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目的探讨肝星状细胞(HSC)对肝癌细胞(HCC)恶性生物学行为的影响及其相关机制。方法分别培养SMMC-7721肝癌细胞、Hep G2肝癌细胞和LX-2 HSC,采用LX-2 HSC条件培养基(LX2-CM)、丝裂原激活蛋白激酶(MAPK)特异性抑制剂U0126处理肝癌细胞,TranswellTM小室检测肝癌细胞的侵袭和迁移能力,CCK-8法检测细胞的增殖情况,实时定量PCR和Western blot法分别检测两种肝癌细胞磷酸化的胞外信号调节激酶1/2(p-ERK1/2)、ERK1/2、c-Myc、波形蛋白(vimentin)、上皮钙黏素(E-cadherin)的mRNA和蛋白水平。结果 LX2-CM能促进SMMC-7721细胞和Hep G2细胞的增殖、侵袭和迁移,其作用可被U0126阻断。LX2-CM可以上调p-ERK1/2、c-Myc、vimentin的水平,下调E-cadherin的水平;U0126处理细胞后,p-ERK1/2、c-Myc、vimentin的水平显著降低,E-cadherin水平明显升高。结论 LX2-CM能通过ERK1/2通路激活c-Myc,促进肝癌细胞的增殖、侵袭和迁移,并诱导上皮间质转化。
Objective To investigate the effect of hepatic stellate cells (HSC) on the malignant biological behavior of hepatocellular carcinoma (HCC) and its related mechanisms. METHODS: SMMC-7721 hepatocarcinoma cells, Hep G2 hepatocarcinoma cells and LX-2 HSCs were cultured with LX-2 HSC conditioned medium (LX2-CM) and mitogen-activated protein kinase (MAPK) specific inhibitor U0126 TranswellTM chamber was used to detect the invasion and migration of hepatocellular carcinoma cells. The proliferation of hepatocellular carcinoma cells was detected by CCK-8 assay. The phosphorylation of extracellular signal-regulated kinase 1/2 (p-ERK1) was detected by real-time quantitative PCR and Western blot, respectively / 2), ERK1 / 2, c-Myc, vimentin, E-cadherin mRNA and protein levels. Results LX2-CM can promote the proliferation, invasion and migration of SMMC-7721 cells and Hep G2 cells, and its effect can be blocked by U0126. LX2-CM can upregulate the expression of p-ERK1 / 2, c-Myc and vimentin and down-regulate the expression of E-cadherin. The levels of p-ERK1 / 2, c-Myc and vimentin in U0126- The level was significantly higher. Conclusion LX2-CM can activate c-Myc through ERK1 / 2 pathway, promote the proliferation, invasion and migration of hepatocellular carcinoma cells and induce epithelial-mesenchymal transition.