目的:研究RACK1高表达对人结肠癌细胞增殖能力的影响。方法:采用脂质体转染术分别建立R ACK1表达下调的S W620细胞系、R ACK1表达上调的S W480细胞系以及对照细胞系;采用CCK-8细胞增殖测定、软琼脂集落形成实验、Ed U掺入实验以及流式细胞术检测RACK1表达改变对S W620和S W480细胞增殖的影响;采用Wester n blot分析R ACK1表达改变对G1/S期限制点调控蛋白Cyclin D1和p27蛋白表达的影响。结果:下调RACK1的表达抑制SW620细胞的生长、软琼脂集落形成能力,降低Ed U标记的S期细胞数目,阻滞细胞周期于G1/S期;而上调RACK1的表达增强S W 4 8 0细胞的生长、软琼脂集落形成能力,增加Ed U标记的S期细胞数目,促进细胞周期G 1/S期进程。结论:RACK1高表达促进人结肠癌细胞的生长和增殖。
Objective: To investigate the effects of RACK1 overexpression on the growth and proliferation of colon cancer cells. Methods: Stable transfected colon cancer cell lines with RACK1 expression changes as well as corresponding control cell lines were established by using Lipofectamine 2000. Cell proliferative ability was analyzed by CCK-8 assay, soft agar colony formation assay, Ed U incorporation assay and flow cytometry. The expression of G1/S phase checkpoint regulation proteins Cyclin D1 and p27 was detected by Western blot. Results: Downregulation of RACK1 in colon cancer SW620 cells inhibited cell growth and soft agar colony formation ability, decreased number of S phase cells labeled by Ed U, and blocked cell cycle at G1/S phase. Overexpression of RACK1 in colon cancer SW480 cells enhanced cell growth and soft agar colony formation ability, increased number of S phase cells labeled by Ed U, and promoted progression of cell cycle G1/S phase. Conclusion: RACK1 overexpression promotes the growth and proliferation of colon cancer cells.