摘要
目的阐明单酰甘油脂肪酶(monoacylglycerol lipase, MAGL)对肝细胞癌(hepatocellular carcinoma, HCC)细胞株增殖、凋亡与侵袭的影响及其机制。方法比较多种HCC细胞株(SMMC-7721、HepG2、Huh7. 0)和正常肝细胞株(L02)以及同源但不同侵袭能力的HCC细胞株(HCCM3、HCCM6、MHCC-97H、MHCC-97L)中MAGL蛋白表达差异。硅酸盐纳米介孔材料LPSS-NH2介导的RNAi(MAGL-shRNA)、过表达和特异性药物JZL-184调控MAGL表达与功能,观察多种HCC细胞株和正常肝细胞株的增殖、凋亡和侵袭能力变化。ELISA法检测干预MAGL后其可能的下游信号分子前列腺素E2 (prostaglandin E2, PGE2)和溶血性磷脂酸(lysophosphatidic acid, LPA)水平变化。结果 MAGL蛋白表达水平在HCC细胞株中明显高于正常肝细胞株L02 (P <0. 05),且高侵袭力的HCC细胞株MAGL表达高于同源但低侵袭力的HCC细胞株(P <0. 05)。下调MAGL的表达可抑制HCC细胞株的增殖,促进其凋亡,并抑制其侵袭能力,过表达则反之,且过表达MAGL诱导正常肝细胞株L02产生了侵袭能力。PGE2随着MAGL的表达调控呈正相关变化,而LPA则无明显变化趋势。结论 LPSSNH2介导的MAGL-shRNA转染肝细胞癌细胞株,可通过抑制MAGL的高表达并下调其下游PGE2水平,抑制肝细胞癌细胞增殖与侵袭。
Objective To determine the effects of monoacylglycerol lipase （MAGL） on proliferation, apoptosis and invasion of hepatocellular carcinoma （HCC） cell lines and investigate the potential mechanisms. Methods The protein level of MAGL was detected in various HCC cell lines （SMMC-7721, HepG2, Huh7.0）, normal liver cell line （L02） and HCC cell lines （HCCM3, HCCM6, MHCC-97H and MHCC-97L） with different invasion abilities. With the aid of silicate nano mesoporous materials （LPSS-NH2）, MAGL-iRNA, over-expression plasmid Overexp-MAGL and specific inhibitor of MAGL JZL-184 were respectively transfected to silence or inhibit the expression and function of MAGL in SMMC-7721, HepG2, Huh7.0 and L02 cells. Then the proliferation, apoptosis and invasion ability were measured by CCK-8 assay, flow cytometry and Transwell chamber test, respectively. ELISA was adopted to detect the supernatant and intracellular contents of potential down-stream signal molecules prostaglandin E2 （PGE2） and lysophosphatidic acid （LPA）. ResultsThe protein level of MAGL was significant higher in the HCC cell lines than L02 cells （P〈0.05）, and in the cell lines with higher invasion ability than those with lower （P〈0.05）. Down-regulation of MAGL inhibited cell proliferation, promoted cell apoptosis and suppressed invasion ability. While its over-expression presented opposite results, but induced invasion ability in L02 cells. The supernatant and intracellular contents of PGE2 were in concordance with the regulation of MAGL, but no such changes were found in the content of LPA. Conclusion LPSS-NH2 mediated MAGL-shRNA transfection down-regulates PGE2 and then inhibits proliferation and invasion in hepatocellular cells.
作者
陈先锋
张俊勇
许仲平
龚建平
隆洪木
CHEN Xianfeng1, ZHANG Junyong2 , XU Zhongping1 , GONG Jianping2, LONG Hongmu1(1 Department of Hepatobiliary Surgery, Fuling Center Hospital, Chongqing, 408000 ; 2Department of Hepatobiliary Surgery, the Second Hospital of Chongqing Medical University, Chongqing, 400010, China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2018年第19期1762-1769,共8页
Journal of Third Military Medical University
基金
重庆市卫生和计划生育委员会科研项目(zdxk201605)~~
