摘要
目的:探究长链非编码RNA(long-nocoding RNA,lncRNA)HOXA11-AS在骨肉瘤中的表达及其对骨肉瘤细胞增殖、迁移和侵袭的影响及其作用机制。方法:采用2012年1月至2015年12月浙江省人民医院骨外科13例骨肉瘤组织及癌旁正常组织,以及骨肉瘤细胞系MG63、U20S和人成骨细胞株h FOB1.19,用实时荧光定量PCR检测骨肉瘤组织和骨肉瘤细胞系MG63和U20S中lncRNA HOXA11-AS的表达水平。用慢病毒载体构建稳定过表达lncRNA HOXA11-AS的骨肉瘤MG63及U20S细胞,用CCK-8和克隆形成实验、Transwell小室法分别检测过表达lncRNA HOXA11-AS对骨肉瘤细胞增殖、迁移和侵袭的影响。建立过表达lncRNA HOXA11-AS MG63骨肉瘤细胞裸鼠移植瘤模型,以空载体慢病毒Lv-NC转染的细胞作为对照,观察过表达lncRNA HOXA11-AS对裸鼠移植瘤生长的影响。结果:lncRNA在骨肉瘤组织和MG63及U20S细胞中表达水平显著下调(P<0.01)。与h FOB1.19细胞比,过表达lncRNA HOXA11-AS细胞后,(1)显著抑制骨肉瘤MG63及U20S细胞的增殖[(4.03±0.98)vs(6.96±0.54),(4.68±0.77)vs(8.87±1.23),均P<0.01]、迁移细胞数[(83.00±6.03)vs(168±12.54),(96.00±8.77)vs(184.00±14.63)个,均P<0.01]和侵袭细胞数[(35.00±3.48)vs(97.00±8.32),(38.00±1.73)vs(87.00±6.37)个,均P<0.01];(2)显著抑制骨肉瘤裸鼠皮下移植瘤的生长(P<0.01)。结论:lncRNA HOXA11-AS在骨肉瘤细胞中低表达,且lncRNA HOXA11-AS过表达对骨肉瘤的发生发展具有抑制作用,可以作为骨肉瘤治疗潜在的分子靶点。
Objective: To investigate the expression of lncRNA HOXA11-AS in osteosarcoma,and to explore its effect on proliferation,migration and invasion of osteosarcoma cells and the possible mechanism. Methods: Thirteen cases of osteosarcoma tissues and corresponding adjacent normal tissues were collected from the Department of Bone Surgery,the People's Hospital of Zhejiang Province during January 2012 to December 2015. Osteosarcoma cell MG63,U205 lines and human osteoblast cell h FOB1. 19 line were used. Real-time fluorescent quantitative PCR was performed to detect the lncRNA HOXA11-AS expression in osteosarcoma specimens and osteosarcoma MG63 and U20 S cell lines. Lentiviral vectors that stably over-expressing lncRNA HOXA11-AS were constructed and transfected into MG63 and U20 S cell lines; CCK8 assay and colony-formation assay were performed to investigate the effect of lncRNA HOXA11-AS on osteosarcoma cell proliferation; Transwell assay was performed to investigate the effect of lncRNA HOXA11-AS on migration and invasion of osteosarcoma cells. Nude mice model of MG63 osteosarcoma xenograft,which over-expressing lncRNA HOXA11-AS,was established to observe the effect of lncRNA HOXA11-AS on xenograft growth in mice,by comparing with Lv-NC transfected cell xenografts. Results: lncRNA HOXA11-AS was markedly down-regulated in tumor tissues and osteosarcoma cell lines( P < 0. 01). Over-expression of lncRNA HOXA11-AS significantly suppressed( 1) the proliferation,migration and invasion of U20 S and MG63 cells( proliferation: [4. 03 ± 0. 98] times vs [6. 96 ± 0. 54] times,[4. 68 ± 0. 77] times vs[8. 87 ± 1. 23]times,all P < 0. 01; migration: [83. 00 ± 6. 03]vs [168 ± 12. 54],[96 ± 8. 77]vs [184 ± 14. 63],all P < 0. 01; invasion: [35. 00 ± 3. 48]vs [97. 00 ± 8. 32],[38. 00 ± 1. 73]vs [87. 00 ± 6. 37],all P < 0. 01);( 2) the growth of xenografts in rats( P < 0. 01). Conclusion: lncRNA HOXA11-AS significantly down-regulated in osteosarcoma cells,and suppressed the progression of osteosarcoma. lncRNA HOXA11-AS can be served as a therapeutic target for osteosarcoma.
引文
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