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LRP16影响宫颈癌Siha细胞对化疗药物的敏感性
引用本文:于丹,伍志强,梅倩,白桦,韩为东,孟元光.LRP16影响宫颈癌Siha细胞对化疗药物的敏感性[J].现代生物医学进展,2015,15(20):3850-3854.
作者姓名:于丹  伍志强  梅倩  白桦  韩为东  孟元光
作者单位:解放军总医院
基金项目:国家自然科学基金项目(81071617,81001184)
摘    要: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。

关 键 词:LRP16  Siha  细胞增殖  细胞凋亡  细胞周期

Significance of Chemotherapy Sensitivity of the Expression Level of the Human LRP16 Gene in Siha Cells
Abstract: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 controlsiRNA(NC)] and small interfering RNA which was inhibited the expression of LRP16siRNA-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 IC50 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 siLRP16+cisplatin was 22.15± 2.24, while the rate of NC+ cisplatin was 12.45± 2.72 by flow cytometry. The G2/Mdistribution rate of siLRP16+paclitaxel was 29.94± 1.87, while NC+ paclitaxel was17.66± 2.32.Conclusion:After the down-regulated expression of LRP16 gene,the proliferation of Siha cells was inhibited, the apoptosis of Siha was induced and the cell cycle remained in the G2/Mphase, thereby enhancing chemosensitivity of Siha cells.
Keywords:LRP16  Siha  Cell proliferation  Cell apoptosis  Cell cycle
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