m2型丙酮酸激酶促进肝癌细胞的转移作者董天赋柴浩陈圣林成峰.docVIP

M2型丙酮酸激酶促进肝癌细胞的转移 作 者：董天赋 柴浩 陈圣林 成　峰* * 通信作者（Corresponding authorE-mail：docchengfeng@ 位：南京医科大学第一附属医院肝脏移植中心,江苏 南京 210029;南京医科大学第一附属医院肝脏移植中心,江苏 南京 210029;南京医科大学第一附属医院肝脏移植中心,江苏 南京 210029;南京医科大学第一附属医院肝脏移植中心,江苏 南京 210029 基金项目：国家自然基金面上项目;省卫生厅重点项目(H201102);卫生厅开放课题(Z;省六大人才高峰项目(2009) 摘 要：目的:探讨PKM2（Pyruvate kinase M2）在肝癌组织中的表达及在肝癌侵袭中作用及相关机制。方法: 荧光定量PCR（Real-time PCR）, 免疫组化（IHC）比较PKM2在原发性肝癌和癌旁组织中mRNA和蛋白的表达水平；构建人类肝癌细胞株（HepG2）PKM2干扰和过表达的细胞株，分析PKM2对肝癌细胞转移的作用；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、HIF-1α（Hypoxia inducible factor-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，标本免疫组化（43/50）提示肝癌组织中的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的转移和侵袭，过表达PKM2促进肝癌细胞HepG2的转移和侵袭； 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的磷酸化，上调HIF-1α的表达，促进肝癌细胞的的转移和侵袭，为原发性肝癌的治疗提供了新思路。 [关键词]丙酮酸激酶M2型；原发性肝癌；肿瘤侵袭 Pyruvate kinase M2 Promotes Cell invasion in Liver Cancer Cell through Up-Regulating HIF-1α In Vitro Authors Name: Dong Tian-fu, Chai Hao,Chen Sheng-lin, Feng Cheng* Institution: Liver Transplantation Center, the First Affiliated Hospital of NJMU, Nanjing 210029,China;Liver Transplantation Center, the First Affiliated Hospital of NJMU, Nanjing 210029,China;Liver Transplantation Center, the First Affiliated Hospital of NJMU, Nanjing 210029,China;Liver Transplantation Center, the First Affiliated Hospital of NJMU, Nanjing 210029,China Abstract: Objective: To investigate the role of Pyruvate kinase M2（PKM2） for the invasion of liver cancer cell in vitro. Methods: Compared PKM2 expression in primary liver cancer tissues and adjacent normal tissues by Real-time PCR and immunohistochemistry. The lentivirus package system carrying PKM2/PKM-shRNA/Vector1/Vector2 was constructed, and HepG2 cells were infected by the virus. In addition, the invasion of HepG2 cells was observed after overexpressing or silencing. The expression of PKM2 and HIF-1α in liver cancer tissues was measured by Real-