| PKM2基因对胆管细胞癌迁移､侵袭及增殖的影响 |
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| 引用本文： | 柴 浩,熊新魁,孙道一,单文刚,浦立勇,俞 悦,成 峰. PKM2基因对胆管细胞癌迁移､侵袭及增殖的影响[J]. 南京医科大学学报(自然科学版), 2015, 0(5): 615-621 |
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| 作者姓名： | 柴 浩 熊新魁 孙道一 单文刚 浦立勇 俞 悦 成 峰 |
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| 作者单位： | 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,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 |
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| 基金项目： | 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) |
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| 摘 要： | 目的:检测M2-型丙酮酸激酶(pyruvate kinase M2,PKM2)在胆管癌(cholangiocarcinoma,CCA)组织中的表达情况,探讨PKM2下调对胆管癌细胞迁移?侵袭及增殖的影响?方法:实时定量逆转录聚合酶链反应(qRT-PCR)和免疫组织化学(immunohistochemistry,IHC)染色分别检测胆管癌及对应癌旁组织标本中PKM2的mRNA和蛋白表达水平?利用慢病毒表达载体系统在胆管癌细胞株HuCCT-1?HCCC-9810中下调PKM2,分别用划痕实验?Transwell细胞侵袭实验和CCK-8比色法检测细胞迁移?侵袭及增殖能力?结果:胆管癌组织中PKM2的mRNA和蛋白表达水平明显高于对应癌旁组织?经qRT-PCR和Western blot方法证实稳定转染PKM2 shRNA的胆管癌细胞中PKM2的mRNA和蛋白水平较对照组均明显下降(P < 0.05),与空载对照组(PKM2-NC)和正常对照组(PKM2)相比,实验组细胞(PKM2-shRNA)的迁移?侵袭及增殖能力明显减弱,差异有统计学意义(P < 0.05)?结论:胆管癌组织中PKM2表达明显高于癌旁组织,PKM2 shRNA能有效地降低胆管癌细胞中PKM2基因的表达,PKM2基因沉默可以抑制HuCCT-1?HCCC-9810细胞的迁移?侵袭及增殖?
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| 关 键 词： | shRNA干扰 PKM2基因 生物学功能 |
| 收稿时间： | 2015-01-13 |
Au experimental research on PKM2 gene on migration, invasion and proliferation of cholangiocarcinoma cell line |
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| Chai Hao,Xiong Xinkui,Sun Daoyi,Shan Wengang,Pu Liyong,Yu Yue and Cheng Feng. Au experimental research on PKM2 gene on migration, invasion and proliferation of cholangiocarcinoma cell line[J]. Acta Universitatis Medicinalis Nanjing, 2015, 0(5): 615-621 |
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| Authors: | Chai Hao Xiong Xinkui Sun Daoyi Shan Wengang Pu Liyong Yu Yue Cheng Feng |
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| Affiliation: | 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,Liver Transplantation Center ,the First Affiliated Hospital of NJMU,Nanjing 210029,China,Liver Transplantation Center ,the First Affiliated Hospital of NJMU,Nanjing 210029,China and Liver Transplantation Center ,the First Affiliated Hospital of NJMU,Nanjing 210029,China |
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| Abstract: | Objective:To investigate the expression level of PKM2 in cholangiocarcinoma (CCA)tissues,then study the effect of PKM2 down-regulation on migration,invasion and proliferation in cholangiocarcinoma cell lines. Methods:RNA and protein expressions of PKM2 in CCA tissues and paired adjacent tissues were detected by qRT-PCR and immunohistochemistry. PKM2 was down-regulated by a lentiviral vector expression system in cholangiocarcinoma cell lines HuCCT-1 and HCCC-9810. qRT-PCR and Western blot were performed to analyze the mRNA and protein expression of PKM2 in both cell lines. Cell migration, invasion and proliferation were assessed by wound-healing experiment, matrigel invasion and Cell Counting Kit-8(CCK-8). Results:The expression of PKM2 in CCA tissues had a higher level than that in paired adjacent tissues. The mRNA and protein expressions of PKM2 in the experimental group (PKM2-shRNA)were significantly lower than those in the two control groups,confirmed by qRT-PCR and Westen blot (P < 0.05). Compared to the empty vector group (PKM2-NC)and the normal control group (PKM2),the cell invasion,migration and proliferation were significantly decreased in the experimental group (PKM2-shRNA)(P < 0.05). Conclusion:Down-regulation of PKM2 by PKM2 shRNA can inhibit migration,invasion and proliferation of HuCCT-1 and HCCC-9810 cells. |
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| Keywords: | shRNA interference PKM2 gene biological function |
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