诱导性T细胞共刺激分子调控人肝癌HepG2细胞的增殖与侵袭。

Inducible T-cell co-stimulators regulate the proliferation and invasion of human hepatocellular carcinoma HepG2 cells.

作者信息

Wei Yaning, Wang Yanan, Zang Aimin, Shang Yanhong, Song Zizheng, Wang Zhiyu, Wang Yangyang, Yang Hua

机构信息

Departments of Oncology, Affiliated Hospital of Hebei University, Baoding, 071000, China.

Department of Pathology, Affiliated Hospital of Hebei University, Baoding, 071000, China.

出版信息

Biol Res. 2018 Jan 9;51(1):2. doi: 10.1186/s40659-017-0150-7.

DOI:10.1186/s40659-017-0150-7

PMID:29316975

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5759367/

Abstract

BACKGROUND

This study determined the regulatory effects of inducible T-cell co-stimulators (ICOS) in human hepatocellular carcinoma HepG2 cells using a RNA interference (RNAi) technique.

METHODS

A RNAi technique was used to knockdown the expression of ICOS. ICOS expression after knockdown was detected as mRNA and protein levels by RT-PCR and Western blot, respectively. A MTT colorimetric assay was used to detect cell proliferation, and the Transwell assay was used to detect cell invasion. Western blot was carried out to detect the level of Bcl-2, AKT, and PI3K protein expression in different groups.

RESULTS

The proliferation of HepG2 cells were significantly decreased after ICOS siRNA transfection (EG group). Similarly, the results of the Transwell experiment showed that invasion of HepG2 cells in the EG group was clearly reduced compared to the negative control (NC) and blank control groups (CON). Western blot analysis showed that knockdown of ICOS expression reduced the levels of Bcl-2 and AKT, and also significantly up-regulated the level of PI3K phosphorylation (P < 0.01).

CONCLUSION

Down-regulating ICOS expression in HepG2 cells suppressed cell proliferation and invasion. The underlying mechanism may be related to the expression of the downstream factor, PI3K/AKT.

摘要

背景

本研究采用RNA干扰（RNAi）技术，确定诱导性T细胞共刺激分子（ICOS）在人肝癌HepG2细胞中的调控作用。

方法

采用RNAi技术敲低ICOS的表达。分别通过RT-PCR和蛋白质免疫印迹法，在mRNA和蛋白质水平检测敲低后ICOS的表达。采用MTT比色法检测细胞增殖，采用Transwell实验检测细胞侵袭。通过蛋白质免疫印迹法检测不同组中Bcl-2、AKT和PI3K蛋白表达水平。

结果

ICOS siRNA转染后（EG组），HepG2细胞的增殖显著降低。同样，Transwell实验结果显示，与阴性对照组（NC）和空白对照组（CON）相比，EG组中HepG2细胞的侵袭明显减少。蛋白质免疫印迹分析显示，敲低ICOS表达降低了Bcl-2和AKT的水平，同时显著上调了PI3K磷酸化水平（P < 0.01）。