M2-型丙酮酸激酶促进肝癌细胞的侵袭
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国家自然科学基金面上项目(81070324);江苏省卫生厅重点项目(H201102);卫生厅开放课题(ZX05200906);江苏省六大人才高峰项目(2009)


Promotion of pyruvate kinase M2 on cell invasion in liver cancer cell
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    摘要:

    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-

    Abstract:

    Objective:To investigate the role of pyruvate kinase M2 (PKM2) on the invasion of liver cancer cell in vitro. Methods:We compared mRNA and protein expressions of PKM2 in hepatocellular carcinoma(HCC) tissues and adjacent normal tissues by real-time PCR and immunohistochemistry. Interfered and overexpressed PKM2 cell lines of HepG2 were constructed to analyze the role of PKM2 on invasion of liver cancer cells. Real-time PCR,immunohistochemistry and Western blot were performed to investigate the expression of HIF-1α and the phosphorylation of STAT3 in HepG2 cells. Results:The mRNA expressions of PKM2 and HIF-1α in the HCC tissues(106/115) were higher than those in adjacent normal tissues. Sample immunohistochemistry showed that the expressions of PKM2 and HIF-1α in HCC tissues were higher than those in adjacent normal tissues. In vitro experiment,interference of PKM2 inhibited the invasion of HepG2 liver cancer cell,and overexpression of PKM2 promoted the invasion of HepG2. The overexpression of PKM2 promoted the phosphorylation of signal transducer and activator of transcription 3(STAT3) and upregulated the expression of HIF-1α. Conclusion:PKM2 upregulates the expression of HIF-1α and promotes liver cancer cell invasion by promoting the phosphorylation of STAT3 and may represent a novel strategy for therapy of HCC.

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董天赋,柴浩,陈圣林,熊新魁,孙道一,成峰. M2-型丙酮酸激酶促进肝癌细胞的侵袭[J].南京医科大学学报(自然科学版),2014,(4):410-416

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  • 收稿日期:2013-12-18
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  • 在线发布日期: 2014-04-18
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