[TSLC1基因对人肝癌细胞系HepG2的生长抑制作用]

[The growth inhibition effects of TSLC1 gene on human hepatocyte carcinoma cell line HepG2].

作者信息

Qin Li, Zhang Zheng-mao, Hao You-hua, Wang Bao-ju, Yang Xin-xing, Tian Yong-jun, Yang Dong-liang

机构信息

Division of Clinical Immunology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2007 Jul;15(7):509-12.

PMID:17669239

Abstract

OBJECTIVES

To study the effects of tumor suppressor in lung cancer-1 (TSLC1) on human hepatocyte carcinoma cell line HepG2.

METHODS

A full length of TSLC1 cDNA was amplified from RNA of normal human liver cells by RT-PCR, and it was cloned into a pCI-neo expression vector and transfected into human hepatocellular carcinoma cell line HepG2. The HepG2 cells transfected with this plasmid (experimental group) and those treated with pCI-neo vector (control group) and without any treatment (blank group) were compared. Cell morphology was studied microscopically and cell growth was analyzed with MTT assay. FACSort flow cytometry analysis was performed to assess the cell cycle distribution and apoptosis.

RESULTS

A stable cell line expressing TSLC1 protein was successfully established. Morphologically, cells of the experimental group were tightly aggregated when compared with those of the control and blank groups. The growth of TSLC1-transfected cells was significantly suppressed in vitro compared with those of the control and blank groups. The amount of G0-G1 cells was 63.66%+/-3.83% (P less than 0.01) in the experimental group, while those of the control and blank groups were 47.45%+/-0.91% and 54.47%+/-0.96% respectively. The amount of S phase cells in the experimental group, 22.90%+/-6.04%, was significantly lower (P less than 0.05) than that of the control group (36.58%+/-0.61%) and the blank group (33.61%+/-2.99%), which suggested a G0-G1 cell cycle arresting. The number of cells in early and late phase apoptosis (17.09%+/-0.20% and 16.11%+/-0.40% respectively) were significantly higher than those of the control and blank groups (P less than 0.01).

CONCLUSIONS

TSLC1 strongly inhibits the growth of HepG2 cells in vitro and induces apoptosis of the cells, suggesting that TSLC1 may have a tumor suppressor function in HCC.

摘要

目的

研究肺癌抑癌基因1（TSLC1）对人肝癌细胞系HepG2的影响。

方法

通过逆转录聚合酶链反应（RT-PCR）从正常人肝细胞RNA中扩增出TSLC1全长cDNA，将其克隆到pCI-neo表达载体中并转染到人肝癌细胞系HepG2。比较转染该质粒的HepG2细胞（实验组）、用pCI-neo载体处理的细胞（对照组）和未作任何处理的细胞（空白组）。通过显微镜观察细胞形态，并用MTT法分析细胞生长情况。采用流式细胞术分析细胞周期分布及凋亡情况。

结果

成功建立了表达TSLC1蛋白的稳定细胞系。形态学上，与对照组和空白组相比，实验组细胞紧密聚集。与对照组和空白组相比，转染TSLC1的细胞在体外生长明显受到抑制。实验组G0-G1期细胞比例为63.66%±3.83%（P<0.01），而对照组和空白组分别为47.45%±0.91%和54.47%±0.96%。实验组S期细胞比例为22.90%±6.04%，明显低于对照组（36.58%±0.61%）和空白组（33.61%±2.99%）（P<0.05），提示细胞周期阻滞于G0-G1期。早期和晚期凋亡细胞数量（分别为17.09%±0.20%和16.11%±0.40%）明显高于对照组和空白组（P<0.01）。