摘要
目的探究miR-96通过干扰裸露角质蛋白同源物2(naked cuticle homolog 2,NKD2)的表达对骨肉瘤MG63细胞株的增殖、凋亡的影响。方法回顾性分析我院2013年9月至2018年2月收集的骨肉瘤标本57例临床资料作为研究组,同时收集距肿瘤边界5 cm的癌旁组织作为对照组,其中男39例,女18例;年龄21~49岁,平均(28.16±6.73)岁。将骨肉瘤MG63细胞株分为miR-96抑制组与空白对照组,其中miR-96抑制组细胞给予miR-96抑制剂转染,空白对照组细胞不作任何处理。培养48h后,采用实时荧光定量聚合酶链式反应(real time quantitative polymerase chain reaction,RT-qPCR)法与Western blot法检测各组人骨肉瘤MG63细胞中miR-96与NKD2蛋白表达水平;采用四甲基偶氮唑盐(methyl thiazolyltetrazolium assay,MTT)法检测各组细胞增殖情况;采用流式细胞仪检测各组细胞凋亡情况与细胞周期分布情况。结果骨肉瘤组织中miR-96表达水平明显高于正常骨组织(P<0.05);骨肉瘤组织中NKD2蛋白表达水平明显低于正常骨组织(P<0.05);miR-96抑制组骨肉瘤细胞中miR-96表达水平明显低于空白对照组(P<0.05);miR-96抑制组骨肉瘤细胞中NKD2表达水平明显高于空白对照组(P<0.05);miR-96抑制组骨肉瘤MG63细胞株经miR-96抑制剂转染48h后细胞相对吸光度明显低于空白对照组细胞(P<0.05);miR-96抑制组骨肉瘤MG63细胞株细胞凋亡率明显大于空白对照组(P<0.05);miR-96抑制组与空白对照组骨肉瘤MG63细胞株细胞周期情况差异明显(P<0.05)。结论miR-96在骨肉瘤组织中呈高表达,NKD2在骨肉瘤组织中呈低表达;miR-96通过抑制NKD2表达而促进骨肉瘤MG63细胞株生长,并抑制骨肉瘤MG63细胞株早期凋亡。
Objective To investigate the effect of miR-96 on proliferation and invasion of osteosarcoma MG63 cell line by interfering with naked cuticle homolog 2(NKD2)expression.Methods The clinical data of 57 osteosarcoma specimens collected in our hospital from September 2013 to February 2018 were retrospectively analyzed as the study group,and the adjacent tissues 5cm away from the tumor boundary were collected as the control group,including 39 males and 18 females.The age ranged from 21 to 49,with an average age of(28.16±6.73).Osteosarcoma MG63 cells were divided into miR-96 inhibition group and blank control group.After 48 hours of culture,the expression levels of miR-96 and nkd2 in MG63 cells were detected by RT-qPCR and Western blot.MTT method was used to detect the cell proliferation.Cell apoptosis and cell cycle distribution were detected by flow cytometry.Results The expression level of miR-96 in osteosarcoma tissue was significantly higher than that in normal bone tissue(P<0.05).The expression level of NKD2 protein in osteosarcoma tissue was significantly lower than that in normal bone tissue(P<0.05).The expression level of miR-96 in osteosarcoma cells in the miR-96 inhibition group was significantly lower than that in the blank control group(P<0.05).The expression level of NKD2 in osteosarcoma cells in miR-96 inhibition group was significantly higher than that in the blank control group(P<0.05).The relative absorbance of osteosarcoma MG63 cell lines in the miR-96 inhibition group was significantly lower than that in the blank control group(P<0.05).The apoptosis rate of osteosarcoma MG63 cell line inmiR-96 inhibition group was significantly higher than that in the blank control group(P<0.05).The cell cycle of osteosarcoma MG63 cell line was significantly different between the miR-96 inhibition group and the blank control group(P<0.05).Conclusion miR-96 is highly expressed in osteosarcoma,and NKD2 is down-regulated in osteosarcoma.miR-96 promotes the growth and invasion of osteosarcoma MG63 cell line by inhibitin
作者
张建业
张卫华
Zhang Jianye;Zhang Weihua(1st Department of Orthopedics,Hanyang Hospital Affiliated to Wuhan University of Science and Technology,Hanyang Hospital,Wuhan 430000,China)
出处
《实用骨科杂志》
2021年第7期615-618,631,共5页
Journal of Practical Orthopaedics
基金
湖北省卫生健康委员会2019—2020年度面上项目和血防专项(WJ2019M038)。
