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目的探讨RNA干扰卵巢癌细胞黏着班激酶(FAK)基因后,卵巢癌细胞对顺铂和紫杉醇的敏感性变化。方法构建特异性FAK-shRNA重组质粒转染HO-8910卵巢癌细胞;采用反转录PCR(RT-PCR)技术测定转染前后HO-8910细胞FAK mRNA表达情况;MTT比色法检测靶向沉默FAK后HO-8910细胞在顺铂和紫杉醇作用下的抑制情况,计算细胞半数抑制浓度(IC50值),比较细胞对顺铂和紫杉醇化疗敏感性变化。结果特异性FAK-shRNA-2434重组质粒转染卵巢癌细胞,荧光显微镜下观察其转染效率达70%~80%以上;转染48h后细胞FAK mRNA表达抑制率约为69.80%,与空白组和阴性组比较,差异均有统计学意义(P<0.05);顺铂和紫杉醇转染组细胞的IC50值分别为(0.017±0.007)μg/ml和(4.095±0.410)μg/ml,与空白组及阴性组比较,差异均有统计学意义(P<0.05)。结论 RNA干扰载体能有效抑制卵巢癌细胞FAK mRNA表达,并增强细胞对顺铂和紫杉醇化疗敏感性。
Objective To investigate the sensitivity of ovarian cancer cells to cisplatin and paclitaxel after RNAi interfered with the adhesion kinase (FAK) gene in ovarian cancer cells. Methods The specific FAK-shRNA recombinant plasmids were transfected into HO-8910 ovarian cancer cells. The expression of FAK mRNA in HO-8910 cells was detected by reverse transcription-polymerase chain reaction (RT-PCR) FAK HO-8910 cells in the cisplatin and paclitaxel inhibition, calculate the cell half inhibitory concentration (IC50 value), compare the cell sensitivity to cisplatin and paclitaxel changes. Results The transfection efficiency of FAK-shRNA-2434 recombinant plasmid was over 70% -80% after transfected into ovarian cancer cells. The inhibitory rate of FAK mRNA expression was 69.80% after transfected with FAK-shRNA-2434 for 48 hours, (0.017 ± 0.007) μg / ml and (4.095 ± 0.410) μg / ml, respectively, in the cells transfected with cisplatin and paclitaxel compared with the blank group (P <0.05) The differences between the two groups were statistically significant (P <0.05). Conclusion The RNA interference vector can effectively inhibit the expression of FAK mRNA in ovarian cancer cells and enhance the chemosensitivity of cells to cisplatin and paclitaxel.