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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 long-nocoding RNA (HOXA11-AS) in osteosarcoma and its effect on the proliferation, migration and invasion of osteosarcoma cells and its mechanism. METHODS: Thirteen osteosarcoma tissues and adjacent normal tissues from osteosarcoma of osteosarcoma cell line MG63, U20S and human osteoblast cell line hFOB1.19 were obtained from January 2012 to December 2015 in Zhejiang Provincial People’s Hospital. The expression of lncRNA HOXA11-AS in osteosarcoma and osteosarcoma cell lines MG63 and U20S was detected by real-time fluorescence quantitative PCR. The osteosarcoma MG63 and U20S cells stably overexpressing lncRNA HOXA11-AS were constructed by lentiviral vector. The proliferation, migration and invasion of osteosarcoma cells were detected by Transwell chamber assay using CCK-8 and clone formation assay respectively influences. To investigate the effect of lncRNA HOXA11-AS on the growth of transplanted xenografts in nude mice, we constructed a transplanted nude mouse model of overexpressing lncRNA HOXA11-AS MG63 osteosarcoma cells. Results: The expression of lncRNA in osteosarcoma and MG63 and U20S cells was significantly down-regulated (P <0.01). Compared with h FOB1.19 cells, (1) significantly inhibited the proliferation of osteosarcoma MG63 and U20S cells (4.03 ± 0.98 vs 6.96 ± 0.54 vs 4.68 ± 0.77 vs (P <0.01). The number of migrating cells [(83.00 ± 6.03) vs (168 ± 12.54), (96.00 ± 8.77) vs (184.00 ± 14.63) (35.00 ± 3.48 vs 97.00 ± 8.32 vs 38.00 ± 1.73 vs 87.00 ± 6.37, P <0.01 respectively). (2) The growth of osteosarcoma xenografts in nude mice was significantly inhibited (P <0.01) . CONCLUSION: The lncRNA HOXA11-AS is overexpressed in osteosarcoma cells and overexpression of lncRNA HOXA11-AS can inhibit the development of osteosarcoma and may be used as a potential molecular target for the treatment of osteosarcoma.