内质网相关降解蛋白Derlin-1对宫颈癌HeLa细胞生长的影响  

Effect of endoplasmic reticulum associated degradation protein Derlin-1 on proliferation of cervical cancer cell line He La in vitro

作　　者：吕娟[1] 李婧[1] 张春梅[1] 郭冬丽[1] 王赞宏[2] 

机构地区：[1]山西医科大学附属山西大医院妇产科,太原030001 [2]山西医学科学院山西大医院妇产科

出　　处：《山西医科大学学报》2015年第10期977-981,共5页Journal of Shanxi Medical University

基　　金：山西省科技产业化环境建设基金资助项目(2014071011)

摘　　要：目的采用RNAi技术抑制Derlin-1基因的表达,探讨其表达降低后对宫颈癌He La细胞生长的影响。方法合成针对Derlin-1基因的干扰片段,构建重组质粒,用脂质体法将重组质粒转染人宫颈癌He La细胞,并筛选出稳定表达株,作为siRNA-Derlin-1组,选取空质粒转染后的He La细胞为空质粒组,未做任何处理的He La细胞为空白对照组,采用RT-PCR检测重组质粒对He La细胞Derlin-1 mRNA的影响,Western blot检测其对Derlin-1蛋白表达的影响,MTT法分析其对细胞增殖的影响,ELISA法检测其对细胞凋亡的影响,同时检测凋亡因子caspase-3的表达。结果与空质粒组和空白对照组相比,siRNA-Derlin-1组He La细胞中Derlin-1 mRNA和蛋白的表达显著降低(P<0.05),MTT法显示siRNA-Derlin-1组宫颈癌He La细胞增殖能力显著低于空质粒组和空白对照组(P<0.05),同时siRNA-Derlin-1组凋亡细胞增加,凋亡核心蛋白caspase-3表达增加。结论抑制内质网相关降解蛋白Derlin-1的表达能有效降低宫颈癌He La细胞的增殖,促进凋亡。Objective To investigate the effect of Derlin-1 gene inhibition mediated by RNA interference on the proliferation of cervical cancer He La cells in vitro. Methods Recombinant plasmids for RNA interference of Derlin-1 gene were constructed and transfected into He La cells via lipofectamine 2000,which were named as siRNA-Derlin-1 group. Meanwhile,He La cells transfected with empty plasmids were chosen as empty plasmid group,and the untreated He La cells were selected as blank control group. The mRNA and protein expression of Derlin-1 in the stably transfected cells was detected by RT-PCR and Western blot. The changes of cell proliferation were detected by MTT assay. Enzyme-linked immunosorbent assay（ ELISA） was employed to observe the effect of transfection on the apoptosis. The expression of caspase-3 in the transfected cells was also detected by RT-PCR and Western blot. Results Compared with empty plasmids group and blank control group,the mRNA and protein expression levels of the He La cells in Derlin-1-siRNA group were significantly decreased（ P 〈0. 05）,and the cell proliferation was inhibited（ P〈 0. 05）. The apoptosis cells were increased in Derlin-1-siRNA group and the expression of caspase-3 was effectively promoted compared with empty plasmids group and blank control group（ P〈 0. 05）. Conclusion Derlin-1 gene could be highly expressed in cervical cancer and RNA interference-mediated Derlin-1silencing can strongly inhibit the proliferation and induce apoptosis in He La cells.

关 键 词：Derlin-1 宫颈癌 HELA RNA干扰 

分 类 号：R737.33[医药卫生—肿瘤]