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目的:探讨M2-型丙酮酸激酶(pyruvate kinase M2,PKM2)在肝癌组织中的表达及对肝癌侵袭的影响。方法:荧光定量PCR(real-time PCR)、免疫组化(IHC)比较PKM2在肝癌和癌旁组织中mRNA和蛋白的表达水平;构建人肝癌细胞株(HepG2)PKM2干扰和过表达的细胞株,分析PKM2对肝癌细胞侵袭的影响;Real-time PCR、免疫组化、Western blot分析肝癌组织及转染后的HepG2细胞STAT3的磷酸化、缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)的表达变化。结果:肝癌标本(106/115)中PKM2、HIF-1α的mRNA表达水平在肝癌组织中高于癌旁组织,免疫组化显示肝癌组织中的PKM2、HIF-1α的表达水平高于癌旁组织;在体外实验中,干扰PKM2能抑制肝癌细胞HepG2的侵袭,过表达PKM2促进肝癌细胞HepG2的侵袭;PKM2过表达促进STAT3(signal transducer and activator of transcription 3)的磷酸化,上调HIF-1α的表达。结论:PKM2通过促进STAT3磷酸化,上调HIF-1α的表达,促进肝癌细胞的侵袭,为肝癌的治疗提供了新思路。
Objective: To investigate the expression of pyruvate kinase M2 (PKM2) in hepatocellular carcinoma (HCC) and its effect on the invasion of liver cancer. Methods: The mRNA and protein expressions of PKM2 were detected by real-time PCR and immunohistochemistry (IHC) in human hepatocellular carcinoma (HCC) and adjacent non-cancerous tissues. The constructed cell lines with interference and over-expression of PKM2 by human hepatoma cell line HepG2 The effect of PKM2 on the invasion of hepatocellular carcinoma cells was analyzed. The expressions of phosphorylated STAT3 and hypoxia inducible factor-1α (HIF-1α) in HepG2 cells were detected by Real-time PCR, immunohistochemistry and Western blot. -1α) expression changes. Results: The mRNA expression of PKM2 and HIF-1α in HCC tissues was higher than that in paracancerous tissues in HCC tissues. The expression of PKM2 and HIF-1α in HCC tissues was higher than that in paracancerous tissues. In vitro, PKM2 could inhibit the invasion of HepG2 cells and over-expression of PKM2 could promote the invasion of HepG2 cells. PKM2 overexpression promoted the phosphorylation of signal transducer and activator of transcription 3 and up-regulated the expression of HIF-1α. Conclusion: PKM2 can promote the invasion of hepatocellular carcinoma cells by promoting the phosphorylation of STAT3, increasing the expression of HIF-1α and providing a new idea for the treatment of hepatocellular carcinoma.