RACK1高表达促进人结肠癌细胞的生长和增殖
| 作者: |
1曹超,
1黄伟,
1肖志强,
1易红
1 中南大学湘雅医院肿瘤蛋白质组转化医学湖南省高校重点实验室，长沙 410008 |
| 通讯: |
易红
Email: yh1126@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2016.04.021 |
| 基金: | 国家973计划课题, 2013CB910502 国家自然科学基金, 81172302 |
摘要
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RACK1表达下调的SW620细胞系、RACK1表达上调的SW480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、EdU掺入实验以及流式细胞术检测RACK1表达改变对SW620和SW480细胞增殖的影响；采用Western blot分析RACK1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EdU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SW480细胞的生长、软琼脂集落形成能力，增加EdU标记的S期细胞数目，促进细胞周期G1/S期进程。结论：RACK1高表达促进人结肠癌细胞的生长和增殖。
关键词：
RACK1
结肠癌
细胞生长
细胞增殖
RACK1 overexpression promotes the growth and proliferation of colon cancer cells
CorrespondingAuthor: YI Hong Email: yh1126@163.com
DOI: 10.3978/j.issn.2095-6959.2016.04.021
Abstract
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, EdU 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 EdU, 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 EdU, and promoted progression of cell cycle G1/S phase. Conclusion: RACK1 overexpression promotes the growth and proliferation of colon cancer cells.