摘要
目的 探讨应用孕激素受体（PR）基因为靶标的siRNA抑制子宫内膜癌细胞Ishikawa该基因表达后,对瘤细胞化疗及内分泌治疗及化疗药物敏感性的影响.方法 设计以PR为靶标的siRNA1~3（均为100 μmol/L）转染入Ishikawa细胞,24 h后RT-PCR及Western blot法检测细胞PR蛋白及mRNA水平变化,评价siRNA的抑制效率.CCK8法检测未转染的单纯用药细胞和转染siRNA1～3后的细胞对紫杉醇、阿霉素、孕激素的药物敏感性,计算IC50值.应用AnnexinV/PI及Hochest33342染色各组细胞,流式细胞仪及荧光显微镜检测细胞凋亡率及细胞形态学变化.结果 转染以PR为靶标的siRNA3后,PR基因的mRNA及蛋白的表达下调75%以上.转染siRNA3的Ishikawa细胞紫杉醇、阿霉素、孕激素的IC50均较单纯用药细胞高,差异有统计学意义（P〉0.01）.结论 抑制子宫内膜癌细胞PR表达,可明显降低细胞对化疗药物及内分泌药物敏感性.
Objective To study the effects of progesterone receptor （PR） silencing on the chemo - drug sensiti- vity and endocrine therapy for endometrial cancer cell line Ishikawa using siRNA targeting PR. Methods The PR targeting siRNA was transfected into Ishikawa cells by lipofectamine. The mRNA and protein levels of PR were assessed by RT - PCR and WB 24 hours after transfection. The viability of Ishikawa cells treated with siRNA and paclitaxel, adriamycin or progestogen was assessed by CCK8 method to calculate the ICs0. After stained by Annexin V/PI and Hochest33342, the apoptosis ratio and cell morphological change were assessed with FCM and fluorescence microscope. Results The mRNA and protein of PR were reduced by more than 75% with PR targeting siRNA. Meanwhile, significant increase of IC50 and reduction of apoptosis ratio were observed in transfected groups than single drug groups （ P 〈 0. 05 ）. Conclusion PR targeting siRNA - induced PR silencing can reduce chemo - drug sensitivity and endocrine therapy for endometrial cancer cell line Ishikawa.
出处
《广东医学》
CAS
CSCD
北大核心
2013年第18期2753-2756,共4页
Guangdong Medical Journal
基金
广东省医学科研基金资助项目(编号:A2010333
B2010193)
