靶向干扰TAGLN表达对HBV阳性肝癌细胞生物学行为的影响及机制初探 <sup>*</sup>

doi:10.13523/j.cb.20191102

中国生物工程杂志

2019, Vol. 39

Issue (11): 13-21 DOI: 10.13523/j.cb.20191102

研究报告

靶向干扰TAGLN表达对HBV阳性肝癌细胞生物学行为的影响及机制初探 ^*

徐燕¹,刘正芸^1,²,张琬棂¹,王盛羽^1,²,王欢^1,^2,^**(

)

1 遵义医科大学医学与生物学研究中心遵义 563099
2 贵州省普通高等学校传染病与生物安全特色重点实验室遵义 563099

Effect of Targeted Interference with TAGLN Expression on Biological Behavior of HBV-Positive Hepatocellular Carcinoma Cells and Its Mechanisms

XU Yan¹,LIU Zheng-yun^1,²,ZHANG Wan-ling¹,WANG Sheng-yu^1,²,WANG Huan^1,^2,^**(

)

1 Research Center for Medical and Biology,Zunyi Medical University, Zunyi 563099, China
2 Key Laboratory of Infectious Disease & Biosafety, Provincial Department of Education, Zunyi 563099, China;

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摘要：

目的探究TAGLN对HBV阳性肝癌细胞HepG2.2.15生物学行为的影响及可能的作用机制。方法免疫组化法和Western blot检测TAGLN在HBV阳性和HBV阴性肝癌组织及细胞中的表达差异;用TAGLN干扰慢病毒感染HepG2.2.15细胞,通过嘌呤霉素筛选干扰TAGLN表达的稳定表达细胞系,Western blot验证干扰效率; CCK-8法和克隆形成实验检测干扰TAGLN表达对HepG2.2.15细胞增殖能力的影响;Transwell实验检测干扰TAGLN表达对HepG2.2.15细胞迁移和侵袭的影响;Western blot检测PI3K、p-PI3K、AKT以及p-AKT的表达。结果 TAGLN在HBV阳性肝癌组织及细胞中的表达高于HBV阴性肝癌组织和细胞(P<0.01);干扰TAGLN表达能抑制HepG2.2.15细胞增殖、克隆形成能力、迁移和侵袭(P<0.01);降低HepG2.2.15细胞中PI3K和AKT(P<0.01)及p-PI3K和p-AKT(P<0.05)的表达。结论在肝癌组织中,HBV感染能增加TAGLN的表达;干扰TAGLN表达后HepG2.2.15细胞的增殖能力、克隆形成能力、迁移和侵袭的能力减弱,其机制可能与PI3K及AKT的表达减少有关。

关键词： 转凝蛋白; 肝细胞癌; 乙肝病毒; AKT; PI3K

Abstract:

Objective: To invest igate the effect of TAGLN on the biological behavior of HBV-positive hepatocellular carcinoma cell line HepG2.2.15 and its underlying mechanism.Methods: Immunohistochemistry and Western blot were used to detect the expression levels of TAGLN in HBV-positive and HBV-negative hepatocellular carcinoma tissues and cells; HepG2.2.15 cells were infected with TAGLN interfering lentiviruses, and stable expression cell lines were screened by purinomycin. Then transfection efficiency was confirmed by Western blot; CCK-8 and clone formation assay were used to detect the proliferation of HepG2.2.15 cells. While cell migration and invasion capabilities were determined by Transwell assay. The expression levels of PI3K, p-PI3K, AKT and p-AKT were examined by Western blot.Results: The expression of TAGLN in HBV-positive hepatocellular carcinoma tissues and cells was higher than that in HBV-negative hepatocellular carcinoma tissues and cells(P<0.01); Knockdown of TAGLN could effectively inhibit cell proliferation, migration and invasion (P<0.01), as well as down-regulate the expression of PI3K, AKT(P<0.01), p-PI3K and p-AKT (P<0.05).Conclusion: HBV infection can increase the expression of TAGLN in hepatocellular carcinoma. After interfering with TAGLN expression, the proliferation, cloning, migration and invasion of HepG2.2.15 cells decreased, which may be related to the decrease of PI3K and AKT expression.

Key words: Transgelin HCC HBV AKT PI3K

收稿日期: 2019-04-14 出版日期: 2019-12-17

ZTFLH:

R373

基金资助: * 国家自然科学基金(81360261);* 贵州省教育厅(黔教合KY字[2015]331)

通讯作者: 王欢 E-mail: wanghuan928@163.com

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引用本文:

徐燕,刘正芸,张琬棂,王盛羽,王欢. 靶向干扰TAGLN表达对HBV阳性肝癌细胞生物学行为的影响及机制初探 ^*[J]. 中国生物工程杂志, 2019, 39(11): 13-21.

XU Yan,LIU Zheng-yun,ZHANG Wan-ling,WANG Sheng-yu,WANG Huan. Effect of Targeted Interference with TAGLN Expression on Biological Behavior of HBV-Positive Hepatocellular Carcinoma Cells and Its Mechanisms. China Biotechnology, 2019, 39(11): 13-21.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20191102 或 https://manu60.magtech.com.cn/biotech/CN/Y2019/V39/I11/13

图1 TAGLN在HBV阳性肝癌中的表达高于在HBV阴性肝癌中的表达

图2 shTAGLN1#-2.15和shTAGLN2#-2.15稳定表达细胞系中TAGLN的表达

图3 干扰TAGLN表达会抑制HepG2.2.15细胞增殖

图4 干扰TAGLN表达会抑制HepG2.2.15细胞迁移和侵袭

图5 HepG2.2.15细胞中PI3K和p-PI3K及AKT和p-AKT蛋白的表达

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