lncRNA GAS6-AS1靶向调控IL-11对子宫内膜基质细胞向成纤维细胞分化的影响  

Effect of lncRNA GAS6-AS1 on differentiation of endometrial stromal cells into fibroblasts by regulating IL-11

作　　者：巫剑红[1] 代荫梅[1] 田玉翠[1] 蒋子雯 王静璇[1] 张宇迪[1] WU Jianhong;DAI Yinmei;TIAN Yucui;JIANG Ziwen;WANG Jingxuan;ZHANG Yudi(Department of Gynecology,Beijing Obstetrics and Gynecology Hospital,Capital Medical University,Beijing 100026,China)

机构地区：[1]首都医科大学附属北京妇产医院北京妇幼保健院妇科,北京100026

出　　处：《山东医药》2023年第13期10-14,共5页Shandong Medical Journal

基　　金：北京市研究型病房示范建设项目(BCRW202109);首都医科大学附属北京妇产医院北京妇幼保健院“优青人才”计划专项经费资助项目(YQRC201804)。

摘　　要：目的探讨长链非编码RNA(lncRNA)生长停滞特异基因转录的反义RNA(GAS6-AS1)靶向调控白细胞介素11(IL-11)对子宫内膜基质细胞(ESCs)向成纤维细胞分化的影响。方法体外传代培养ESCs,取传3~6代、对数生长期、生长状态良好的ESCs,随机分为TGF-β_(1)未干预组和TGF-β_(1)干预组。TGF-β_(1)未干预组不予TGF-β_(1)干预,TGF-β_(1)干预组用无血清培养基饥饿培养后予TGF-β_(1)干预,收集细胞,采用RT-qPCR法检测lncRNA GAS6-AS1、IL-11及纤维化标志物α平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白α1(COL1A1)mRNA表达。另取传3~6代、对数生长期、生长状态良好的ESCs,分别转染三条si-GAS6-AS1序列以及si-Control序列,采用RT-qPCR法验证转染效率。将转染si-GAS6-AS1的ESCs用无血清培养基饥饿培养后予TGF-β_(1)干预,收集细胞,采用Western blotting法检测IL-11、α-SMA、COL1A1蛋白表达,采用CCK-8法检测细胞增殖活性,采用Annexin V-FITC/PI双染法检测细胞凋亡率。结果与TGF-β_(1)未干预组比较,TGF-β_(1)干预组lncRNA GAS6-AS1、IL-11、α-SMA、COL1A1 mRNA相对表达量均显著升高(P均<0.01)。转染si-GAS6-AS1-1、si-GAS6-AS1-2、si-GAS6-AS1-3及si-Control序列的ESCs GAS6-AS1 mRNA相对表达量分别为0.22±0.04、0.27±0.06、0.81±0.07、1.00±0.00。与转染si-Control序列的ESCs比较,转染si-GAS6-AS1-1、si-GAS6-AS1-2序列的ESCs GAS6-AS1 mRNA相对表达量显著降低(P均<0.01),而转染si-GAS6-AS1-3序列的ESCs GAS6-AS1 mRNA相对表达量降低不明显(P>0.05)。因此,选择si-GAS6-AS1-1、si-GAS6-AS1-2序列进行后续实验。与si-Control+TGF-β_(1)组比较,si-GAS6-AS1-1+TGF-β_(1)组和si-GAS6-AS1-2+TGF-β_(1)组IL-11、α-SMA、COL1A1蛋白相对表达量及细胞增殖活性均显著降低,细胞凋亡率均显著升高(P均<0.01)。结论lncRNA GAS6-AS1通过靶向调控IL-11表达抑制ESCs向成纤维细胞分化。Objective To investigate the effect of long non-coding RNA(lncRNA)growth arrest-specific gene tran⁃scription antisense RNA(GAS6-AS1)on the differentiation of endometrial stromal cells(ESCs)into fibroblasts by regulat⁃ing interleukin-11(IL-11).Methods ESCs cultured in vitro in the 3rd-6th passage,which were in logarithmic growth phase and had good growth status,were randomly divided into TGF-β_(1) non-intervention group and TGF-β_(1) intervention group.ESCs in the TGF-β_(1) non-intervention group were not treated with TGF-β_(1),while ESCs in the TGF-β_(1) intervention group were treated with TGF-β_(1) after starvation culture in serum-free medium,and then cells were collected.The mRNA expression levels of lncRNA GAS6-AS1,IL-11 and fibrosis markersα-smooth muscle actin(α-SMA)and typeⅠcollagenα1(COL1A1)were detected by RT-qPCR.ESCs in the 3rd-6th passage,which were in logarithmic growth phase,and had good growth status,were transfected with three si-GAS6-AS1 and si-Control sequences,and RT-qPCR was used to verify the transfection efficiency.ESCs transfected with si-GAS6-AS1 were starved in serum-free medium and then subjected to TGF-β_(1) intervention.Cells were collected,and the protein expression levels of IL-11,α-SMA,and COL1A1 were detected by Western blotting.The proliferative activity of cells was detected by CCK-8 and the apoptosis rate was detected by Annexin V-FITC/PI double staining.Results The mRNA relative expression levels of lncRNA GAS6-AS1,IL-11,α-SMA and COL1A1 in the TGF-β_(1) intervention group were significantly higher than those in the TGF-β_(1) non-intervention group(all P<0.01).The relative mRNA expression levels of lncRNA GAS6-AS1 in ESCs transfected with si-GAS6-AS1-1,si-GAS6-AS1-2,si-GAS6-AS1-3 and si-Control were 0.22±0.04,0.27±0.06,0.81±0.07,and 1.00±0.00,respectively.Compared with ESCs transfected with si-Control,the relative mRNA expression levels of GAS6-AS1 in ESCs transfected with si-GAS6-AS1-1 and si-GAS6-AS1-2 decreased significantly(both P<0.01).However,the

关 键 词：宫腔粘连 长链非编码RNA生长停滞特异基因转录的反义RNA 白细胞介素11 子宫内膜基质细胞 成纤维细胞 

分 类 号：R711.74[医药卫生—妇产科学]