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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2021, Vol. 41 Issue (6): 4-12    DOI: 10.13523/j.cb.2101026
研究报告     
GTF2H2通过介导AKT信号通路影响肝癌细胞Hep3B的增殖和迁移*
欧阳琴,李艳萌,徐安健,周冬虎,李振坤,黄坚()
首都医科大学附属北京友谊医院科研实验中心 北京市临床医学研究所 北京 100875
GTF2H2 Affects the Proliferation and Migration of Hep3B Hepatocellular Carcinoma Cells by Mediating AKT Signal Pathway
OUYANG Qin,LI Yan-meng,XU An-jian,ZHOU Dong-hu,LI Zhen-kun,HUANG Jian()
Experimental Center, Beijing Friendship Hospital, Beijing Institute of Clinical Medicine, Beijing 100875, China
 全文: PDF(2099 KB)   HTML
摘要：

目的:探讨通用转录因子II H亚基2(GTF2H2)是否影响肝癌细胞Hep3B的增殖和迁移及其潜在的分子机制。方法:通过转染GTF2H2-siRNA构建GTF2H2敲低的Hep3B肝癌细胞模型;实时定量聚合酶链反应(q-RT-PCR)和蛋白质印迹实验检测肝癌细胞Hep3B的GTF2H2敲低效果;细胞计数实验(MTS)检测GTF2H2敲低的肝癌细胞Hep3B的增殖能力;Transwell细胞迁移实验检测GTF2H2敲低的肝癌细胞Hep3B的迁移能力;蛋白质印迹分析实验检测GTF2H2敲低后是否影响肿瘤相关分子信号通路。结果: GTF2H2敲低组的Hep3B细胞的增殖能力较对照组的Hep3B细胞增强,迁移能力亦有增强;蛋白质印迹实验显示GTF2H2敲低后,p-AKT通路蛋白的表达明显升高。结论:GTF2H2可能通过介导AKT分子信号通路,影响肝癌细胞Hep3B的增殖和迁移能力。
关键词： Hep3B;  GTF2H2;  p-AKT;  增殖;  迁移    
Abstract:

Objective: To investigate whether General transcription factor II subunit 2(GTF2H2) affects the proliferation and migration of Hep3B cells and the underlying molecular mechanism. Methods: The GTF2H2 knockdown Hep3B cell model was constructed by transfecting GTF2H2-siRNA. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the GTF2H2 knockdown effect in Hep3B cells. MTS cell proliferation assay kit was used to detect the proliferation ability of GTF2H2 knockdown Hep3B cells. The migration ability of GTF2H2 knockdown Hep3B cells was evaluated by cell Transwell assay. Western blotting was used to detect whether GTF2H2 knockdown affects tumor associated molecular signaling pathway. Results: The proliferation and migration ability of GTF2H2 knockdown Hep3B cells was stronger than that of the controls. Western blotting showed that the expression of p-AKT pathway protein in GTF2H2 knockdown Hep3B cells was significantly increased. Conclusion: GTF2H2 may affect the proliferation and migration ability of Hep3B cells by the regulation of the AKT molecular signal pathway.
Key words: Hep3B    GTF2H2    p-AKT    Proliferation    Migration
收稿日期: 2021-01-20 出版日期: 2021-07-06
ZTFLH:  Q819  
基金资助: * 国家自然科学基金(81650014)
通讯作者: 黄坚     E-mail: huangj1966@hotmail.com
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欧阳琴
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李振坤
黄坚

引用本文:

欧阳琴,李艳萌,徐安健,周冬虎,李振坤,黄坚. GTF2H2通过介导AKT信号通路影响肝癌细胞Hep3B的增殖和迁移*[J]. 中国生物工程杂志, 2021, 41(6): 4-12.

OUYANG Qin,LI Yan-meng,XU An-jian,ZHOU Dong-hu,LI Zhen-kun,HUANG Jian. GTF2H2 Affects the Proliferation and Migration of Hep3B Hepatocellular Carcinoma Cells by Mediating AKT Signal Pathway. China Biotechnology, 2021, 41(6): 4-12.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2101026    或    https://manu60.magtech.com.cn/biotech/CN/Y2021/V41/I6/4

上游引物 下游引物
GTF2H2-V1 TA-GTCCTCCTCCTGCTAGCT GGGTTTTGCATCCTGGTCAG
GTF2H2-V2 GCAGCCTTACAACTTGCGAT CGCGAACTTCTGCAGACAAT
GTF2H2-V3 ATTGTCTGCAGAAGTTCGCG AGCTAGCAGGAGGAGGACTA
GAPDH GAGTCAACGGATTTGGTCGT GACAAGCTTCCCGTTCTCAG
表1  GTF2H2-V1、GTF2H2-V2、GTF2H2-V3、GAPDH引物序列表
图1  构建GTF2H2敲低的肝癌细胞Hep3B细胞模型
GTF2H2-siRNAs GTF2H2-V1 GTF2H2-V2 GTF2H2-V3
t值 P值 t值 P值 t值 P值
GTF2H2-siRNA1 13.54 0.005 4 18.56 0.002 9 8.364 0.014 0
GTF2H2-siRNA2 20.76 0.002 3 23.47 0.001 8 5.026 0.037 4
GTF2H2-siRNA3 8.818 0.012 6 10.68 0.008 7 3.839 0.061 7
表2  GTF2H2-siRNAs处理Hep3B细胞后GTF2H2 mRNA表达量的统计值
图2  GTF2H2敲低后促进了Hep3B细胞的增殖能力
0H 24H 48H 72H 96H
t值 2.211 16.94 8.495 3.274 2.629
P值 0.069 <0.000 1 0.000 1 0.017 0.039 1
表3  不同时间对照组和GTF2H2敲低组Hep3B细胞增殖OD值统计表
图3  敲低GTF2H2后促进了肝癌细胞Hep3B的迁移能力
图4  敲低GTF2H2 后激活了p-AKT通路
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