LncRNA MALAT1调节miR-383-5p/SOCS3轴对人牙周膜干细胞的影响  

Impacts of LncRNA MALAT1 on human periodontal ligament stem cells by regulating miR-383-5p/SOCS3 axis

作　　者：顾婷立 刘亚华 钱靓[1] 章茜 GU Tingli;LIU Yahua;QIAN Liang;ZHANG Qian(Department of Oral and Maxillofacial Surgery,Nanjing Stomatological Hospital,Medical School of Nanjing University,Nanjing 210008,China)

机构地区：[1]南京大学医学院附属口腔医院颌面外科,江苏南京210008

出　　处：《口腔医学》2022年第11期966-973,共8页Stomatology

基　　金：国家自然科学基金(82103516)。

摘　　要：目的探究长链非编码RNA(long non-coding RNA,LncRNA)肺腺癌转移相关转录本1(metastasis-associated lung adenocarcinoma transcript 1,MALAT1)调控微小RNA-383-5p(microRNA-383-5p,miR-383-5p)/细胞因子信号转导负调控因子3(suppressor of cytokine signaling 3,SOCS3)轴对脂多糖(lipopolysaccharide,LPS)刺激的人牙周膜干细胞(human periodontal ligament stem cells,hPDLSCs)增殖、凋亡及成骨分化的影响。方法LPS处理hPDLSCs后用靶向MALAT1的小干扰RNA(small interfering RNA-MALAT1,si-MALAT1)或miR-383-5p抑制物(miR-383-5p inhibitor,in-miR-383-5p)或SOCS3过表达质粒(SOCS3)转染,ELISA检测炎症因子水平;RT-qPCR和Western blot检测转染效率;CCK-8和TUNEL检测细胞增殖、凋亡;使用碱性磷酸酶(alkaline phosphatase,ALP)染色和茜素红S(alizarin red S,ARS)染色分析成骨分化;Western blot对增殖、凋亡以及成骨分化相关蛋白进行检测。生物信息学与双荧光素酶实验分析miR-383-5p与MALAT1/SOCS3之间的关系。结果LPS可诱导hPDLSCs细胞炎症和凋亡,降低了细胞增殖能力和成骨分化程度;同时上调MALAT1和SOCS3表达,下调miR-383-5p表达(P<0.05)。敲低MALAT1可明显降低LPS诱导的hPDLSCs细胞炎症反应、凋亡能力,细胞增殖能力和成骨分化程度增加;同时还可上调miR-383-5p表达,下调SOCS3表达(P<0.05)。此外,回复实验表明,抑制miR-383-5p表达或上调SOCS3表达可部分逆转MALAT1敲低后对LPS诱导的hPDLSCs细胞的影响(P<0.05)。进一步分析验证,MALAT1海绵化miR-383-5p靶向调控SOCS3表达(P<0.05)。结论敲低MALAT1可通过调节miR-383-5p/SOCS3轴改善炎症状态下hPDLSCs的增殖、凋亡和成骨分化。Objective To investigate the impacts of long non-coding RNA(LncRNA)metastasis-associated lung adenocarcinoma transcript 1(MALAT1)on the proliferation,apoptosis and osteogenic differentiation of lipopolysaccharide(LPS)-stimulated human periodontal ligament stem cells(hPDLSCs)by regulating the microRNA-383-5p(miR-383-5p)/suppressor of cytokine signaling 3(SOCS3)axis.Methods hPDLSCs were treated with LPS and transfected with small interfering RNA targeting MALAT1(si-MALAT1)or miR-383-5p inhibitor(in-miR-383-5p)or SOCS3 overexpression plasmid(SOCS3).ELISA was implemented to detect inflammatory factor levels;RT-qPCR and Western blot were used to check transfection efficiency;CCK-8 and TUNEL were implemented to detect cell proliferation and apoptosis;alkaline phosphatase(ALP)staining and alizarin red S(ARS)staining were performed to analyze osteogenic differentiation;Western blot was implemented to measure the proliferation,apoptosis and osteogenic differentiationrelated proteins.Bioinformatics and dual-luciferase experiments were performed to analyze the relationship between miR-383-5p and MALAT1/SOCS3.Results LPS induced the inflammation and apoptosis of hPDLSCs,and down-regulated cell proliferation and osteogenic differentiation.In the meantime,the expressions of MALAT1 and SOCS3 were up-regulated,and the expression of miR-383-5p was down-regulated(P<0.05).Knockdown of MALAT1 could greatly reduce LPS-induced inflammatory response and apoptosis of hPDLSCs,and increase cell proliferation and osteogenic differentiation.Meanwhile,it also up-regulated the expression of miR-383-5p and down-regulated the expression of SOCS3(P<0.05).In addition,reverse experiments showed that inhibiting miR-383-5p expression or up-regulating SOCS3 expression could partially reverse the effects of MALAT1 knockdown on LPS-induced hPDLSCs(P<0.05).Further analysis confirmed that MALAT1 targeted the regulation of SOCS3 expression by sponging miR-383-5p(P<0.05).Conclusion Knockdown of MALAT1 can modulate the miR-383-5p/SOCS3 axis to improve the p

关 键 词：长链非编码RNA肺腺癌转移相关转录本1 微小RNA-383-5p 细胞因子信号转导负调控因子3 人牙周膜干细胞 

分 类 号：R781.4[医药卫生—口腔医学]