miR―103a―3p在胰腺癌细胞中表达及意义.docVIP

miR―103a―3p在胰腺癌细胞中表达及意义 　　【摘要】 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（P0.05）。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（P0.05）。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。 　　【关键词】 miR-103a-3p； 胰腺癌； 慢病毒 　　【Abstract】 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 signifi