Sun Xiangjun, Wang Mingchun, Zhang Fenghua, Kong Xiao
Department of Hepatobiliary Surgery, Linyi People's Hospital, Linyi, Shandong 276003, P.R. China.
Department of Surgery, Linyi People's Hospital, Linyi, Shandong 276003, P.R. China.
Exp Ther Med. 2019 Mar;17(3):2334-2340. doi: 10.3892/etm.2019.7211. Epub 2019 Jan 29.
The present study aimed to elucidate the underlying mechanism of neuroepithelial cell transforming 1 (NET-1), a member of the Ras homolog gene family, in hepatocellular carcinoma (HCC). To determine the association between the expression of NET-1 and the proliferation and migration of MHCC97-H cells, the cells were transfected with NET-1 small interfering (si)RNA and si negative control. Following transfection with NET-1 siRNA, the proliferation rate of MHCC97-H cells decreased significantly and the percentage of apoptotic cells increased. The HCC cell line MHCC97-H was used in the present study as it exhibited an increased expression level of NET-1 compared with the MHCC97-L cell line. Expression levels of apoptosis-associated proteins including apoptosis regulator Bax (Bax), cyclinD1, apoptosis regulator Bcl-2 (Bcl-2) and caspase-3 were determined. Expression levels of phosphoinositide 3-kinase (PI3K) and protein kinase B (AKT) and their phosphorylated forms were also measured by western blotting. Following NET-1 knockdown, the expression of Bax and cyclinD1 decreased, the expression of Bcl-2 and caspase-3 increased, and the PI3K/AKT signaling pathway was inhibited. The results of the present study suggest that inhibition of NET-1 can suppress the progression of HCC by targeting the PI3K/AKT signaling pathway. NET-1 expression level in HCC cells increased compared with normal liver cells.
本研究旨在阐明Ras同源基因家族成员神经上皮细胞转化1（NET-1）在肝细胞癌（HCC）中的潜在机制。为了确定NET-1的表达与MHCC97-H细胞增殖和迁移之间的关联，将细胞用NET-1小干扰（si）RNA和si阴性对照进行转染。用NET-1 siRNA转染后，MHCC97-H细胞的增殖率显著降低，凋亡细胞百分比增加。本研究使用HCC细胞系MHCC97-H，因为与MHCC97-L细胞系相比，它表现出NET-1表达水平升高。测定了凋亡相关蛋白的表达水平，包括凋亡调节因子Bax（Bax）、细胞周期蛋白D1、凋亡调节因子Bcl-2（Bcl-2）和半胱天冬酶-3。还通过蛋白质印迹法测量了磷酸肌醇3激酶（PI3K）和蛋白激酶B（AKT）及其磷酸化形式的表达水平。NET-1敲低后，Bax和细胞周期蛋白D1的表达降低，Bcl-2和半胱天冬酶-3的表达增加，PI3K/AKT信号通路受到抑制。本研究结果表明，抑制NET-1可通过靶向PI3K/AKT信号通路抑制HCC的进展。与正常肝细胞相比，HCC细胞中NET-1表达水平升高。
