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

作　　者：周显飞 田舍[1] 王杰[1] 贾亮亮[1] 喻超[1] 江建新[1] 

机构地区：[1]贵州医科大学附院肝胆外科,贵州贵阳550004

出　　处：《贵阳医学院学报》2016年第9期997-1001,共5页Journal of Guiyang Medical College

基　　金：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)

摘　　要：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-29c on the biological characteristics of human pancreatic cancer cells （ AsPC-1 、BxPC-3 、PANC-1, MIA PaCa-2 ）. Methods： Culturing 1 nor- mal HPDE and 4 pancreatic cancer cells（AsPC-1 ,BxPC-3 、PANC-1 、MIA PaCa-2）. The different ex- pression of microRNA-29c in human pancreatic cancer lines were observed by real time RT-PCR. mi- croRNA-29c infected PANC-1 and MIA PaCa2 cells as experiment group, lip-null infected PANC-1 and MIA PaCa2 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-mes- enchymal transition （EMT） associated protein Vimentin and E-cadherin. Results： The result of real time RT-PCR showed that microRNA-29e expression in human pancreatic cancer cell lines were signifi- cantly 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 PaCa-2 cells in experiment group migrated obviously shorter than that of negative control group （P 〈 0.05 ）. TRanswell experiment discovered PANC-1 and MIA PaCa-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 PaCa-2 ceUs, Vimentin expression reduced and E-cadherin expression in- creased. Conclusion： The over-expression of microRNA-29c can effectively suppress the invasion me- tastasis human pancreatic cancer cells, indicating it might be a potential biological therapeutic target for pancreatic cancer.

关 键 词：微小RNA microRNA-29c 胰腺癌 腺病毒 细胞侵袭 转移 细胞划痕实验 

分 类 号：R735.9[医药卫生—肿瘤]