摘要
目的观察骨囊肿与骨肉瘤组织中Y染色体上的性别决定区相关高迁移率族盒蛋白-5（SOX-5）的表达，探讨SOX-5对骨肉瘤细胞MG63增殖的影响。方法通过反转录-聚合酶链反应（RT—PCR）检测骨囊肿与骨肉瘤组织中的SOX-5表达，进而通过RNA干扰降低SOX-5在骨肉瘤细胞MG63中的表达，再采用细胞计数试剂盒（CCK-8）法分别检测干扰后0、24、48、72h的细胞增殖率并分别与对照组比较；流式细胞分析评估细胞周期分布，Westernblot检测干扰组和对照组细胞周期调控蛋白细胞周期蛋白依赖性激酶2（CDK2）和细胞分裂周期蛋白25A（CDC25A）的表达水平。结果与骨囊肿组织比较，SOX-5mRNA在骨肉瘤组织显著上升（P〈0．001）。RNA干扰后的骨肉瘤细胞的增殖明显降低（P〈0．01），SOX-5干扰组的G0／G1期细胞比例从（52．80±1．43）％增加至（66．71±0．62）％（P〈0．01），而S期细胞比例从（30．72±1．72）％下降至（17．27±3．15）％（P〈0．01）。SOX-5干扰组的CDK2和CDC25A表达均显著降低（P〈0．01），SOX-5的低表达降低了G1／S期的细胞周期过渡。结论SOX-5在骨肉瘤细胞中高表达，它能促进肿瘤细胞的增殖。
Objective To observe the expression of sex determination region related high mobility group box protein- 5 （SOX- 5 ） in bone cysts and osteosarcoma, and its effect on the proliferation of MG63 cells. Methods The expression of SOX -5 in bone cysts and MG63 cells was detected by reverse transcriptase- polymerase chain reaction （RT- PCR）. The expression of SOX-5 in osteosarcoma cells was reduced by RNA interference. The proliferation of the cells at 0, 24, 48, and 72 h after interference was detected by cell counting kit （ CCK - 8） assay and compared with the control group respectively. Cell cycle distribution was detected by flow cytometry. The expression level of cell division cyclin （CDC25A） and cyclin - dependent kinase （CDK2）, cell cycle regulatory proteins, was detected by Western blotting. Results As com- pared with bone cysts, SOX -5 mRNA expression was increased significantly in osteosareoma cells （P 〈0. 01 ）. The proliferation of osteosarcoma cells after RNA interference was significantly reduced （P 〈 0. 01 ）. SOX - 5 knockdown in MG63 cells increased the proportion of G0/G1 phase cells from （52. 80 ± 1.43）% to （66. 71 ± 0. 62）% （P 〈0. 01 ）, while decreased the proportion of S phase ceils from （30. 72 ± 1.72）% to （ 17.27 ± 3. 15）% （P 〈0. 01 ）. The expression of CDK2 and CDC25A significantly reduced in SOX- 5 interference group （P 〈0. 01 ）. Low expression of SOX -5 reduced G1/S phase of the cell cycle transition. Conclusion SOX - 5 is over - expressed in osteosarcoma cells and it can promote the proliferation of tumor cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第7期1744-1746,共3页
Chinese Journal of Experimental Surgery
