miR-103a-3p在胰腺癌细胞中的表达及意义

摘要

Objective: To observe the expression inhibition of miR-103-a-3p in PANC-1 pancreatic cancer cell lines infected by lentivirus with miR-103a-3p-inhibitor, and to provide theoretical basis for elucidating the pathogenesis of pancreatic cancer and screening novel target genes for the treatment of .pancreatic cancer.Method:The expression of miR-103a-3p in two pancreatic cancer cell strains (PANC-1 and BxPC-3) with different invasiveness and the normal pancreatic ductal epithelial cell line (H6C7) were detected by using qRT-PCR. Lentivirus vectors with miR-103a-3p-inhibitor were constructed and used to infect PANC-1 cell with high expression level of miR-103a-3p. Transfection efficiency was observed by fluorescence microscope and the expression of miR-103a-3p were detected by qRT-PCR. Result:qRT-PCR showed that the relative quantitative expressions of miR-103a-3p in PANC-1, BxPC-3 and H6C7 cell strains were (4.949±0.130), (1.417±0.120) and (1.010±0.151), respectively. miR-103a-3p was expressed at highest level in PANC-1 cell. After infected by lentivirus with miR-103a-3p-inhibitor, PANC-1 cells were found to express green fluorescent proteins at high level. qRT-PCR indicated that there were no significant differences in the relative quantitative expressions of miR-103a-3p between PANC-1 cells uninfected by lentivirus with miR-103a-3p-inhibitor vector (1.002±0.160) and PANC-1 cells infected by lentivirus with negative control vector (0.956±0.250), but significant reduction of the relative quantitative expressions of miR-103a-3p in PANC-1 cells infected by lentivirus with miR-103a-3p-inhibitor vector (0.317±0.320) were detected(P<0.05).Conclusion:miR-103a-3p is expressed at higher level in pancreatic cancer cells with stronger invasion ability. miR-103a-3p-inhibitor can be transfected stablely into PANC-1 cells and inhibit the expression of miR-103a-3p.%目的：观察miR-103a-3p 抑制表达的慢病毒载体对高表达miR-103a-3p的PANC-1胰腺癌细胞株的表达抑制作用，为阐明胰腺癌的发病机制及筛选治疗靶基因提供理论依据。方法：采用实时荧光定量PCR（qRT-PCR）检测不同侵袭能力的胰腺癌细胞株（PANC-1、BxPC-3）和永生化正常胰腺细胞（H6C7）中miR-103a-3p表达情况；构建慢病毒载体miR-103a-3p-inhibitor，包装后感染高表达miR-103a-3p的PANC-1细胞株，荧光显微镜观察其转染效率，qRT-PCR检测其表达情况。结果：qRT-PCR检测显示，miR-103a-3p在PANC-1、BxPC-3细胞及正常胰腺细胞H6C7细胞中相对表达量分别为（4.949±0.130）、（1.417±0.120）和（1.010±0.151），PANC-1表达水平最高（P<0.05）。miR-103a-3p-inhibitor慢病毒感染 PANC-1细胞第4天，荧光显微镜下可见报告基因绿色荧光蛋白（GFP）高强度表达。qRT-PCR结果显示：miR-103a-3p相对表达量在未感染miR-103a-3p-inhibitor慢病毒的PANC-1组（1.002±0.160）与阴性病毒感染的PANC-1组（0.956±0.250）间差异无显著性，但慢病毒感染的PANC-1组的相对表达量（0.317±0.320）显著下降（P<0.05）。结论：miR-103a-3p在侵袭能力较强的胰腺癌细胞株中表达增高，miR-103a-3p-inhibitor能稳定转染PANC-1细胞系并抑制其miR-103a-3p表达。