自分泌运动因子在肝细胞癌侵袭及转移中的作用

The Effect of Autocrine Motility Factor on the Invasion and Metastasis of Hepatocellular Carcinoma

目的:探讨自分泌运动因子(AMF)在人肝细胞癌侵袭和转移中的作用。方法:人肝细胞系LO2和人肝细胞癌细胞株MHCC97-H作为实验材料,检测二者AMF的表达水平;设计并合成针对AMF基因序列的双链小干扰RNA转染高转移性人肝癌细胞株MHCC97-H,Western blot检测AMF基因的蛋白的表达水平;通过MTT实验检测转染后细胞的增殖力;通过体外Transwell小室对比沉默AMF基因前后的肝癌细胞的迁移力和侵袭力;最后用细胞悬液皮下接种小鼠,观察沉默AMF基因前后肝细胞的成瘤能力。结果:AMF在MHCC97-H的表达量较高;将双链小干扰RNA转入MHCC97-H后,AMF的表达显著降低(P<0.05);沉默AMF基因序列后,MHCC97-H的增殖力、迁移力和侵袭力均有明显下降(P<0.05);用细胞悬液皮下接种小鼠沉默AMF基因的MHCC97-H形成的肿瘤体积小于对照组(P<0.05)。结论:AMF基因可调节肝癌细胞的迁移和侵袭。

Objective： To investigate the effect of autocrine motility factor invasion and metastasis of hepatocellular carcinoma.Methods： Human liver LO2 cells and human liver cancer MHCC97-H cells were used as the research object, and the protein expression levels of AMF was tested. Chemically synthesized si RNA duplex targeting the AMF gene（AMF-si RNA） was transiently transfected into MHCC97-H cells protein expression levels of AMF was detected by Western blot; Cell proliferation was determined by MTT experiment tests after transfection. The effect of AMF knockdown on cell invasion and migration was analyzed by Transwell chamber assays; With cell suspension finally subcutaneously vaccinated mice and observe silence before and after the AMF genetic liver cell into a tumor ability.Results： The protein expression of AMF in MHCC97-H quantity was higher; Putting the small double-stranded SiRNA transfection into MHCC97-H, the protein expression of AMF significantly reduced（P〈0.05）; After silence, AMF genetic sequences, MHCC97-H proliferation, migration and invasion forces were significantly decreased（P〈0.05）; Using cell suspension subcutaneous inoculation of AMF gene in mice MHCC97-H formation of tumor size was smaller than the control group（P〈0.05）. Conclusions： SiRNA-mediated silencing of AMF gene could significantly inhibit the cell invasion and metastasis of human hepatocellular carcinoma.