微信公众号

官网二维码

中国癌症防治杂志 ›› 2018, Vol. 10 ›› Issue (3): 188-192.doi: 10.3969/j.issn.1674-5671.2018.03.05

• 乳腺癌专栏 • 上一篇    下一篇

过表达RSK4m1对人乳腺癌细胞MDA-MB-231生长和侵袭的影响

  

  1. 广西医科大学附属肿瘤医院乳腺一科;广西医科大学研究生院
  • 出版日期:2018-06-25 发布日期:2018-07-02
  • 通讯作者: 杨华伟 E-mail:Lordyhw@163.com
  • 基金资助:

    国家自然科学基金资助项目(81260394)

Effect of overexpression of ribosomal protein S6 kinase 4 variants 1 on proliferation and invasion by human breast cancer cell MDA-MB-231

  • Online:2018-06-25 Published:2018-07-02

摘要:

目的 探讨过表达p90核糖体S6蛋白激酶4变异体1(ribosomal protein S6 kinase 4 variant 1,RSK4m1)对人乳腺癌MDA-MB-231细胞生长和侵袭的影响。方法 通过负载RSK4m1慢病毒在人乳腺癌MDA-MB-231细胞中稳定过表达RSK4m1。以MDA-MB-231亲本细胞为空白对照组(Con组)、转染空载病毒的MDA-MB-231细胞为阴性对照组(Mock组)、转染过表达RSK4m1的MDA-MB-231细胞为实验组(OE组)。采用qRT-PCR和Westen Blot法检测各组RSK4m1 mRNA及蛋白的表达;CCK-8法检测细胞增殖情况;细胞划痕实验检测细胞的迁移能力;Transwell小室侵袭实验检测细胞的侵袭能力。结果 OE组中RSK4m1 mRNA及蛋白的表达量均明显高于Con组及Mock组(P<0.05)。CCK-8实验显示,24 h、48 h、72 h和96 h时OE组细胞增殖均低于Mock组和Con组(P<0.05);细胞划痕实验显示,细胞培养24 h后,OE组细胞迁移率显著低于Con组和Mock组[(14.53±0.64)% vs (25.67±2.44)%、(24.47±2.25)%,P<0.05]。Transwell小室侵袭实验显示,OE组细胞侵袭能力显著低于Con组和Mock组[(35.00±5.57)个 vs (66.70±5.86)个、(58.67±4.16)个,P<0.05]。结论 过表达RSK4m1可显著抑制人乳腺癌MDA-MB-231细胞增殖、迁移和侵袭能力,为RSK4m1可能成为乳腺癌的治疗新靶点提供了实验依据。

关键词: 乳腺肿瘤, 人乳腺癌细胞株, RSK4m1, 增殖, 侵袭, 迁移

Abstract:

Objective To investigate the effect of ribosomal protein S6 kinase 4 variants 1 (RSK4m1) on the proliferation and invasion by human breast cancer cell line MDA-MB-231. Methods RSK4m1 was stably overexpressed in human breast cancer cell line MDA-MB-231 using lentivirus encoding RSK4m1. Untreated MDA-MB-231 cells served as a blank control group,MDA-MB-231 cells transfected with empty virus served as a negative control group and MDA-MB-231 cells transfected with the RSK4m1 overexpression cassette served as the experimental group. The expression of RSK4m1 in each group was detected by real-time PCR and Western blot analysis. The proliferation of cells was assayed using the cell counting kit-8 (CCK-8),and cell migration was measured using the wound healing test. Invasion was detected using the Transwell test. Results The mRNA and protein expression of RSK4m1 was significantly up-regulated in the experimental group(P<0.05). The CCK-8 assay showed that the proliferation of the experimental group was significantly lower than that of the negative control group and blank control group at 24,48,72 and 96 h(P<0.05). The wound healing test showed that the closure rate of the cells in the experimental group was significantly lower than that in the blank control group and the negative control group after 24 h of cell culture(P<0.05). The transwell test showed that the invasive ability of the experimental group was significantly lower than that in the blank control group and the negative control group (P<0.05). Conclusions Overexpression of RSK4 variant 1 can effectively suppress cell proliferation,migration and invasion by MDA-MB-231 cells,which provides an experimental basis for researching RSK4 variant 1 as a new target in the treatment of human breast cancer.

Key words: Breast neoplasms, Human breast cell lines, Ribosomal protein S6 kinase 4 variants 1, Cell proliferation, Cell invasion, Cell migration