七氟醚对前列腺癌细胞比卡鲁胺抵抗的影响及其机制

Effect and mechanism of sevoflurane on the bicalutamide resistance of prostate cancer cells

目的研究七氟醚在体外对前列腺癌细胞比卡鲁胺抵抗的影响及其可能的相关机制。方法培养人前列腺癌细胞株LNCa P并通过比卡鲁胺诱导构建抵抗比卡鲁胺的LNCa P细胞株(LNCa P-RB),0%、1.7%、3.4%及5.1%气体浓度的七氟醚处理LNCa P(分别为对照组、低浓度组、中浓度组及高浓度组)及LNCa P-RB细胞(分别为对照抵抗组、低浓度抵抗组、中浓度抵抗组及高浓度抵抗组)6 h,CCK-8实验检测不同浓度七氟醚处理后LNCa P及LNCa P-RB细胞的比卡鲁胺半抑制浓度(IC50),实时荧光定量PCR技术(RT-q PCR)检测各组细胞中微小RNA-216a(miR-216a)的表达水平,免疫印迹实验(Western blot)检测各组细胞中E-cadherin及Snail的表达。结果与对照组相比,高浓度组比卡鲁胺IC50显著降低,中浓度组及高浓度组miR-216a及Snail表达显著降低,且E-cadherin表达显著升高;与对照抵抗组相比,中浓度抵抗组及高浓度抵抗组比卡鲁胺IC50均显著降低,且Snail表达显著降低,中浓度抵抗组及高浓度抵抗组E-cadherin表达显著升高,低浓度抵抗组、中浓度抵抗组及高浓度抵抗组miR-216a表达显著降低,差异均具有统计学意义(P<0.05)。结论七氟醚可下调前列腺癌细胞miR-216a的表达并抑制上皮间充质转化(EMT)进程,增强细胞比卡鲁胺的敏感性。

Objective To explore the effect and possible mechanism of sevoflurane on the bicalutamide resistance of prostate cancer cells. Methods The human prostate cancer cell line LNCa P was cultured and the LNCa P bicalutamide resistance（LNCa P-RB） cell line was constructed by bicalutamide induction. The gas including 0%,1.7%,3.4% and 5.1% sevoflurane were used to treat LNCa P（as the control group,low concentration group,medium concentration group and high concentration group） and LNCa P-RB（as the control resistance group, low concentration resistance group, medium concentration resistance group and high concentration resistance group） for 6 h. CCK-8 assay was used to detect the bicalutamide 50%inhibitory concentration（IC50） of each group. Real-time fluorescence quantitative PCR（RT-q PCR） was used to detect the expression of microRNA-216 a（miR-216 a）. Western blot was used to detect the expression of E-cadherin and Snail. Results Compared with the control group, the bicalutamide IC50 was significantly decreased in the high concentration group,and the expression of E-cadherin was significantly increased in the medium and high concentration groups, and the expression of Snail and miR-216 a was decreased in the medium and high concentration groups. Compared with the control resistance group,the bicalutamide IC50 and Snail expression were significantly decreased in the medium and high concentration resistance groups,and the expression of E-cadherin was significantly increased in the medium and high concentration resistance groups,and the expression of miR-216 a was significantly decreased in the medium and high concentration resistance groups. All these differences were statistically significant（P 〈 0. 05）.Conclusion Sevoflurane can down-regulate the expression of miR-216 a in prostate cancer cells and inhibit the process of epithelial mesenchymal transition（EMT）,and enhance cell sensitivity to bicalutamide.