摘要
目的研究过表达miR-29a-3p对IL-22诱导的HaCaT细胞增殖的影响。方法将HaCaT细胞分为Cell组、IL-22组、IL-22+NC组和IL-22+miR-29a-3p组,荧光定量PCR检测miR-29a-3p的表达水平,CCK8检测细胞的活力,流式细胞仪检测细胞凋亡及周期。结果与0μg/L组相比,25μg/L组、50μg/L组和100μg/L组HaCaT细胞的增殖率在24 h、48 h和72 h均出现升高(F值分别为33.27、36.19、52.29,均P<0.0001)。与0μg/L组相比,miR-29a-3p在50μg/L组和100μg/L组HaCaT中的表达水平降低(F=129,P<0.0001),分别降低83%和80%。与IL-22+NC组相比,IL-22+miR-29a-3p组的增殖率在24 h、48 h和72 h均降低(P值分别为0.0021、0.0016、0.0231),细胞总凋亡率增加(6.67±1.06 vs 30.55±1.86,P=0.0001),G1期细胞比例增加(P=0.0001),S期细胞比例降低(P=0.0001)。结论IL-22可降低HaCaT中miR-29a-3p的表达量,过表达miR-29a-3p通过促进凋亡和引起细胞G1期阻滞抑制IL-22诱发的HaCaT细胞过度增生。
Objective To investigate the effect of miR-29a-3p overexpression on IL-22-induced proliferation of HaCaT cells.Methods HaCaT cells were divided into four groups,Cell group,IL-22 group,IL-22+NC group and IL-22+miR-29a-3p group.The expression level of miR-29a-3p was detected by fluorescence quantitative PCR.Cell viability was detected by CCK8.Apoptosis and cell cycle were detected by flow cytometry.Results Compared with the 0 g/L group,the proliferation rate of HaCaT cells in the 25μg/L group,50μg/L group and 100μg/L group was increased at 24 h,48 h and 72 h(F value was 33.27,36.19,52.29,respectively,all P<0.0001).Compared with the 0μg/L group,miR-29a-3p expression level in HaCaT in 50μg/L and 100μg/L groups was decreased(F=129,P<0.0001),with a decrease of 83%and 80%,respectively.Compared with the IL-22+NC group,proliferation rate of IL-22+miR-29a-3p group was decreased at 24 h,48 h and 72 h(P value was 0.0021,0.0016,0.0231,respectively),total apoptosis rate was increased(6.67±1.06 vs 30.55±1.86,P=0.0001),cell proportion in G1 phase was increased(P=0.0001),and the cell proportion in S phase was decreased(P=0.0001).Conclusion Il-22 can reduce miR-29a-3p expression level in HaCaT,and miR-29a-3p overexpression can inhibit the excessive proliferation induced by IL-22 in HaCaT cells by promoting apoptosis and inducing G1 phase arrest.
作者
李璟蓉
思远
赵瑞
吴江
方锐华
LI Jingrong;SI Yuan;ZHAO Rui;WU Jiang;FANG Ruihua(Department of Dermatology,Guangzhou First People's Hospital,School of Medicine,South China Universiey of Technology,Guangzhou 510180,China)
出处
《广州医药》
2020年第3期26-30,共5页
Guangzhou Medical Journal
基金
广东省医学科研基金项目(A2018556)
广州市卫生计生科技一般引导项目(20181A010005)
广东省基础与应用基础研究基金(2019A1515010250)。