miR-34a-5p介导上皮-间充质转化促进胰腺癌细胞株PANC-1的侵袭转移及化疗抵抗研究

miR-34a-5p mediates epithelial-mesenchymal transformation to promote invasion,metastasis and chemotherapy resistance of PANC-1 pancreatic cancer cell line

目的 研究miR-34a-5p在胰腺癌细胞PANC-1中的表达水平以及其对上皮-间充质转化(EMT)、细胞侵袭、转移能力和化疗抵抗的影响。方法 构建PANC-1/miR-34a-5p细胞或PANC-1/mock细胞稳转株,将细胞分为4组:PANC-1组、PANC-1/miR-34a-5p组、PANC-1/mock组、PANC-1/miR-34a-5p+吉西他滨组。4组细胞均培养24 h。以实时定量聚合酶链反应法检测PANC-1细胞转染前后miR-34a-5p表达,用蛋白质印迹法检测EMT相关蛋白的表达,用伤口愈合实验检测细胞迁移能力,用Transwell法检测细胞侵袭能力,用细胞计数试剂盒-8(CCK-8)检测细胞增殖情况。结果 PANC-1组、PANC-1/mock组、PANC-1/miR-34a-5p组和PANC-1/miR-34a-5p+吉西他滨组的miR-34a-5p表达水平分别为1.01±0.08、1.03±0.06、2.43±0.15和1.76±0.21;E-cadherin水平分别为0.61±0.12、0.63±0.08、0.24±0.15和0.48±0.05;Vimentin水平分别为0.56±0.04、0.58±0.09、1.08±0.19和0.82±0.11;相对细胞迁移水平分别为(41.48±4.94)、(42.97±6.45)、(81.42±6.85)和(63.54±7.63)distance·μm^(-1);侵袭细胞分别为(212.62±9.80)、(209.46±13.67)、(463.83±35.22)和(325.46±28.44)个;增殖能力分别为(13.28±5.24)%、(14.37±4.79)%、(46.15±11.83)%和(35.78±9.42)%。PANC-1/miR-34a-5p组的上述指标与PANC-1组和PANC-1/mock组相比,在统计学上差异均有统计学意义(均P<0.05)。结论 miR-34a-5p过表达促进胰腺癌细胞EMT转化、促进胰腺癌细胞的迁移、促进胰腺癌细胞的侵袭、促进胰腺癌细胞的化疗抵抗。

Objective To explore the expression level of miR-34a-5p in pancreatic cancer cell PANC-1 and its effect on epithelial mesenchymal transition(EMT),cell invasion,metastasis,and chemotherapy resistance.Methods Construct stable transfected PANG-1/miR-34a-5p cells or PANC-1/mock cells,and divide the cells into four groups:PANC-1 group,PANC-1/miR-34a-5p group,PANC-1/mock group and PANC-1/miR-34a-5p+gemcitabine group.All four groups of cells were cultured for 24 h.Real-time quantitative polymerase chain reaction was used to detect the expression of miR-34a-5p in PANC-1 cells before and after transfection;Western blotting was used to detect the expression of EMT-related proteins;wound healing experiment was used to detect cell migration ability;the Transwell method was used to detect cell invasion ability;cell counting kit-8(GCK-8)test was used to detect cell proliferation.Results The expression levels of miR-34a-5p in PANC-1 group,PANC-1/mock group,PANC-1/miR-34a-5p group and PANC-1/miR-34a-5p+gemcitabine group were 1.01±0.08,1.03±0.06,2.43±0.15 and 1.76±0.21;the levels of E-cadherin were 0.61±0.12,0.63±0.08,0.24±0.15 and 0.48±0.05;the levels of Vimentin were0.56±0.34,0.58±0.29,1.08±0.19 and 0.82±0.11;the relative cell migration levels were(41.48±4.94),(42.97±6.45),(81.42±6.85)and(63.54±7.63)distance·μm^(-1);the number of invasive cells were212.62±9.80,209.46±13.67,463.83±35.22 and 325.46±28.44;the survival rates were(13.28±5.24)%,(14.37±4.79)%,(46.15±11.83)%and(35.78±9.42)%.The above indicators in the PANC-1/miR-34a-5p group showed statistically significant differences compared to the PANC-1 and PANC-1/mock groups(all P<0.05).Conclusion Overexpression of miR-34a-5p promotes EMT of pancreatic cancer cells,migration of pancreatic cancer cells,invasion of pancreatic cancer cells,and chemotherapy resistance of pancreatic cancer cells.