下调糖蛋白130基因表达对HaCaT细胞体外增殖及IL-6水平的影响  

Effects of Down Regulated Glycoprotein 130 Gene Expression on Proliferation and Expression of Interleukin 6 in HaCaT Cells

作　　者：孙海严 赵桂香[1] 冯世军[2] 张胜逆[1] 楚瑞琦[1] 

机构地区：[1]河北大学附属医院皮肤科,河北保定071000 [2]沧州市中心医院皮肤科,河北沧州061000

出　　处：《中国皮肤性病学杂志》2017年第10期1068-1072,共5页The Chinese Journal of Dermatovenereology

摘　　要：目的探讨下调糖蛋白(glycoprotein,gp)130基因表达对人永生化角质形成细胞株(human immortal keratinocyte colony HaCaT cells,HaCaT)细胞体外增殖及IL-6水平的影响。方法设计并合成三条gp130的siRNA,转染HaCaT细胞后,从mRNA表达、蛋白质水平上检测gp130,筛选出最适siRNA序列及浓度用于后续实验。转染HaCaT细胞24h,48h,MTT法检测细胞增殖情况;流式细胞仪检测细胞增殖周期;荧光定量PCR法检测mRNA表达;Western blotting印迹法检测蛋白水平;酶联免疫吸附法(ELISA)检测细胞上清液中IL-6水平。结果 siRNA-1与siRNA-NC、空白对照组相比,HaCaT细胞增殖率及增殖周期在24h时差异均无统计学意义(均P>0.05),48h时差异均有统计学意义(均P<0.05);IL-6mRNA表达、蛋白水平在转染HaCaT细胞24h时差异均无统计学意义(均P>0.05),48h时差异均有统计学意义(均P<0.05)。结论下调gp130基因可抑制角质形成细胞增殖,影响角质形成细胞增殖周期,并可抑制IL-6的表达和分泌。Objective To study the effect of down regulated glycoprotein 130 gene expression on the proliferation of HaCaT cells in vitro and the level IL-6 in vitro.Methods Three small interfering RNA of gp130 were designed and synthesized.After transfection into HaCaT cells,the expression of gp130 was detected by mRNA and protein level,the optimal siRNA sequence and concentration were selected for subsequent experiments.After 24 hours and 48 hours transfected,methylthiazoltetrazolium（MTT）assay was conducted to estimate the proliferation of HaCaT cells,flowcytometry used to detect cell proliferation; Quantitative fluorescent polymerase chain reaction（qPCR）to detect the messenger ribonucleic acid（mRNA）expression; Western blotting to detect protein expression; Enzyme linked immunosorbent assay（ELISA）to measure the level of IL-6 protein in the culture supernatant of HaCaT cells.Results The proliferation rate and proliferation cycle of HaCaT cells were not statistically significant at 24h,but the difference was statistically significant at 48h between siRNA-1 and siRNA-NC and blank control group（all P〈0.05）.The mRNA expression and protein secretion of IL-6 were not statistically significant at 24h,but the difference was statistically significant at 48h after transfection of HaCaT cells（all P〈0.05）.Conclusion Down regulation of gp130 gene expression can inhibit the proliferation and affect the proliferation cycle of keratinocytes,and inhibit the expression and secretion of IL-6.

关 键 词：角质形成细胞 白细胞介素6 糖蛋白130 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]