CAJ | 학술논문

目的研究Hic-5/ARA55对大肠癌细胞生长的影响及其机制.方法流式细胞仪检测已稳定转染Hic-5/ARA55的大肠癌细胞系(Lovo-Hic-5/ARA55组)的细胞周期,用未转染质粒(Lovo组)和转染空质粒的Lovo细胞(Lovo-Vector组)作为对照.Western blot方法检测各组细胞之间主要细胞周期蛋白的表达差异,Luciferase Assay进一步研究Hic-5/ARA55与细胞周期蛋白的作用机制.建立裸鼠皮下种植瘤模型,7周后切取瘤体并称重,应用免疫组织化学方法检测各组皮下种植瘤的Hic-5/ARA55及相关细胞周期蛋白的表达差异.结果 Lovo-Hic-5/ARA55组细胞由G0/G1期进入S期明显延迟,并且细胞周期蛋白P27表达升高,Luciferase Assay证明Hic-5/ARA55在转录水平上调P27的表达.Lovo-Hic-5/ARA55组皮下种植瘤质量[(0.33±0.23)g]明显低于Lovo组[(1.20±0.39)g]和Lovo-Vector组[(1.30±0.49)g],差异有统计学意义(P＜0.05).Lovo-Hic-5/ARA55组种植瘤Hic-5/ARA55和P27均高表达,而Lovo组和Lovo-Vector组均低表达或阴性表达,差异有统计学意义(P＜0.05),并且Hic-5/ARA55和P27的表达呈正相关(r=0.816,P＜0.05).结论 Hic-5/ARA55在体外和裸鼠体内均通过在转录水平上调细胞周期抑制蛋白P27的表达来抑制大肠癌细胞的生长.
목적 연구Hic-5/ARA55대대장암세포생장적영향급기궤제.방법 류식세포의검측이은정전염Hic-5/ARA55적대장암세포계(Lovo-Hic-5/ARA55조)적세포주기,용미전염질립(Lovo조)화전염공질립적Lovo세포(Lovo-Vector조)작위대조.Western blot방법검측각조세포지간주요세포주기단백적표체차이,Luciferase Assay진일보연구Hic-5/ARA55여세포주기단백적작용궤제.건립라서피하충식류모형,7주후절취류체병칭중,응용면역조직화학방법검측각조피하충식류적Hic-5/ARA55급상관세포주기단백적표체차이.결과 Lovo-Hic-5/ARA55조세포유G0/G1기진입S기명현연지,병차세포주기단백P27표체승고,Luciferase Assay증명Hic-5/ARA55재전록수평상조P27적표체.Lovo-Hic-5/ARA55조피하충식류질량[(0.33±0.23)g]명현저우Lovo조[(1.20±0.39)g]화Lovo-Vector조[(1.30±0.49)g],차이유통계학의의(P＜0.05).Lovo-Hic-5/ARA55조충식류Hic-5/ARA55화P27균고표체,이Lovo조화Lovo-Vector조균저표체혹음성표체,차이유통계학의의(P＜0.05),병차Hic-5/ARA55화P27적표체정정상관(r=0.816,P＜0.05).결론 Hic-5/ARA55재체외화라서체내균통과재전록수평상조세포주기억제단백P27적표체래억제대장암세포적생장.
Objective To investigate the effects of Hic-5/ARA55 on the growth of the human colorectal cancer cells(Lovo cells)and its mechanism.Method Flow cytometry(FCM)was used to study the cell cycle of Lovo cells(Lovo group),Lovo cells stably transfected with empty vector(Lovo-Vector group)and the Lovo cells stably transfected with vector containing Hic-5/ARA55(Lovo-Hic-5/ARA55group).Western blot assay was used to detect the principal cyclins in the three groups,and Luciferase assay was used to study the mechanism between Hic-5/ARA55 and the only target cyclin. The cells from the three groups were inoculated subcutaneously into 7 nude micef Babl/c nu/nu)respectively to observe the effects of Hic-5/ARA55 on the growth of the cells in vivo.Seven weeks later,the subcutaneous tumors were harvested and weighed.Then immunohistochemistrical assay was used to detect Hic-5/ARA55 and the target cyclin in the tumors.Results The cell cycle was obviously delayed from G0/G1 to S stage in Lovo-Hic-5/ARA55 cells.A significantly higher expression of P27 was found in Lovo-Hic-5/ARA55 cells than in the other two groups.The weight of the subcutaneous tumors of Lovo-Hic-5/ARA55 cells,Lovo cells and Lovo-Vector cells were(0.33±0.23)g,(1.20±0.39)g and(1.30 ±0.49)g,respectively;the tumors of Lovo-Hic-5/ARA55 cells was significantly lighter than those ofthe other two groups(P＜0.05).Hic-5/ARA55 and P27 were beth over-expressed in implanted tumors of Lovo-Hic-5/ARA55 cells,while were both expressed lower or not expressed in the other two groups.And the expressions of Hic-5/ARA55 and P27 were highly positive correlated(r=0.816,P＜0.05).Conclusion Hic-5/ARA55 could inhibit the growth of Lovo cells both in vitro and in vlvo bv up-regulating the transcription of P27.