miR-581对结直肠癌细胞增殖与侵袭的影响及机制

Effects of miR-581 on proliferation and invasion of colorectal cancer cells

目的探究miR-581对结直肠癌细胞增殖与侵袭的影响及机制。方法取处于对数生长期的结肠癌细胞株SW620、LOVO、HT29和正常结直肠细胞株FHC,将细胞分为实验组和对照组,实验组加入miR-581 mimics,对照组加入vector;采用qRT-PCR法检测不同结直肠癌细胞株SW620、LOVO、HT29和正常结直肠细胞株FHC中miR-581的表达;CCK-8、Transwell侵袭实验分别检测miR-581 mimics组和对照组结直肠癌细胞SW620增殖和侵袭能力的变化;荧光素酶报告基因验证miR-581与过氧化物氧化还原酶蛋白1(PRDX1)的相互作用关系;Western blotting法检测miR-581对SW620细胞中PRDX1表达的影响。将SW620细胞分为两组,miR-581 mimics组转染miR-581mimics,miR-581+PRDX组将PRDX1质粒与miR-581 mimics共转染,CCK-8、Transwell侵袭实验检测PRDX1干预后miR-581对SW620增殖和侵袭能力的影响。结果与结直肠癌细胞株LOVO、HT29相比,miR-581在高侵袭性细胞株SW620中表达最低;SW620细胞转染miR-581 mimics后,实验组细胞增殖能力和侵袭能力低于对照组(P均<0.05)。miR-581能与PRDX1 3'UTR特异性结合,抑制PRDX1蛋白表达;miR-581+PRDX1组细胞的增殖和侵袭能力低于miR-581 mimics组(P均<0.05)。结论 miR-581可抑制结直肠癌细胞的增殖和侵袭,其机制可能与靶向调控PRDX1有关。

Objective To investigate the effects and mechanism of miR-581 on proliferation and invasion of colorectal cancer cells. Methods The colon cancer cell lines SW620,LOVO,HT29,and the normal colorectal cell line FHC in the logarithmic growth phase were selected and then we divided the cells into the experimental group and control group. We added miR-581 mimics to the cells of the experimental group,and added vector to the cells of the control group. qRT-PCR was used to detect the expression of miR-581 in different colorectal cancer cell lines SW620,LOVO,HT29 and the normal colorectal cell line FHC. CCK-8 and Transwell invasion assays were used to detect the proliferation and invasion of colorectal cancer cell line SW620 in the miR-581 mimics group and the control group,respectively. The luciferase reporter gene verified the interaction between miR-581 and peroxiredoxin 1（ PRDX1）. The effect of miR-581 on the expression of PRDX1 in SW620 cells was detected by Western blotting. SW620 cells were divided into two groups： the miR-581 mimics group which was transfected with miR-581 mimics,and the miR-581 ＋ PRDX1 group which was co-transfected with miR-581 mimics and PRDX1 plasmid. CCK-8 and Transwell invasion assays were used to detect the effect of miR-581 on the proliferation and invasion of SW620 cells after PRDX1 intervention. Results Compared with the colorectal cancer cell lines LOVO and HT29,miR-581 expressed the lowest in the highly invasive cell line SW620. After miR-581 mimics was transfected into SW620 cells,the cell proliferation and invasion abilities of the experimental group were significantly lower than that of the control group. miR-581 specifically bound to PRDX1 3＇UTR and inhibited PRDX1 protein expression. The miR-581 mimics ＋ PRDX1 group had lower proliferation and invasion than those of the miR-581 mimics group（ both P〈0. 05）. Conclusion miR-581 can inhibit the proliferation and invasion of colorectal cancer cells,and its mechanism may be related to the targeted regulation of PRDX1.