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目的:研究前列腺素E2(PGE2)对人肝细胞癌HepG2细胞E-cadherin表达的调节作用及其分子机制,探讨其与肝癌侵袭转移的关系,为肝癌的治疗提供新的研究思路。方法:COX-2瞬时转染人肝细胞癌HepG2细胞,Westernblot实验检测其E-cadherin表达的变化;PGE2及PI-3K抑制剂LY294002处理人肝细胞癌HepG2细胞,应用Westernblot和RT-PCR分别测定肝癌细胞内E-cadherin蛋白水平和mRNA水平的变化,并观察60min内Akt磷酸化水平的变化。结果:COX-2过表达转染HepG2细胞后,E-cadherin的表达明显降低;5、10、20μmol/LPGE2处理HepG2细胞24h后,E-cadherin表达水平与对照组比较分别下降了22.31%、36.3%、47.83%(P<0.05);20μmol/LPGE2处理HepG2细胞24、48、72h,E-cadherin蛋白和mRNA的表达与对照组比均明显降低(P<0.01)。PI-3K抑制剂LY294002能部分阻断PGE2对E-cadherin表达的调节作用;经PGE2处理的HepG2细胞Akt磷酸化水平显著升高,并且在15min时达到最高。结论:PGE2可以抑制人肝细胞癌HepG2细胞中E-cadherin的表达,并很可能通过激活Akt信号转导通路而发挥作用。
AIM: To investigate the regulatory effect of prostaglandin E2 (PGE2) on E-cadherin expression and its molecular mechanism in HepG2 human hepatocellular carcinoma (HCC), and to explore its relationship with invasion and metastasis of HCC and to provide a new research approach for the treatment of HCC. Methods: Human hepatocellular carcinoma HepG2 cells were transiently transfected with COX-2 and the expression of E-cadherin was detected by Western blot assay. Western blot and RT-PCR were used to detect the expression of E-cadherin in PGE2 and PI3K inhibitor LY294002 cells respectively E-cadherin protein level and mRNA level in hepatocellular carcinoma cells, and observe the change of Akt phosphorylation in 60min. Results: The expression of E-cadherin in HepG2 cells was significantly decreased after COX-2 overexpression. Compared with the control group, the expression of E-cadherin in HepG2 cells treated with 5, 10, 20 μmol / LPGE2 decreased by 22.31%, 36.3 %, 47.83% (P <0.05). The expression of E-cadherin protein and mRNA in HepG2 cells treated with 20μmol / LPGE2 for 24,48,72h were significantly lower than that of the control group (P <0.01). The PI-3K inhibitor LY294002 partially blocked the regulatory effect of PGE2 on E-cadherin expression. The phosphorylation of Akt in PGE2-treated HepG2 cells was significantly increased and reached its peak at 15 min. CONCLUSION: PGE2 can inhibit the expression of E-cadherin in HepG2 human hepatocellular carcinoma cells and may play a role by activating Akt signal transduction pathway.