肝癌Hep3B细胞蛋白磷酸酶2A的癌性抑制因子表达水平对凋亡素诱导凋亡的作用

doi:10.13481/j.1671-587x.20140210

吉林大学学报(医学版)

肝癌Hep3B细胞蛋白磷酸酶2A的癌性抑制因子表达水平对凋亡素诱导凋亡的作用

彭传梅¹，崔映波²，童淑芬²，梁蕾蕾²，李小洁²，狄勇²

（1.云南省昆明市延安医院检验科，云南昆明650051；2.昆明医科大学基础医学院生物化学与分子生物学系，云南昆明650500）

收稿日期:2013-07-05 出版日期:2014-03-28 发布日期:2014-03-28
通讯作者: 狄勇 E-mail:yongdi516@hotmail.com
作者简介:彭传梅（1980-），女，湖北省宜昌市人，主治医师，医学硕士，主要从事骨髓细胞学研究。
基金资助:
云南省科技厅-昆明医学院联合专项基金资助课题（2011FB241）

Effect of expression level of cancerous inhibitor of protein phosphatase 2A in hepatoma HepB3 cells on apoptosis induced by Apoptin

PENG Chuan-mei¹,CUI Ying-bo²,TONG Shu-fen²,LIANG Lei-lei²,LI Xiao-jie²,DI Yong²

(1.Clinical Laboratory,Yan’an Hospital of Kunming City,Yunnan Province,Kunming 650051,China;2.Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Kunming Medical Universiry,Kunming 650500,China)

Received:2013-07-05 Online:2014-03-28 Published:2014-03-28

摘要/Abstract

摘要：

目的：探讨肝癌HepB3细胞中蛋白磷酸酶2A的癌性抑制因子（CIP2A）的表达水平对凋亡诱导凋亡效率的影响，为研究凋亡素特异性诱导肿瘤细胞凋亡的机制提供依据。方法：定制合成CIP2A特异性siRNA，构建真核表达质粒pcDNA3-HA-Apoptin。以pcDNA3-HA-Apoptin和CIP2A siRNA共转染HepB3细胞作为实验组，转染pcDNA3-HA-Apoptin和CIP2A scrambled siRNA的HepB3细胞作为阳性对照组，转染pcDNA3和CIP2A siRNA的HepB3细胞作为阴性对照组，转染pcDNA3和CIP2A scrambled siRNA的HepB3细胞作为空白对照组。Western blotting法检测各组HepB3细胞中CIP2A和HA-Apoptin表达水平；流式细胞术检测各组HepB3细胞凋亡率；免疫荧光法检测各组HA-Apoptin在HepB3细胞核、质分布。结果：Western blotting法检测，与阴性对照组比较，实验组HepB3细胞内中CIP2A表达水平显著下降（P<0.001）；Western blotting和流式细胞术检测，与阳性对照组比较,实验组HepB3细胞凋亡率显著降低（P<0.01）；免疫荧光法检测，与阳性对照组比较，实验组HA-Apoptin在HepB3细胞质内的分布增多。结论：肝癌HepB3细胞中CIP2A表达水平下调降低了凋亡素诱导的凋亡。

关键词: 肝肿瘤, 蛋白磷酸酶2A的癌性抑制因子, 凋亡素, 细胞凋亡

Abstract:

bstract:Objective
To explore the influence of the expression level of cancerous inhibitor of protein phosphatase 2A(CIP2A) in the hepatoma HepB3 cell in the apoptosis induced by Apoptin,and to provide basis for studying the mechanism of Apoptin in the apoptin-specific induction of tumor cells.Methods CIP2A specific siRNA was synthesized and the eukaryotic expression plasmid pcDNA3-HA-Apoptin was constructed.CIP2A siRNA and pcDNA3-HA-Apoptin were co-transfected into HepB3 cells as experiment group,the HepB3 cells co-transfected with pcDNA3-HA-Apoptin and CIP2A scrambled siRNA were regarded as positive control group,the HepB3 cells co-transfected with pcDNA3 and CIP2A siRNA were regarded as negative control group,and the HepB3 cells co-transfected with pcDNA3 and CIP2A scrambled siRNA were regarded as blank control group.Western blotting method was used to detect the expression levels of CIP2A and HA-Apoptin in HepB3 cells;the apoptotic rates of HepB3 cells in different groups were detected by flow cytometry;immunofluorescence assay was utilized to determine the distribution of Apoptin in nucleus or cytoplasm in HepB3 cells.Results The Western blotting results showed that compared with negative control group,the expression level of CIP2A in the HepB3 cells in experiment group was decreased（P<0.001）.The Western blotting and flow cytometry assay results showed that the apoptotic rate of HepB3 cells in experiment group was significantly reduced compared with positive control group（P<0.05）.The immunofluorescence assay results indicated that the HA-Apoptin distribution in cytoplasm of the HepB3 cells in experiment group was increased compared with positive control group.Conclusion The downregulating of the expression level of CIP2A in HepB3 cells decreases the induction effect of Apoption on apoptosis.

Key words: liver neoplasms, cancerous inhibitor of protein phosphatase 2A, Apoptin, apoptosis

中图分类号:

R735.7

彭传梅，崔映波，童淑芬，梁蕾蕾，李小洁，狄勇. 肝癌Hep3B细胞蛋白磷酸酶2A的癌性抑制因子表达水平对凋亡素诱导凋亡的作用[J]. 吉林大学学报(医学版), 2014, 40(02): 261-265.

PENG Chuan-mei,CUI Ying-bo,TONG Shu-fen,LIANG Lei-lei,LI Xiao-jie,DI Yong. Effect of expression level of cancerous inhibitor of protein phosphatase 2A in hepatoma HepB3 cells on apoptosis induced by Apoptin[J]. Journal of Jilin University Medicine Edition, 2014, 40(02): 261-265.

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肝癌Hep3B细胞蛋白磷酸酶2A的癌性抑制因子表达水平对凋亡素诱导凋亡的作用

Effect of expression level of cancerous inhibitor of protein phosphatase 2A in hepatoma HepB3 cells on apoptosis induced by Apoptin

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