长链非编码RNA HIF2PUT影响骨肉瘤细胞迁移和侵袭过程

Effects of long-chain non-coding HIF2PUT on migration and invasion of osteosarcoma cells

目的观察长链非编码RNA(lncRNA)HIF2PUT对骨肉瘤细胞的增殖、迁移和侵袭过程的影响。方法采用实时荧光定量聚合酶链反应(FQ-PCR)检测2015年7月至2019年7月在郑州大学第一附属医院病理科确诊的47例(其中男28例,女19例,年龄最大者45岁,最小者8岁,平均年龄20.24岁)骨肉瘤组织及2种骨肉瘤细胞株(MG63和HOS)中lncRNA HIF2PUT的表达。通过慢病毒筛选稳转株后,根据细胞转染分别设置实验组(过表达lncRNA HIF2PUT组)、空白转染组、空白对照组。噻唑蓝(MTT)实验、细胞迁移实验(Transwell)检测lncRNA HIF2PUT对骨肉瘤细胞的增殖、迁移和侵袭过程的影响,应用SPSS 21.0统计软件分析。结果lncRNA HIF2PUT在47例骨肉瘤组织中的表达量(39.56±0.48)显著低于其在正常组织中的表达量(79.56±1.16,t=3.516,P<0.05),差异有统计学意义,且lncRNA HIF2PUT在骨肉瘤细胞株MG63中表达水平高于在HOS细胞株中的表达。细胞功能实验表明,lncRNA HIF2PUT对骨肉瘤细胞的增殖、迁移和侵袭过程产生显著影响。MG63和HOS细胞的增殖明显受到抑制,其增殖指数均低于空白转染组、空白对照组(t=4.785,P<0.05),差异有统计学意义。MG63组迁移实验中细胞穿透数目分别为(154.36±13.85)、(196.32±13.52)、(258.63±12.35)个(t=3.968,P<0.01),差异有统计学意义;HOS组迁移实验中细胞穿透数目分别为(131.24±10.58)、(247.36±17.52)、(224.52±14.52)个(t=3.052,P<0.01),差异有统计学意义。MG63组侵袭实验中细胞穿透数目分别为(73.50±13.89)、(98.63±12.57)、(112.30±15.29)个(t=23.178,P<0.01),差异有统计学意义;HOS组侵袭实验中细胞穿透数目分别为(70.74±13.49)、(98.40±16.46)、(89.74±12.47)个(t=14.275,P<0.01),差异有统计学意义。MG63和HOS细胞实验中迁移和侵袭能力均低于空白转染组、空白对照组。结论lncRNA HIF2PUT在骨肉瘤组织及细胞株中明显低表达,lncRNA HIF2PUT的表达明显抑制了骨肉细胞的增殖、迁移和侵袭过程。

Objective To observe the effects of long-chain non-coding RNA(lncRNA)HIF2PUT on the proliferation,migration and invasion of osteosarcoma cells.Methods Real-time fluorescent quantitative polymerase chain reaction(FQ-PCR)was used to detect the expression of lncRNA HIF2PUT in 47 cases of osteosarcoma tissues(28 males and 19 females,the oldest was 45 years old,the youngest was 8 years old with an average of 20.24 years old)and 2 osteosarcoma cell lines(MG63 and HOS).After screening for stable transfectants by lentivirus,the methyl thiazol tetrazolium(MTT)assay and Transwell assay were applied to detect the effects of lncRNA HIF2PUT on proliferation,migration and invasion of osteosarcoma cells.According to the cell transfection,the experimental group(overexpressed lncRNA HIF2PUT group),the blank transfection group and the blank control group were respectively set.Data were expressed as mean±standard deviations(SD),and SPSS 21.0 software was used for statistical analysis.Results The expression of lncRNA HIF2PUT in 47 cases of osteosarcoma(39.56±0.48)was significantly lower than that in normal tissues(79.56±1.16,t=3.516,P<0.05).The expression level of lncRNA HIF2PUT in MG63 cells was significantly higher than that in HOS cells.Cell function experiments showed that lncRNA HIF2PUT had a significant effect on the proliferation,migration and invasion of osteosarcoma cells.Proliferation of MG63 and HOS cells was significantly inhibited by lncRNA HIF2PUT,and the proliferation index was significantly lower in lncRNA HIF2PUT group than in blank transfection group and blank control group(t=4.785,P<0.05).For the migration experiment,the migrating number of MG63 cells was(154.36±13.85),(196.32±13.52)and(258.63±12.35),respectively(t=3.968,P<0.01),and that of HOS cells was(131.24±10.58),(247.36±17.52)and(224.52±14.52)respectively(t=3.052,P<0.01).For the invasion experiment,the penetrating number of MG63 cells was(73.50±13.89),(98.63±12.57)and(112.30±15.29),respectively(t=23.178,P<0.01),and that of HOS cells was(70.74±13.49)(98.40±16.46)and 89.74±12.47,respectively(t=14.275,P<0.01).The migration and invasion ability of MG63 and HOS cells in lncRNA HIF2PUT group was lower than that in blank transfection group and blank control group.Conclusion The expression of lncRNA HIF2PUT was significantly low in osteosarcoma tissues and MG63 and HOS cell lines.The expression of lncRNA HIF2PUT significantly inhibited the proliferation,migration and invasion of osteosarcoma cells.