CAJ | 학술논문

目的探讨含有WW结构域的氧化还原酶的基因(WWOX)的表达对人骨肉瘤细胞株MG-63的生长抑制作用及机制.方法构建WWOX重组真核表达质粒,转染入骨肉瘤细胞株MG-63,流式细胞仪膜联蛋白V/碘化丙锭(Annexin V/PI)双染色法检测MG-63的凋亡,逆转录-聚合酶链反应(RT-PCR)检测MG-63中B淋巴细胞/白血病-2(bcl-2)与WWOX基因的扩增,Western blot 法检测MG-63中bcl-2和WWOX蛋白的表达.结果建立稳定表达WWOX的骨肉瘤细胞模型,流式细胞仪Annexin V/PI双染法显示转染WWOX组骨肉瘤细胞较转染WWOX组和对照组凋亡明显增加[(8.21±0.03)％比(1.13±0.03)％比(1.12±0.02)％,P ＜0.05],转染WWOX组的WWOX mRNA表达明显增加(1.612±0.072),WWOX蛋白表达明显升高(1.680 ±0.072),与其他组比较差异有统计学意义(P＜0.05),转染WWOX组的bcl-2 mRNA表达明显降低(0.194±0.088),bcl-2蛋白表达明显降低(0.236 ±0.053),与其他组比较差异有统计学意义(P＜0.05).结论转染WOX的骨肉瘤细胞MG-63表达增加,bcl-2表达降低,骨肉瘤细胞的凋亡增加,可能与bcl-2的表达下调有关.
목적 탐토함유WW결구역적양화환원매적기인(WWOX)적표체대인골육류세포주MG-63적생장억제작용급궤제.방법 구건WWOX중조진핵표체질립,전염입골육류세포주MG-63,류식세포의막련단백V/전화병정(Annexin V/PI)쌍염색법검측MG-63적조망,역전록-취합매련반응(RT-PCR)검측MG-63중B림파세포/백혈병-2(bcl-2)여WWOX기인적확증,Western blot 법검측MG-63중bcl-2화WWOX단백적표체.결과 건립은정표체WWOX적골육류세포모형,류식세포의Annexin V/PI쌍염법현시전염WWOX조골육류세포교전염WWOX조화대조조조망명현증가[(8.21±0.03)％비(1.13±0.03)％비(1.12±0.02)％,P ＜0.05],전염WWOX조적WWOX mRNA표체명현증가(1.612±0.072),WWOX단백표체명현승고(1.680 ±0.072),여기타조비교차이유통계학의의(P＜0.05),전염WWOX조적bcl-2 mRNA표체명현강저(0.194±0.088),bcl-2단백표체명현강저(0.236 ±0.053),여기타조비교차이유통계학의의(P＜0.05).결론 전염WOX적골육류세포MG-63표체증가,bcl-2표체강저,골육류세포적조망증가,가능여bcl-2적표체하조유관.
Objective To investigate the effect of the tumor suppressor gene WW domain containing oxidoreductase (WWOX) expression on the growth of human osteosarcoma cell line (MG-63) and the mechanism.Methods The WWOX recombinant eukaryotic expression plasmid was constructed,and transfected into MG-63 cells.Flow cytometry was used to examine the apoptosis rate of MG-63 cells.Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to detect the expression of WWOX and B cell lymphoma/lewkmia-2 (bcl-2).Results Osteosarcoma cells with stable expression of WWOX were build.Flow cytometry revealed that the apoptosis rate of osteosarcoma cells in WWOX-transfected group was significantly increased as compared with blank plasmid group and control group [(8.21±0.03)％ vs.(1.13±0.03)％ vs.(1.12±0.02)％,P＜0.05].The expression of WWOX mRNA (1.612 ± 0.072) and protein (1.680 ± 0.072) was increased significantly in WWOX-transfected group as compared with the other groups (P ＜0.05).The expression of bcl-2 mRNA (0.194 ±0.088) and protein (0.236 ± 0.053) was decreased significantly in WWOX-transfected group as compared with the other groups (P ＜0.05).Conclusion WWOX can inhibit the proliferation of osteosarcoma cells and promote apoptosis of osteosarcoma cells,which may be associated with the decreased expression of bcl-2.