论文部分内容阅读
目的:分析肝细胞癌组织及细胞系中脂肪甘油三酯脂肪酶(adipose triglyceride lipase,ATGL)的表达水平,探讨ATGL在肝细胞癌细胞增殖、迁移和侵袭过程中的作用。方法:采用蛋白质印迹法检测57对肝细胞癌组织和相应癌旁正常肝组织,以及12种肝癌细胞系中ATGL的表达水平。将靶向ATGL基因的shRNA重组慢病毒(GV248-shATGL-1、-2和-3)及其阴性对照慢病毒(GV248-shNC)分别感染肝癌Huh7和MHCC-97L细胞系,然后采用实时荧光定量PCR和蛋白质印迹法验证ATGL基因沉默效率。采用CCK-8法和Transwell小室法分别检测ATGL基因沉默对肝癌Huh7和MHCC-97L细胞增殖、迁移和侵袭的影响。结果:57例肝细胞癌患者中,有28例(49%)肝癌组织的ATGL表达水平明显高于其相应癌旁肝组织,16例(28%)肝癌组织的ATGL表达水平低于相应癌旁肝组织,而13例(23%)无明显变化。与阴性对照组相比,沉默ATGL基因表达能够明显抑制肝癌Huh7和MHCC-97L细胞增殖(P值均<0.05),同时细胞迁移和侵袭能力均明显降低(P值均<0.01)。结论:肝细胞癌组织中ATGL相对高表达,而沉默ATGL基因表达可以抑制肝细胞癌Huh7和MHCC-97L细胞的增殖、迁移和侵袭。
OBJECTIVE: To analyze the expression of adipose triglyceride lipase (ATGL) in hepatocellular carcinoma tissues and cell lines and to explore the role of ATGL in the proliferation, migration and invasion of hepatocellular carcinoma cells. Methods: Western blotting was used to detect the expression of ATGL in 57 hepatocellular carcinoma tissues, corresponding normal liver tissues and 12 hepatocellular carcinoma cell lines. The shRNA recombinant lentivirus (GV248-shATGL-1, -2 and -3) and its negative control lentivirus (GV248-shNC) targeting the ATGL gene were respectively transfected into Huh7 and MHCC-97L cell lines, Validation of ATGL Gene Silencing Efficiency by PCR and Western Blotting. The effects of ATGL gene silencing on proliferation, migration and invasion of HCC Huh7 and MHCC-97L cells were detected by CCK-8 assay and Transwell chamber assay. Results: The ATGL expression in hepatocellular carcinoma was significantly higher in 28 cases (49%) of 57 hepatocellular carcinoma patients than in its corresponding paracancer liver tissue. The ATGL expression in 16 cases (28%) of liver cancer tissues was lower than that in corresponding paracancer Liver tissue, while 13 cases (23%) had no significant change. Compared with the negative control group, silencing ATGL gene expression significantly inhibited the proliferation of Huh7 and MHCC-97L cells (all P <0.05), and significantly decreased the ability of cell migration and invasion (P <0.01). CONCLUSION: ATGL is relatively highly expressed in hepatocellular carcinoma, while silencing ATGL gene expression can inhibit the proliferation, migration and invasion of hepatocellular carcinoma Huh7 and MHCC-97L cells.