes Federation, had a significantly higher risk of EC compared to healthy controls (OR: 1.45; 95% CI = 1.16-1.81; I2 = 64.6%). Our findings were generally consistent with the main results in the majority of prespecified subgroups, as well as in sensitivity analyses. In conclusion, MetS is associated with EC risk.

Next-generation sequencing unravels extensive genetic alteration in recurrent ovarian cancer and unique genetic changes in drug-resistant recurrent ovarian cancer.

BACKGROUND: By using a high-throughput sequencing technique, we sought to delineate genetic alterations in recurrent ovarian cancer patients and further compare genetic changes in drug-resistant and -sensitive recurrent ovarian cancer patients. We also sought to study the specificity, sensitivity, and consistency of DNA biomarkers in liquid biopsy specimens and ovarian cancer tissue DNA. METHODS: Tumor tissue specimens and blood samples were obtained from pathologically proven recurrent ovarian cancer patients. Genomic DNA was extracted from tumor tissues, blood cells, ascites, and urine samples. The DNA Library was constructed and sequencing was performed using the Illumina HiSeq 4000 high-throughput sequencing platform. Bioinformatic analysis was done using the Torrent Suite software. RESULTS: Ten patients with pathologically proven drug-resistant recurrent ovarian cancer and 11 patients with sensitive recurrent ovarian cancer were included. The 5-year OS for drug-resistant recurrent ovarian cancer patients (44 ± 11.07 months, 95% CI: 231.24-53.66 months) was significantly lower than that of drug-sensitive recurrent ovarian cancer patients (58 ± 3.97 months; 95% CI: 50.05-65.59 months; p = 0.024) TP53 was the most frequently mutated gene in both drug-resistant (9/10, 90%) and drug-sensitive recurrent ovarian cancers (10/11, 91%). MYC and RB1 had the highest frequency of copy number variations (6/21, 29%) in recurrent ovarian cancers, followed by PIK3CA (3/21, 14%). BRCA2 N372H polymorphism was found in 40% (4/10) of drug-resistant recurrent ovarian cancer patients. The specificity, sensitivity, and consistency of TP53 and BRCA1 in circulating tumor-free DNA and tumor tissue DNA were 100%, 73.7%, 76.2% and 100%, 75%, 95.24%, respectively. CONCLUSION: We uncovered extensive genetic alterations in recurrent ovarian cancer and drug-resistant recurrent ovarian cancer exhibited unique genetic changes compared with recurrent ovarian cancer and drug-sensitive recurrent ovarian cancer. We further showed that high-throughput sequencing using liquid biopsy specimens could provide an effective, specific, and sensitive approach for detecting genetic alterations in ovarian cancer.

Targeted inhibition of the phosphoinositide 3-kinase impairs cell proliferation, survival, and invasion in colon cancer.

BACKGROUND: Colon cancer is the third most common cancer in the world, and its metastasis and drug resistance are challenging for its effective treatment. The PI3K/Akt/mTOR pathway plays a crucial role in the pathogenesis of colon cancer. The aim of this study was to investigate the targeting of PI3K in colon cancer cells HT-29 and HCT-116 in vitro. METHODS: In HT-29 and HCT-116 cells, BEZ235, a dual inhibitor of PI3K/mTOR, and shRNAtarget to PI3KCA were used to inhibit PI3K/Akt/mTOR pathway. The inhibition efficiency of PI3K/Akt/mTOR pathway was detected by RT-PCR and Western blot. Cell proliferation, migration, invasion, and apoptosis were evaluated by Cell Counting Kit-8, Transwell, and flow cytometry assays. The expression of apoptosis-related proteins (cleavage caspase 3, Bcl-2, Bax, and Bim) were also detected. RESULTS: We found that in HT-29 and HCT-116 cells, the treatment of BEZ235 (1 µM) and PI3KCA knockdown inhibited the activation of PI3K/Akt/mTOR pathway and significantly suppressed cell proliferation, migration, and invasion of HT-29 and HCT-116 cells. In addition, we confirmed that knockdown of BEZ235 and PI3KCA induced cell apoptosis through the upregulated levels of cleavage caspase 3 and Bax and downregulated expression of Bcl-2 and Bim. CONCLUSION: Our results indicated that targeted inhibition of the PI3K/Akt/mTOR pathway impaired cell proliferation, survival, and invasion in human colon cancer.

MicroRNA-873 (miRNA-873) inhibits glioblastoma tumorigenesis and metastasis by suppressing the expression of IGF2BP1.

Glioblastoma multiforme (GBM) is known as a highly malignant brain tumor with a poor prognosis, despite intensive research and clinical efforts. In this study, we observed that microRNA-873 (miR-873) was expressed at low levels in GBM and that the overexpression of miR-873 dramatically reduced the cell proliferation, migration, and invasion of GBM cells. Our further investigations of the inhibition mechanism indicated that miR-873 negatively affected the carcinogenesis and metastasis of GBM by down-regulating the expression of IGF2BP1, which stabilizes the mRNA transcripts of its target genes. These results demonstrate that miR-873 may constitute a potential target for GBM therapy.

[Integrated evaluation on system reliability of a space metabolism simulation device].

OBJECTIVE: To define the lower confidence limit of the system reliability of the space metabolism simulation device by using the test data of the comprising units in their development phases. METHOD: A new method for defining the environmental conversion factors of failure times has been given basing on the AMSAA model, and the MTBF point evaluated values and lower confidence limits of the units were defined, at last, the lower confidence limit of the system reliability was obtained using L-M method. RESULT: The lower confidence limit of the system reliability under confidence level of 0.8 is 0.9281, which is consistent with the result obtained by the classic system reliability evaluation method. CONCLUSION: Integrated system reliability evaluation using the proposed method can well satisfy the need in engineering.