人工高表达LAIR-1对人肝癌细胞系SMMC-7721细胞增殖、凋亡和侵袭的影响

The effect of LAIR-1 Overexpression on proliferation,apoptosis and invasion of hepatoma cell line SMMC-7721

研究人白细胞相关免疫球蛋白样受体1(leukocyte-associated immunoglobulin-like receptor 1,LAIR-1)对人肝癌细胞系SMMC-7721细胞增殖、凋亡和侵袭的影响,探讨LAIR-1的表达在肝癌中的作用。以LAIR-1慢病毒表达载体(LV-LAIR-1)转染SMMC-7721细胞,建立稳定高表达LAIR-1的细胞株LAIR-1+SMMC-7721。采用流式细胞术(flow cytometry,FCM)、Western blotting、RT-PCR和激光扫描共聚焦显微技术(laser scanning confocal microscopy,LSCM)等方法鉴定LAIR-1分子在SMMC-7721细胞的表达水平;采用CCK-8试剂盒、FITC Annexin V凋亡试剂盒和Transwell侵袭实验分别检测LAIR-1对SMMC-7721细胞增殖、凋亡和侵袭的影响。检测结果显示,经LV-LAIR-1转染的SMMC-7721细胞高水平表达LAIR-1分子;功能实验结果显示,与实验组细胞相比,LAIR-1的表达对细胞增殖无明显影响(P>0.05),但是能够明显促进细胞凋亡(P<0.05),并能够显著抑制细胞的侵袭能力(P<0.05)。上述研究结果提示,LAIR-1分子的表达可能是影响肝癌细胞生物学特性的一个重要因素。

To investigate the effects of the expression of leukocyte-associated immunoglobulin-like receptor -1 （LAIR-1） on the proliferation, apoptosis and invasion of hepatoma cell line SMMC-7721, and to explore the role of LAIR-1 in the hepatocarcinoma, we introduced LAIR-1 gene into SMMC 7721 cells by LAIR-1 lentiviral vector（LV-LAIR-1）to establish a cell line with high and stable expression of LAIR -1 （LAIR-1＋ SMMC-7721）. Flow cytometry, Western blotting, real-time PCR and laser scanning confocal microscopy were used to measure the level of LAIR-1 in SMMC-7721 cells. The influence of LAIR- 1 on the proliferation, apoptosis and invasion of SMMC-7721 cells was detected by CCK-8 kit, FITC Annexin V apoptosis detection kit and transwell assay respectively. The results indicated that the expression of LAIR 1 at both mRNA and protein levels signifi cantly increased in SMMC 7721 cells after stable gene transfection with LV-LAIR-1. Results of functional experiments showed that compared with the control, high expression of LAIR-1 significantly promoted the apoptosis（P 〈0.05） and inhibited the invasions（P 〈0.05） of SMMC-7721 cells, but no significant influence on cell proliferation（P 〉0.05） was detected. In conclu sion, LAIR-1 may play a role in the biological characteristics of hepatocarcinoma.