维药异常黑胆质成熟剂对人肝癌HepG2细胞生物行为和Rho/ROCK信号通路的影响

 为探讨维药异常黑胆质成熟剂(ASM)对人肝癌细胞(HepG2)增殖、侵袭转移的影响及Rho/ROCK 信号传导通路相关蛋白表达影响,采用四甲基偶氮唑蓝(MMT)法检测不同浓度ASM(10、20、25、50 mg/mL)和10 μmol/L Y-27632 作用24、48、72 h后,对HepG2 细胞增殖的影响;采用扫描电镜技术和细胞侵袭实验测定ASM 不同剂量组和10 μmol/L Y-27632 作用24 h 癌细胞侵袭运动能力,Western Blot 检测ASM 不同剂量组和10 μmol/L Y-27632 作用24 h 癌细胞RhoA、ROCK1、ROCK2 的表达。结果显示,ASM 对肝癌细胞增殖有明显抑制作用,且表现为有明显的剂量效应关系:在10、20 mg/mL ASM 剂量组,ASM 药物作用24、48、72 h 后,HepG2 细胞增殖抑制作用随时间的延长而抑制增加,而25、50 mg/mL 剂量组,ASM 抑制细胞增殖作用不明显;ASM 抑制瘤细胞侵袭运动能力,扫描电镜结果显示ASM 抑制肿瘤细胞伪足的生长,ASM 中高剂量组ROCK1、ROCK2 的表达明显降低,RhoA 表达无明显变化。由此推论,ASM 对人肝癌细胞生长增殖和侵袭运动能力有抑制作用,其机制可能与ROCK酶表达降低有关。

Effect of Uyghur medicine ASM on cytobiology behavier and Rho/ROCK signal transduction pathway in human hepatoma HepG2 cells

This paper studies the effect of Uyghur medicine abnormal savda munzip (ASM) on the proliferation, invasion and the expression of Rho/ROCK signal transduction pathway related proteins in human hepatocellular carcinoma (HepG2) cells. After being treated with ASM (10, 20, 25, 50 mg/mL) and Y-27632(10 μmol/L) for 24, 48 and 72 h, the proliferation of HepG2 cells was detected using the four methyl thiazolyl tetrazolium (MTT) method. The alteration of the cells invasive ability treated with ASM and Y-27632 for 24h was observed by scanning electron microscope (SEM) and cell invasion assay. Western Blot was used to determine the effect of these drugs on the expression of RhoA, ROCK1 and ROCK2. The results revealed that ASM suppressed the proliferation of HepG2 significantly with a dose-effect relationship: for groups of ASM with 10 and 20 mg/mL, inhibition increased with the time of ASM action; for groups of ASM with 25 and 50 mg/mL, no significant increase of inhibition was found. ASM suppressed the invasive ability of HepG2. SEM results showed that the formation and growth of hepatoma cellular pseudopodia were inhibited by ASM. ASM with doses 25 and 50 mg/mL decreased the expressions of ROCK1 and ROCK2 significantly, but there was no significant effect on RhoA expression. The study suggests that ASM suppresses the proliferation and invasive ability of human hepatoma HepG2, the mechanisms of which could be associated with less expression of ROCK.