贵州医科大学学报

2016, v.41;No.192(09) 997-1001

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microRNA-29c对人胰腺癌细胞侵袭转移的抑制作用及机制
Study on microRNA-29c Inhibition Effect and Mechanism on Human Pancreatic Cancer Cells Migration

周显飞;田舍;王杰;贾亮亮;喻超;江建新;
ZHOU Xianfei;TIAN She;WANG Jie;JIA Liangliang;YU Chao;JIANG Jianxin;Department of Hepatobiliary Surgery,the Affiliated Hospital of Guizhou Medical University;

摘要(Abstract):

目的:探讨microRNA-29c对人胰腺癌细胞(As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2)生物学特性的影响。方法:培养1种人正常胰腺上皮细胞(HPDE)及4种人胰腺癌细胞(As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2),采用real-time PCR法观察5种细胞系中microRNA-29c的表达差异,以microRNA-29c过表达腺病毒感染的PANC-1和MIA Pa Ca2细胞作为实验组,以空载体感染的PANC-1和MIA Pa Ca2细胞作为阴性对照组,采用细胞划痕实验、Transwell法检测两组细胞体外侵袭能力,Western blot检测两组细胞上皮间充质转化(EMT)相关蛋白波形蛋白(Vimentin)及E-钙粘蛋白(E-cadherin)的表达。结果:real-time-PCR显示各胰腺癌细胞系中microRNA-29c水平明显低于正常胰腺细胞系(P<0.05),细胞划痕实验发现感染腺病毒48 h后实验组PANC-1、MIA Pa Ca-2细胞的迁移距离明显短于阴性对照组(P<0.05),Transwell小室细胞侵袭实验发现实验组PANC-1和MIA Pa Ca-2细胞侵袭数量明显低于阴性对照组(P<0.05);Western blot蛋白免疫印迹结果显示PANC-1和MIA Pa Ca-2细胞过表达microRNA-29c后,Vimentin表达减少,E-cadherin表达增加。结论:microRNA-29c的过表达可有效抑制胰腺癌细胞的体外侵袭与转移,可能与Vimentin表达减少,E-cadherin表达增加有关,有望成为胰腺癌生物治疗的潜在靶点。
Objective: To investigate the effect of microRNA-29 c on the biological characteristics of human pancreatic cancer cells( As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2). Methods: Culturing 1 normal HPDE and 4 pancreatic cancer cells( As PC-1、Bx PC-3、PANC-1、MIA Pa Ca-2). The different expression of microRNA-29 c in human pancreatic cancer lines were observed by real time RT-PCR. microRNA-29 c infected PANC-1 and MIA Pa Ca2 cells as experiment group,lip-null infected PANC-1and MIA Pa Ca2 cells as negative control group; wound healing experiment,Transwell method to test the vitro invasion of both group cells; Western blot was used to measure the change of epithelial-mesenchymal transition( EMT) associated protein Vimentin and E-cadherin. Results: The result of real time RT-PCR showed that microRNA-29 c expression in human pancreatic cancer cell lines were significantly lower than that in the normal pancreatic ductal epithelial cells( HPDE)( P < 0. 05); wound healing experiment discovered that 48 hr after infection,PANC-1 and MIA Pa Ca-2 cells in experiment group migrated obviously shorter than that of negative control group( P < 0. 05). TRanswell experiment discovered PANC-1 and MIA Pa Ca-2 cells in experiment group invasion number was obviously shorter than that of negative control group( P < 0. 05). Western blot indicated after overexpression microRNA-29c of PANC-1 and MIA Pa Ca-2 cells,Vimentin expression reduced and E-cadherin expression increased. Conclusion: The over-expression of microRNA-29 c can effectively suppress the invasion metastasis human pancreatic cancer cells,indicating it might be a potential biological therapeutic target for pancreatic cancer.

关键词(KeyWords): 微小RNA;microRNA-29c;胰腺癌;腺病毒;细胞侵袭;转移;细胞划痕实验
micro RNA;microRNA-29c;pancreatic cancer;adenovirus;cell invasion;metastasis;wound healing experiment

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基金项目(Foundation): 国家国际科技合作专项资助(2014DFA31420);; 国家自然科学基金资助项目(81160311)

作者(Author): 周显飞;田舍;王杰;贾亮亮;喻超;江建新;
ZHOU Xianfei;TIAN She;WANG Jie;JIA Liangliang;YU Chao;JIANG Jianxin;Department of Hepatobiliary Surgery,the Affiliated Hospital of Guizhou Medical University;

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