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Objective: To observe the effect of CoCl on the cisplatin sensitivity of human ovarian cancer SKOV3 cells, and to clarify the possible mechanism. Methods: The SKOV3 cells were Cultured in vitro and randomly divided into control group, CoCT group, cisplatin (DDP) group and CoCT combined with DDP (combination) group. The cells in CoCL group were Cultured in normal cell medium for 20 h after cultured in 200 pmol • L CoCL for 4 h, the cells in DDP group were cultured in normal cell medium containing 10 mg • L DDP for 24 h, and the cells in combination group were cultured in 10 mg • L DDP for 20 h after cultured in 200 //mol • L CoCl • for 4 h. The survival rates of SKOV3 cells in various groups were detected by MTT method, and the positive expression intensities of hypoxia-inducible factor-1 a ( HIF-la) and inducible nitric oxide synthase (iNOS) in the cells in various groups were detected by immunofluorescence method. Rhod 2-AM fluorescence probe was used to observe the levels of Ca in mitochondria in the cells in various groups. Western blotting method was used to observe the expression levels of cytochrome C (cyto C) cysteinyl aspartasc 3 (caspasc 3) and cleaved cysteinyl aspartasc 3 (cleaved caspase 3). Muse apoptosis assay kit was used to detect the apoptotic rates of cells in various groups. Results: Compared with control group, the survival rate of the cells in CoCI group had no significant change (P> 0.05). and the survival rates of the cells in DDP and combination groups were decreased ( P0. 05) . and the expression levels of cyto C. caspase 3 and cleaved caspase 3 in DDP group were increased significantly ( P < 0.05); comparexl with DDP group, they were lower than those in combination group ( P<0. 05). Comparexl with control group∗ the apoptotic rate of SKOV3 cells in DDP group was increased significantly (P<.0. 05); the apoptotic rate of SKOV3 cells in combination group was lowe'r than that in DDP group (P<0. 05). Conclusion: CoCI can redece the mitochondrial apoptosis of human ovarian cancer SKOV3 cells by inhibiting the DDP-inducexl enhancement of iNOS expression and dccrease the sensitivity of SKOV3 cells to cisplatin.
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Objective The objective of this study was to investigate the effect of microRNA-409-3p(miRNA-409-3p) expression on proliferation of cervical cancer Hela cells and chemotherapy sensitivity of cisplatin. Methods HeLa cells were divided into normal control,miRNA-409-3p mimic and RNA control groups. The mRNA expression of miRNA-409-3p was detected by RT-qPCR. The cell viability was detected by CCK-8 assay,and the expression of Fip200,LC3 and Caspase-3 was detected by Western blot. Results The relative expression level of microRNA-409-3p mRNA in cervical cancer tissues was significantly lower than that in normal cervical tissues and adjacent tissues(P<0. 05). After transfection of microRNA-409-3p mimic,the relative ex-pression level of microRNA-409-3p mRNA in the microNA-409-3p mimic group was significantly up-regulated compared with the normal control group(P<0. 05). The relative expression level of miRNA-409-3p mRNA in the RNA control group was not dif-ferent from that in the normal control group(P>0. 05). The proliferative rate of HeLa cells int the microRNA-409 mimic group was significantly lower than that in normal control and RNA control groups(P<0. 05). After cisplatin treatment,the proliferation of Hela cells was significantly inhibited in the miRNA-409-3p mimic group(P<0. 05);the expression of Fip200 protein and LC3II/LC3I ratio in the microRNA-409-3p mimic group were significantly lower than those in the normal control and RNA control groups(P<0. 05). Conclusion The low expression of microRNA-409-3p in cervical cancer tissue may be related to the occurrence and de-velopment of cervical cancer. Upregulation of microRNA-409-3p level can inhibit the proliferation of HeLa cells and increase the sensitivity of HeLa cells to cisplatin.
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Objective @# To investigate the effect of mitochondrial fission protein 1 (FIS1) on apoptosis and cisplatin resistance in tongue squamous cell carcinoma (TSCC) cells.@*Methods @#The squamous cell carcinoma cell lines SCC9 and CAL27 were used to detect the mRNA and protein levels of FIS1 after cisplatin treatment, the knockdown and overexpression of FIS1 of SCC9 and CAL27 with or without cisplatin treatment were accomplished through small interfering RNA (siRNA) and plasmid, respectively. The mitochondrial division state in cells was detected by mitochondrial staining, and the apoptosis state of cells was detected by TUNEL, flow cytometry and Caspase 3/7.@*Results@#FIS1 protein expression in tongue squamous carcinoma cells treated with cisplatin was increased, but the mRNA level did not change. Silencing of FIS1 expression reduced mitochondrial division and apoptosis in squamous cell carcinoma cells treated with cisplatin, whereas overexpression of FIS1 exhibited the opposite effects. The percentage of dividing mitochondria, the number of apoptotic cells and the activity of Caspase 3/7 in SCC9 and CAL27 cells were significantly different before and after modulation of FIS1 expression (P < 0.05). @*Conclusion@#FIS1 is involved in the regulation of cisplatin chemotherapy sensitivity in tongue squamous cell carcinoma and can be used as a new target for improving the sensitivity of cisplatin chemotherapy in oral squamous cell carcinoma.
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Objective:To observe the effect of CoCl2on the cisplatin sensitivity of human ovarian cancer SKOV3cells, and to clarify the possible mechanism.Methods:The SKOV3cells were cultured in vitro and randomly divided into control group, CoCl2 group, cisplatin (DDP) group and CoCl2 combined with DDP (combination) group.The cells in CoCl2group were cultured in normal cell medium for 20hafter cultured in 200μmol·L-1 CoCl2for 4h, the cells in DDP group were cultured in normal cell medium containing 10mg·L-1 DDP for 24h, and the cells in combination group were cultured in 10mg·L-1 DDP for 20hafter cultured in 200μmol·L-1 CoCl2for 4h.The survival rates of SKOV3cells in various groups were detected by MTT method, and the positive expression intensities of hypoxia-inducible factor-1α (HIF-1α) and inducible nitric oxide synthase (iNOS) in the cells in various groups were detected by immunofluorescence method.Rhod 2-AM fluorescence probe was used to observe the levels of Ca2+in mitochondria in the cells in various groups.Western blotting method was used to observe the expression levels of cytochrome C (cyto C) , cysteinyl aspartase 3 (caspase 3) and cleaved cysteinyl aspartase 3 (cleaved caspase 3) .Muse○R apoptosis assay kit was used to detect the apoptotic rates of cells in various groups.Results:Compared with control group, the survival rate of the cells in CoCl2group had no significant change (P＞0.05) , and the survival rates of the cells in DDP and combination groups were decreased (P＜0.05) ;the survival rate in combination group was higher than that in DDP group (P＜0.05) .Compared with control group, the positive expression intensities of HIF-1αin CoCl2and combination groups were increased (P＜0.05) .Compared with control group, the positive expressions of iNOS in DDP and combination groups were increased (P＜0.05) .The Ca2+levels in the cells in DDP group and combination groups were higher than that in control group (P＜0.05) and the Ca2+level in DDP group was higher than that in combination group (P＜0.05) .Compared with control group, the expression levels of cyto C, caspase 3and cleaved caspase 3proteins in the SKOV3cells in CoCl2group had no significant changes (P＞0.05) , and the expression levels of cyto C, caspase 3and cleaved caspase 3in DDP group were increased significantly (P＜0.05) ;compared with DDP group, they were lower than those in combination group (P＜0.05) .Compared with control group, the apoptotic rate of SKOV3cells in DDP group was increased significantly (P＜0.05) ;the apoptotic rate of SKOV3cells in combination group was lower than that in DDP group (P＜0.05) .Conclusion:CoCl2can redece the mitochondrial apoptosis of human ovarian cancer SKOV3cells by inhibiting the DDP-induced enhancement of iNOS expression and decrease the sensitivity of SKOV3cells to cisplatin.
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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.