内质网相关降解蛋白Derlin-1对宫颈癌C33A细胞自噬通路的影响  

Effect of endoplasmic reticulum-related degradation protein Derlin-1 on autophagy pathway of cervical cancer C33A cells

作　　者：任松洁[1] 吕娟 李婧[1] 王赞宏[1] REN Songjie;Lü Juan;LI Jing;WANG Zanhong(Department of Obstetrics and Gynecology,Affiliated Shanxi Bethune Hospital of Shanxi Medical University,Taiyuan 030032,China;Department of Obstetrics and Gynecology,General Hospital of Shanxi Jincheng Anthracite Mining Group Co.,Ltd)

机构地区：[1]山西医科大学附属白求恩医院妇产科,太原030032 [2]山西晋城无烟煤矿业集团有限责任公司总医院妇产科

出　　处：《山西医科大学学报》2020年第3期198-202,共5页Journal of Shanxi Medical University

基　　金：山西省应用基础研究项目(201601D102060);山西省科技成果转化引导专项项目(201604D132043)。

摘　　要：目的探讨内质网相关降解蛋白Derlin 1对宫颈癌C33A细胞自噬通路的影响。方法采用脂质体将HPV16 E6真核表达质粒pcDNA3.1(-)-HPV16 E6、空质粒pcDNA3.1(-)转染人宫颈癌C33A细胞,并筛选出稳定表达株;采用Western blot检测HPV16 E6过表达对HPV16 E6蛋白、内质网应激标志物GRP78蛋白、Derlin-1蛋白,自噬蛋白Beclin 1、P62表达的影响。采用脂质体将Derlin-1的干扰质粒PSUPER-siDerlin-1和空质粒PSUPER转染稳定表达HPV16 E6的C33A细胞;Western blot检测Derlin-1干扰质粒对Derlin-1蛋白、自噬蛋白Beclin 1、P62表达的影响。结果HPV16 E6真核表达质粒pcDNA3.1(-)-HPV16转染成功。与转染空质粒pcDNA3.1(-)和未转染的细胞比较,转染pcDNA3.1(-)-HPV16 E6后HPV16 E6蛋白过表达可以导致内质网应激,使GRP78蛋白、Derlin-1蛋白表达增加,同时使自噬蛋白Beclin 1表达增加,P62蛋白表达降低,差异均有统计学意义(P<0.05)。Derlin-1干扰质粒PSUPER-siDerlin-1转染成功。与转染空质粒PSUPER和未转染的细胞比较,PSUPER-siDerlin-1转染后Derlin-1蛋白表达降低,自噬蛋白Beclin 1、P62蛋白表达增加,差异均有统计学意义(P<0.05)。结论HPV16 E6蛋白过表达可以导致内质网应激,抑制Derlin-1蛋白表达可以阻断自噬溶酶体通路,Derlin-1可能是泛素—蛋白酶体通路和自噬—溶酶体通路之间的对话分子。Objective To investigate the effect of ER-related degradation protein Derlin 1 on autophagy pathway of cervical cancer C33A cells.Methods The eukaryotic expression plasmid pcDNA3.1(-)-HPV16 E6 and empty plasmid pcDNA3.1(-)were transfected via lipofectamine into human cervical cancer C33A cells,respectively,and the stable expression strains were screened out.And then Western blot was employed to detect the expression of HPV16 E6 protein,endoplasmic reticulum stress marker GRP78 protein,Derlin-1 protein,autophagic proteins Beclin 1 and P62.The interference plasmid PSUPER-siDerlin-1 and empty plasmid PSUPER were transfected via lipofectamine into C33A cells stably expressing HPV16 E6,and then Western blot was used to detect the expression of Derlin-1,Beclin-1 and P62.Results HPV16 E6 eukaryotic expression plasmid pcDNA3.1(-)-HPV16 was successfully transfected.Compared with the cells transfected with pcDNA3.1(-)and non-transfected cells,ER stress was induced in cells transfected with pcDNA3.1(-)-HPV16 E6,the expression of GRP78 and Derlin-1 proteins,and the expression of Beclin protein were increased(P<0.05),while the expression of P62 protein was decreased(P<0.05).Compared with the cells transfected with empty plasmid and untransfected cells,the expression of Derlin-1 protein was decreased in cells transfected with PSUPER-siDerlin-1(P<0.05),while the expression of Beclin-1 and P62 proteins was increased(P<0.05).Conclusion Overexpression of HPV16 E6 protein can lead to endoplasmic reticulum stress,and inhibition of Derlin-1 protein expression can block the autophagy-lysosomal pathway,so Derlin-1 may be a crosstalk molecule between ubiquitin-proteasome pathway and autophagy-lysosomal pathway.

关 键 词：内质网应激 Derlin-1 自噬 人乳头瘤病毒 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤]