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目的:分析miRNA-17~92基因簇过表达与卵巢癌SKOV3细胞紫杉醇耐药的关系,为卵巢癌化疗耐药的研究提供理论依据。方法:脂质体法将构建的p EGFP-N1-miR-17~92质粒转染入卵巢癌SKOV3细胞。Real time-PCR法及Western blot法分别检测转染前后miR-17~92及BIM、PTEN表达。结果:与SKOV3细胞相比,空质粒p EGFPN1转染的SKOV3-p EGFP-N1细胞miR-17~92含量0.970,与对照组比较差异无统计学意义(t=0.581,P=0.620);过表达质粒p EGFP-N1-miR-17~92转染的SKOV3-p EGFP-N1-miR-17~92细胞miR-17~92含量2.082,与对照组比较差异有统计学意义(t=-5.602,P=0.030);SKOV3-p EGFP-N1-miR-17~92细胞组的miR-17~92 mRNA相对表达量与SKOV3-p EGFP-N1细胞组比较,差异有统计学意义(t=-8.473,P=0.014)。不同浓度紫杉醇作用于转染p EGFP-N1-miR-17~92质粒后细胞,miR-17~92过表达使BIM蛋白表达下降,对PTEN蛋白表达的影响不明显。结论:miR-17~92基因簇与卵巢癌SKOV3细胞紫杉醇耐药有关。miR-~92基因簇过表达影响卵巢癌耐药的机制是通过下调BIM蛋白而非PTEN蛋白参与。
OBJECTIVE: To analyze the relationship between miRNA-17-92 gene cluster overexpression and paclitaxel resistance in ovarian cancer SKOV3 cells and to provide a theoretical basis for the study of chemoresistance in ovarian cancer. METHODS: The constructed p EGFP-N1-miR-17-92 plasmid was transfected into ovarian cancer SKOV3 cells by lipofectamine. Real time-PCR and Western blot were used to detect the expression of miR-17-92 and BIM, PTEN before and after transfection. Results: Compared with SKOV3 cells, the expression of miR-17-92 in SKOV3-p EGFP-N1 cells transfected with empty plasmid p EGFPN1 was 0.970, which showed no significant difference compared with control group (t = 0.581, P = 0.620) The miR-17-92 content of SKOV3-p EGFP-N1-miR-17-92 cells transfected with EGFP-N1-miR-17-92 plasmid was 2.082, which was significantly different from the control group (t = -5.602 , P = 0.030). The relative expression level of miR-17-92 mRNA in SKOV3-p EGFP-N1-miR-17-92 cell group was significantly different from that in SKOV3-p EGFP- N1 cell group (t = 8.473, P = 0.014). Different concentrations of paclitaxel on transfected pEGFP-N1-miR-17-92 plasmid, miR-17 ~ 92 overexpression decreased BIM protein expression, PTEN protein expression was not significantly affected. Conclusion: The miR-17 ~ 92 gene cluster is associated with paclitaxel resistance in ovarian cancer SKOV3 cells. The mechanism of overexpression of miR-92 gene affecting the drug resistance of ovarian cancer is through downregulation of BIM protein rather than PTEN protein.