RNA干扰抑制S100A8基因表达对Hec-1B细胞凋亡及体外药敏的影响  

The Effect on Apoptosis and Chemosensitivity of Hec-1B Endometrial Carcinoma Cells in vitro When S100A8 Gene Expression is Inhibited by RNA Interference

作　　者：黄带发[1] 林朝胜[1] 富伟能[2] 孙开来[2] 

机构地区：[1]沈阳军区总医院干二科,沈阳市110015 [2]中国医科大学遗传病教研室

出　　处：《中国肿瘤临床》2008年第8期441-444,共4页Chinese Journal of Clinical Oncology

基　　金：国家863子课题基金资助(编号:2002BA711A08-18)

摘　　要：目的:子宫内膜癌是女性生殖器最常见的恶性肿瘤之一,晚期及复发性患者预后差,对多种化疗药物耐药。本文探讨RNA干扰抑制S100A8基因表达对子宫内膜癌细胞Hec-1B凋亡及体外药敏的影响。方法:体外合成RNA干扰用S100A8siRNA,采用脂质体将S100A8siRNA转染到人子宫内膜癌细胞Hec-1B中,用RT-PCR、West-ern blot检测S100A8基因表达,TUNEL和流式细胞仪检测细胞凋亡变化,MTT法检测细胞抑制率。在转染S100A8siRNA后不同时间的Hec-1B细胞中分别加入2个浓度的顺铂,表柔比星,紫杉醇等药物,设立阴性对照,观察Hec-1B细胞体外生长的变化。结果:1)转染S100A8siRNA可抑制Hec-1B细胞的S100A8基因表达,该作用可维持10天,随着该基因表达被抑制,细胞凋亡增加,在第5天达最高峰,转染组细胞凋亡率(30.34±10.30)%,明显高于对照组(18.14±2.68)%,(P=0.01),而随着S100A8基因表达恢复,细胞凋亡率下降至转染前水平。2)Hec-1B细胞S100A8基因表达抑制后,顺铂,表柔比星,紫杉醇等药物抑制Hec-1B细胞生长作用明显增强。除40.0μmol/L表阿霉素(P=0.056)外,33.3μmol/L和66.5μmol/L顺铂,20.0μmol/L表阿霉素,5.0μmol/L和10.0μmol/L紫杉醇,与对照组比较,均显示对Hec-1B细胞生长抑制增强,存在显著差异(P<0.05)。结论:我们首次报道RNA干扰可抑制Hec-1B细胞S100A8基因表达,诱导该细胞凋亡,抑制S100A8基因表达使Hec-1B对化疗药物敏感性增加,提示该基因可能与子宫内膜癌耐药相关。To investigate the effect of inhibiting the S100A8 gene using siRNA on apoptosis and chemosensitivity of Hec-lB cells in vitro. Methods： Inhibited expression of S100A8 mRNA was achieved after transfection of a vector expressing S100A8 siRNA. Reverse transeriptase polymerase chain reaction （RT-PCR） and Western blot were used to detect the expression of S100A8 mRNA and protein. Apoptosis was detected by flow cytometry as well as fluoreseent staining. The inhibitory rates of Cisplatin, Epirubicin and Paclitaxel in Hec-1B cells were detected by MTT assay in vitro. Each drug was administered in two different concentrations. Untransfected Hec-1B cells were used as the control. Results： We found remarkable suppression of the expression of S100A8 in Hec-1B cells transfected with the S100A8 siRNA vector and the effect was maintained for 10 days. The rate of apoptosis was 30.34±10.30% in travlsfected cells and 18.14±2.68% in the controls （P=0.01）. The growth of transfected Hec-1B cells was inhibited remarkably by Cisplatin. Epirubiein and Paclitaxel. with a significant difference from that of the controls （P〈0.05）. Conclusion： RNA interferenee of the S100A8 gene can inhibit the expression of S100A8 in Hec-1B cells, inducing Hec-1B cell apoptosis and increasing the chemosensitivity of Hec-1B cells. S100A8 may be involved in the mechanism of multidrug resistance in Hec-1B cells.

关 键 词：S100A8基因 RNA干扰 肿瘤细胞 细胞调亡 药物敏感性 

分 类 号：R737.33[医药卫生—肿瘤]