OBJECTIVE: We aimed to evaluate the prognostic and predictive value of the nucleotide excision repair-related gene GTF2H5, which is localized at the 6q24.2-26 deletion previously reported by our group to predict longer survival of high-grade serous ovarian cancer patients. METHODS: In order to test if protein levels of GTF2H5 are associated with patients' outcome, we performed GTF2H5 immunohistochemical staining in 139 high-grade serous ovarian carcinomas included in tissue microarrays. Upon stratification of cases into high- and low-GTF2H5 staining categories (> and < or = median staining, respectively) Kaplan-Meier and log-rank test were used to estimate patients' survival and assess statistical differences. We also evaluated the association of GTF2H5 with survival at the transcriptional level by using the on-line Kaplan-Meier plotter tool, which includes gene expression and survival data of 855 high-grade serous ovarian cancer patients from 13 different datasets. Finally, we determined whether stable short hairpin RNA-mediated GTF2H5 downregulation modulates cisplatin sensitivity in the SKOV3 and COV504 cell lines by using cytotoxicity assays. RESULTS: Low expression of GTF2H5 was associated with longer 5-year survival of patients at the protein (hazard ratio [HR], 0.52; 95% CI, 0.29 to 0.93; p=0.024) and transcriptional level (HR, 0.80; 95% CI, 0.65 to 0.97; p=0.023) in high-grade serous ovarian cancer patients. We confirmed the association with 5-year overall survival (HR, 0.55; 95% CI, 0.38 to 0.78; p=0.0007) and also found an association with progression-free survival (HR, 0.72; 95% CI, 0.54 to 0.96; p=0.026) in a homogenous group of 388 high-stage (stages III-IV using the International Federation of Gynecology and Obstetrics staging system), optimally debulked high-grade serous ovarian cancer patients. GTF2H5-silencing induced a decrease of the half maximal inhibitory concentration upon cisplatin treatment in GTF2H5-silenced ovarian cancer cells. CONCLUSION: Low levels of GTF2H5 are associated with enhanced prognosis in high-grade serous ovarian cancer patients and may contribute to cisplatin sensitization.
Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor/biosynthesis/genetics ; Cystadenocarcinoma, Serous/*genetics/metabolism/pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Kaplan-Meier Estimate ; Middle Aged ; Neoplasm Grading ; Neoplasm Proteins/biosynthesis/genetics ; Neoplasms, Glandular and Epithelial/*genetics/metabolism/pathology ; Ovarian Neoplasms/*genetics/metabolism/pathology ; Prognosis ; Transcription Factors/biosynthesis/*genetics ; Tumor Cells, Cultured

OBJECTIVEThe study intended to investigate the effect and mechanism of endoplasmic reticulum stress on cisplatin resistance in ovarian carcinoma.

METHODSRT-PCR and Western blot were used to test the expression of mTOR and Beclin1 mRNA and protein in ovarian cancer SKOV3 cells after saquinavir induction. MTT assay was used to analyze the influence of saquinavir on cisplatin sensitivity in SKOV3 cells.

RESULTSThe IC50 of SKOV3 cells was (5.490 ± 1.148) µg/ml. After induced by Saquinavair 10 µmol/L and 20 µmol/L, the IC50 of SKOV3 cells was increased to (11.199 ± 0.984) µg/ml and (14.906 ± 2.015) µg/ml, respectively. It suggested that the sensitivity of ovarian cancer cells to cisplatin was decreased significantly (P = 0.001). The expression of mTOR and Beclin1 mRNA and protein was significantly different among the five groups: the (Saquinavair+DDP) group of, Saquinavair group, LY294002 group, DDP group and control group (P < 0.001) . The expressions of mTOR and Beclin1 mRNA were highest in the (Saquinavair+DDP) group, 0.684 ± 0.072 and 0.647 ± 0.047, respectively; Secondly, the Saquinavair group, 0.577 ± 0.016 and 0.565 ± 0.037, respectively. The expressions of mTOR and Beclin1 proteins were also highest in the (Saquinavair+DDP) group, 0.624 ± 0.058 and 0.924 ± 0.033, respectively, followed by the Saquinavair group, 0.544 ± 0.019 and 0.712 ± 0.024. 3-MA inhibited the autophagy and restored cisplatin sensitivity in the SKOV3 cells after Saquinavir induced ER stress (P < 0.001).

CONCLUSIONSSaquinavir can effectively induce endoplasmic reticulum stress in SKOV3 cells. Endoplasmic reticulum stress can decrease the sensitivity to cisplatin in SKOV3 cells. The mechanism of the decrease of sensitivity to cisplatin in SKOV3 cells may be that ERS regulates cell autophagy through the mTOR and Beclin1 pathways. ERS of tumor cells and autophagy may become a new target to improve the therapeutic effect of chemotherapy and to reverse the drug resistance in tumor treatment.

Antineoplastic Agents ; pharmacology ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Autophagy ; drug effects ; Beclin-1 ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Drug Resistance, Neoplasm ; Endoplasmic Reticulum Stress ; drug effects ; Female ; HIV Protease Inhibitors ; pharmacology ; Humans ; Membrane Proteins ; genetics ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; RNA, Messenger ; Saquinavir ; pharmacology ; TOR Serine-Threonine Kinases ; genetics ; metabolism

Age of Onset ; Cystadenocarcinoma, Serous ; genetics ; metabolism ; pathology ; Female ; Genes, BRCA1 ; Genes, BRCA2 ; Humans ; Immunohistochemistry ; Neoplasm Staging ; Ovarian Neoplasms ; genetics ; metabolism ; pathology ; Receptors, Progesterone ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVETo investigate the effects of HE4 gene knockdown on the proliferation, adhesion and invasion of the ovarian cancer cells SKOV3.

METHODSThe knockdown of HE4 gene was performed by RNAi technology. The recombinant plasmids (pSUPER-HE4 shDNAs) were constructed and transfected into human ovarian cancer cells SKOV3. HE4 expression was then identified by real-time PCR and Western blot analysis. The invasion and adhesion ability of transduced cells were determined. In addition, cell proliferation and growth were analyzed by colonies formation assay.

RESULTSKnockdown of HE4 was achieved, and further confirmed by real-time PCR and Western blot. The proliferation of HE4-down-regulated cells was not affected, but the invasion ability of the transfected cells was reduced (P < 0.05) and the adhesion ability was also reduced to 27.3%.

CONCLUSIONHE4 expression is down-regulated effectively by the constructed HE4 shDNA, and thus knockdown of HE4 inhibits the adhesion and invasion of SKOV3 cells.

Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cystadenocarcinoma, Serous ; metabolism ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Humans ; Ovarian Neoplasms ; genetics ; metabolism ; pathology ; Plasmids ; Proteins ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; genetics ; metabolism ; Transfection

OBJECTIVETo investigate the correlation of ZNF217 gene expression with the biological behavior of human ovarian cancer HO-8910 cells.

METHODSThe expression of ZNF217 in ovarian carcinoma cell lines was detected by RT-PCR and Western blot, respectively. The biological behaviors of the transfectants were investigated by MTT, in vitro Boyden chamber and in vivo invasion assay, respectively.

RESULTSRT-PCR and Western blotting revealed that transfection of ZNF217 into the HO-8910 cells significantly increased their proliferation along with markedly enhanced in vitro and in vivo invasion and metastatic abilities. MTT assay showed that the proliferation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells and HO-8910 cells (P < 0.001). The Boyden chamber assay showed that the numbers of migrating pEGFP-N1-ZNF217/HO-8910, pEGFP-N1/HO-8910 and HO-8910 cells were (141.25 ± 13.91) cells/200×field, (82.50 ± 11.73) cells/200×field and (81.75 ± 12.12)cells/200×field, with a significant difference between them (F = 29.247, P < 0.001). The nude mouse experiment showed that the in vivo tumor formation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells (P < 0.001).

CONCLUSIONSZNF217 gene plays an important role in the invasion and metastasis of ovarian cancer. ZNF217 gene expression may be a useful marker indicating invasion and metastasis of ovarian cancer.

Animals ; Biomarkers, Tumor ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Female ; Humans ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Transplantation ; Ovarian Neoplasms ; metabolism ; pathology ; Plasmids ; RNA, Messenger ; metabolism ; Random Allocation ; Trans-Activators ; genetics ; metabolism ; Transfection ; Tumor Burden

Triptolide, a compound extracted from the traditional Chinese medicine preparation of Tripterygium wilfordii Hook F., has been reported to have anti-inflammatory and anti-cancer activities. However, its effect on ovarian cancer invasion is unknown. We observed that MMP7 and MMP19 expression increased in ovarian cancer tissue. Triptolide treatment inhibited the migration and invasion of ovarian cancer cells SKOV3 and A2780 at the concentration of 15 nM. We also observed that triptolide suppressed MMP7 and MMP19 promoter activity in a dose-dependent manner, down-regulating the expressions of these promoters on mRNA and protein level. Moreover, triptolide enhanced E-cadherin expression in ovarian cancer cells. In vivo, triptolide inhibited tumor formation and metastasis in nude mice, and suppressed MMP7 and MMP19 expression; it also enhanced E-cadherin expression in tumor in a dose-dependent manner. Over expression of MMP7 and MMP19, or suppression of E-cadherin expression partially abolished the inhibitory effect of triptolide on invasion of ovarian cancer cells. To summarize, triptolide significantly inhibited the migration and invasion of ovarian cancer cells by suppression of MMP7 and MMP19 and up-regulation of E-cadherin expression. This study shows that triptolide is a good candidate for the treatment of ovarian cancer and reduction of metastasis.
Animals ; Antineoplastic Agents, Alkylating/*pharmacology ; Cadherins/*genetics/metabolism ; Cell Line, Tumor ; Cell Movement/drug effects ; Cell Proliferation/drug effects ; Cystadenocarcinoma, Serous/*drug therapy/pathology ; Diterpenes/*pharmacology ; Epoxy Compounds/pharmacology ; Female ; Gene Expression Regulation, Enzymologic/drug effects ; Humans ; Matrix Metalloproteinase 7/genetics/*metabolism ; Matrix Metalloproteinases, Secreted/genetics/*metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Ovarian Neoplasms/*drug therapy ; Paclitaxel/pharmacology ; Phenanthrenes/*pharmacology ; Promoter Regions, Genetic ; Up-Regulation/drug effects ; Xenograft Model Antitumor Assays

Adaptor Proteins, Signal Transducing ; metabolism ; Adenocarcinoma, Clear Cell ; genetics ; metabolism ; pathology ; surgery ; Adenosine Triphosphatases ; metabolism ; Age Factors ; Carcinoma, Endometrioid ; genetics ; metabolism ; pathology ; surgery ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; metabolism ; pathology ; surgery ; Cystadenocarcinoma, Serous ; genetics ; metabolism ; pathology ; surgery ; DNA Mismatch Repair ; DNA Repair Enzymes ; metabolism ; DNA-Binding Proteins ; metabolism ; Endometrial Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Female ; Humans ; Mismatch Repair Endonuclease PMS2 ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; metabolism ; Neoplasms, Multiple Primary ; genetics ; metabolism ; pathology ; surgery ; Nuclear Proteins ; metabolism

Adenocarcinoma, Clear Cell ; etiology ; pathology ; Brenner Tumor ; etiology ; pathology ; Carcinosarcoma ; etiology ; pathology ; Cell Transformation, Neoplastic ; Cystadenocarcinoma, Serous ; etiology ; pathology ; Epithelial Cells ; metabolism ; pathology ; Female ; Genes, p53 ; Humans ; Mutation ; Neoplasms, Glandular and Epithelial ; etiology ; genetics ; metabolism ; pathology ; Ovarian Neoplasms ; etiology ; genetics ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; genetics ; metabolism

Animals ; Biomarkers, Tumor ; genetics ; metabolism ; Cell Nucleus Division ; Cystadenocarcinoma, Serous ; classification ; genetics ; metabolism ; pathology ; Female ; Humans ; Mutation ; Neoplasm Grading ; Ovarian Neoplasms ; classification ; genetics ; metabolism ; pathology ; Proto-Oncogene Proteins B-raf ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism

OBJECTIVEThe aim of this study was to investigate whether miR-449a, miR-449b and miR-192 family microRNAs play the same roles in p53 pathway as miR-34 family in ovarian cancer.

METHODSWild-type p53 ovarian carcinoma cell line A2780 cells were treated with genotoxic agent adriamycin. The reactivation of p53 was detected by Western blot. The expression of miR-449a/b, miR-34a, miR-34b, miR-34c, miR-192 and miR-194 were detected by real-time quantitative PCR. Mutant p53 ovarian cancer cell line SKOV3.ipl cells were transfected with pre-microRNAs and the cell-cycle changes were detected. The expression level of miR-449a/b, miR-34a, miR-34b, miR-34c, miR-192 and miR-194 in serous ovarian carcinomas of varying grade and stage were compared with real-time PCR.

RESULTSThe expressions of miR-449a/b, miR-34b and miR-34c were 19-fold to 21-fold elevated after p53 activation by genotoxic agent. Ectopic expression of miR-449b, as well as miR-34c, resulted in cell-cycle arrest in SKOV3.ipl cells. The expression of miR-449a/b was parallel with that of miR-34b, miR-34c, and were significantly lower in late stage and high-grade serous carcinomas than in the normal fallopian tube, early stage and low-grade serous carcinomas. The expression of miR-192, miR-194 and miR-34a did not show evident features in serous ovarian carcinomas and were much lower than miR-449a/b, miR-34b and miR-34c in normal fallopian tube.

CONCLUSIONSAs tumor-suppressor microRNAs, miR-449a/b, miR-34b and miR-34c cooperate and play important roles in p53 pathway. Their inactivation may contribute to the carcinogenesis and progression of serous ovarian carcinomas.

Adult ; Aged ; Antibiotics, Antineoplastic ; pharmacology ; Cell Cycle ; Cell Line, Tumor ; Cystadenocarcinoma, Serous ; genetics ; metabolism ; pathology ; Doxorubicin ; pharmacology ; Female ; Humans ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Neoplasm Grading ; Neoplasm Staging ; Ovarian Neoplasms ; genetics ; metabolism ; pathology ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; Transfection ; Tumor Suppressor Protein p53 ; metabolism