LRP16影响宫颈癌Siha细胞对化疗药物的敏感性

Significance of Chemotherapy Sensitivity of the Expression Level of the Human LRP16 Gene in Siha Cells

目的:探讨抑制LRP16的表达对宫颈癌Siha细胞的化疗药物敏感性的影响。方法:将抑制LRP16表达的小干扰RNA:negativecontrol-si RNA(NC)、si RNA-374(si374)转染入Siha宫颈鳞癌细胞系中,通过顺铂(DDP)和紫杉醇(TAX)的处理后,采用CCK-8检测不同浓度紫杉醇、顺铂作用宫颈癌细胞系Siha48 h后,计算出细胞被抑制一半时顺铂、紫杉醇的药物浓度(IC50);使用Hoechst33342染色观察细胞凋亡,采用流式细胞仪检测顺铂IC50作用Siha细胞48小时后的细胞凋亡情况,紫杉醇IC50作用Siha细胞之后的细胞周期分布情况。结果:CCK-8检测转染的Siha细胞增殖活性受到抑制,Hoechst33342染色观察转染的Siha细胞凋亡明显增加,流式细胞仪检测凋亡显示,si374+顺铂的早期凋亡率22.15±2.24,NC+顺铂12.45±2.72,流式细胞仪检测周期显示G2/M(%),si374+紫杉醇29.94±1.87,NC+紫杉醇17.66±2.32。结论:LRP16基因表达下调之后,抑制Siha细胞的增殖、促进其凋亡,使细胞周期滞留于G2/M期,从而提高Siha细胞的化疗敏感性。

Objective： To investigate the effect of the inhibition of expression of LRP16 in Siha cervical cancer cells on chemotherapeutic drug sensitivity. Methods： Cervical squamous carcinoma cell （Siha cell） was transfected with small interfering RNA which was as negative control[siRNA（NC）] and small interfering RNA which was inhibited the expression of LRP16[siRNA-374（si374）], then was treated with cisplatin and paclitaxel. Siha cell was detected by CCK-8 with the treatment of different concentrations of cisplatin and paclitaxel after 48 hours. The drug （cisplatin or paclitaxel） concentration was calculated when Siha cells were half inhibited; Cell morphology was observated by Hoechst33342 staining. Flow cytometry was utilized to detect the apoptosis of Siha cells after the treatment of cisplatin ICS0 48 hours. G2/M distributionof the cell cycle was detected by flow cytometry after the treatment of paclitaxel IC50 48 hours. Results： The proliferative activity of transfected Siha was inhibited by CCK-8 detection and the apoptosis of transfected Siha was increased significantly when detectedby Hoechst33342.The early apoptosis rate of siLRP 16＋cisplatin was 22.15± 2.24, while the rate ofNC＋cisplatin was 12.45±2.72 by flow cYtometry. The G2/M distribution rate ofsiLRP16＋paclitaxel was 29.94±1.87, while NC＋ paclitaxel was17.66± 2.32. Conclusions： After the down-regulated expression ofLRP16 gene,the proliferation of Siha cells was inhibited, the apoptosis of Siha was induced and the cell cycle remained in the G2/M phase, thereby enhancing chemosensitivity of Siha cells.