CSN6对宫颈癌HeLa细胞与SiHa细胞自噬能力的影响研究  

The Effect of CSN6 on the Autophagy Ability of Cervical Cancer HeLa Cells and SiHa Cells

作　　者：李想[1] 于思琪 李彤[1] 薛朋旭 罗富海 LI Xiang;YU Si-qi;LI Tong;XUE Peng-xu;LUO Fu-hai(Faculty of Medicine,Jiamusi University,Jiamusi 154007,Heilongjiang,China)

机构地区：[1]佳木斯大学医学部,黑龙江佳木斯154007

出　　处：《医学信息》2020年第17期73-76,共4页Journal of Medical Information

基　　金：2020年黑龙江省大学生创新创业训练项目(编号:202010222066)。

摘　　要：目的探讨组成型光形态发生因子9信号复合体亚基6(CSN6)对宫颈癌HeLa细胞与SiHa细胞自噬能力的影响。方法购置宫颈癌HeLa细胞与SiHa细胞,并测定腺病毒中GFP与mRFP表达水平,标记、追踪LC3,示踪自噬流的水平;将购置的细胞放置在6孔板中进行培养,随机分为对照序列组、CSN6 siRNA组、空载体质粒组与CSN6过表达组,各组细胞均进行相应的处理(连续完成24 h干预);采用Western Blot法测定各组细胞中CSN6、LC3蛋白、Beclin1、Atg5表达水平,荧光显微镜下检测两种细胞中自噬体个数,采用四甲基偶氮唑蓝(MTT)法检测各组细胞干预后增殖抑制率。结果siCSN6组不同宫颈癌细胞中自噬体个数高于siCtrl组,差异具有统计学意义(P<0.05);siCSN6组CSN6、LC3蛋白、Beclin1、Atg5表达水平高于siCtrl组、siCSN6组及CSN6组,差异具有统计学意义(P<0.05);CSN6组CSN6、LC3蛋白、Beclin1、Atg5表达水平低于siCtrl组、siCSN6组,差异具有统计学意义(P<0.05);siCSN6组干预后2、6、12、24、48 h细胞增殖抑制率高于siCtrl组、siCSN6组及CSN6组,差异具有统计学意义(P<0.05)。结论转染CSN6能提高宫颈细胞的自噬水平,而过表达CSN6则能抑制自噬活性,均会对细胞增殖抑制率产生影响。Objective To investigate the effect of constitutive photomorphogenetic factor 9 signaling complex subunit 6(CSN6)on the autophagy ability of cervical cancer HeLa cells and SiHa cells.Methods Purchasing cervical cancer HeLa cells and SiHa cells,and determining the expression levels of GFP and mRFP in adenovirus,labeling and tracking LC3,and tracing the level of autophagic flux;the purchased cells were placed in a 6-well plate for culture and randomly divided into control sequence group,CSN6 siRNA group,empty vector plasmid group and CSN6 overexpression group.Cells in each group were treated accordingly(continuously completed 24 h intervention);Western Blot method was used to determine the CSN6,LC3 protein,the expression levels of Beclin1 and Atg5,the number of autophagosomes in the two types of cells were detected under a fluorescence microscope,and the proliferation inhibition rate of each group of cells after intervention was detected by the tetramethylazazole blue(MTT)method.Results The number of autophagosomes in different cervical cancer cells in the siCSN6 group was higher than that in the siCtrl group,the difference was statistically significant(P<0.05);the expression levels of CSN6,LC3 protein,Beclin1,Atg5 in the siCSN6 group were higher than those in the siCtrl group,siCSN6 group and CSN6 group,the difference was statistically significant(P<0.05);the expression levels of CSN6,LC3 protein,Beclin1,Atg5 in the CSN6 group were lower than those in the siCtrl and siCSN6 groups,the difference was statistically significant(P<0.05);The cell proliferation inhibition rate in the siCSN6 group was higher than that in the siCtrl group,siCSN6 group and CSN6 group at 2,6,12,24,and 48 h after intervention,the difference was statistically significant(P<0.05).Conclusion Transfection of CSN6 could increase the autophagy level of cervical cells,while overexpression of CSN6 could inhibit the autophagy activity,both of which would affect the cell proliferation inhibition rate.

关 键 词：组成型光形态发生因子9信号复合体亚基6 宫颈癌 自噬能力 增殖抑制率 

分 类 号：R737.33[医药卫生—肿瘤]