miR-181a通过靶向PTEN促进软骨肉瘤细胞的生长  

MiR-181a promotes the growth of chondrosarcoma cells by targeting PTEN

作　　者：何畔[1] 汪志军 曾旭凯 He Pan;Wang Zhijun;Zeng Xukai(Department of Orthopedics,Hunan Provincial People's Hospital(The First Affiliated Hospital of Hunan Normal University),Changsha 410005,China)

机构地区：[1]湖南省人民医院(湖南师范大学附属第一医院)骨科,长沙410005

出　　处：《中国医师杂志》2023年第4期541-545,共5页Journal of Chinese Physician

基　　金：湖南省自然科学基金(2020JJ4402)。

摘　　要：目的探讨miR-181a通过磷酸酶和张力蛋白同源物(PTEN)促进软骨肉瘤细胞生长的作用及其可能的调控机制。方法收集2022年1月至12月湖南省人民医院骨科患者手术切除的新鲜软骨肉瘤及软骨瘤组织各10例,采用实时荧光定量PCR(qRT-PCR)法分别检测miR-181a在软骨肉瘤及软骨瘤组织中的表达;体外培养软骨肉瘤细胞SW1353,分别转染miR-181a抑制剂(miR-181a抑制组)及其对照(对照miR-NC)到细胞中,通过CCK-8细胞计数实验及克隆形成实验分别检测miR-181a对SW1353细胞生长及增殖能力的影响;通过Target Scan数据库预测miR-181a与PTEN之间的结合位点,并采用双荧光素酶报告基因实验进行验证;分别转染miR-181a模拟物(miR-181a组)及其对照(miR-NC组)到软骨肉瘤细胞SW1353中,检测细胞中ATP含量、葡萄糖消耗量和乳酸生成量,分析miR-181a对SW1353细胞糖酵解的影响。结果软骨肉瘤组织中miR-181a的表达明显高于软骨瘤组织(P<0.05);miR-181a抑制组的细胞生长及克隆形成能力均明显低于对照组(均P<0.05);经Target Scan在线网站预测,miR-181a与PTEN之间可能存在结合位点,且双荧光素酶报告基因实验证实共转染野生型PTEN和miR-181a可显著降低荧光素酶活性(P<0.05);miR-181a组的ATP含量、葡萄糖消耗量和乳酸生成量均明显高于miR-NC组(均P<0.05)。结论miR-181a通过PTEN介导糖酵解促进软骨肉瘤细胞的生长。Objective To explore the effect of miR-181a on chondrosarcoma cell growth through phosphatase and tensin homolog(PTEN)and its possible regulatory mechanism.Methods From January to December 2022,10 fresh chondrosarcoma and 10 chondroma tissues from orthopedic patients of Hunan Provincial People's Hospital were collected,and the expression of miR-181a in chondrosarcoma and chondroma tissues was detected using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR);Chondrosarcoma cell SW1353 was cultured in vitro and transfected with miR-181a inhibitor(miR-181a inhibition group)and control(miR-NC,control group),respectively.The effects of miR-181a on the growth and proliferation of SW1353 cells were detected by cell counting kit(CCK-8)and clone formation test,respectively;The binding sites between miR-181a and PTEN were predicted through the Target Scan database,and verified using dual luciferase reporter gene experiments;The mimetic miR-181a(miR-181a group)and its control(miR-NC,control group)were transfected into chondrosarcoma cell SW1353,respectively.The adenosine triphosphate(ATP)content,glucose consumption,and lactic acid production in the cells were measured,and the effect of miR-181a on glycolysis of SW1353 cells was analyzed.Results The expression of miR-181a in chondrosarcoma tissues was significantly higher than that in chondroma tissues(P<0.05).The cell growth and clonogenesis ability of miR-181a inhibition group were significantly lower than those of control group(all P<0.05).It was predicted by Target Scan online website that there might be binding sites between miR-181a and PTEN,and co-transfection of wild-type PTEN and miR-181a could significantly reduce luciferase activity by double luciferase reporter assay(P<0.05).The ATP content,glucose consumption and lactic acid production of miR-181a group were significantly higher than those of miR-NC group(all P<0.05).Conclusions MiR-181a promotes the growth of chondrosarcoma cells through PTEN-mediated glycolysis.

关 键 词：骨肉瘤 miR-181a PTEN磷酸水解酶 糖酵解 

分 类 号：R738.1[医药卫生—肿瘤]