[转染pIRES-p21（waf1）-p27（kip1）基因对乳腺癌细胞系MCF-7中心体复制及增殖的影响]

[Effects of transfected pIRES-p21(waf1)-p27(kip1) gene on the duplication of centrosomes and the proliferation of breast cancer cell line MCF-7].

作者信息

Jiang Dan-dan, Li Fu-nian, Liu Xiang-ping

机构信息

Department of Breast Surgary, the Affiliated Hospital of Qingdao Medical College, Qingdao 266003, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2009 Dec 15;89(46):3295-8.

PMID:20193371

Abstract

OBJECTIVE

To explore the role of transfected pIRES-p21(waf1)-p27(kip1) gene on the centrosome duplication and cell proliferation of MCF-7, a breast cancer cell line.

METHODS

The pIRES-p21(waf1), pIRES-p27(kip1) and pIRES-p21(waf1)-p27(kip1) genes were transfected into the MCF-7 cells by lipofection. The effect on proliferation was evaluated by MTT assay and cell growth curve was drawn. The cell cycle was analyzed by flow cytometry. The centrosome duplication was detected by using indirect immunofluorescence microscopy.

RESULTS

After transfected 24 hours, the p21(waf1) and p27(kip1) protein expressions were significantly increased as compared with untransfected MCF-7 cells (P < 0.01), and cell growth was obviously inhibited and resulted in an accumulation of cells in G(1) (P < 0.01), presenting that the proportion of cells in G(1) phase was obviously increased from(47.28 +/- 2.25)% to (69.52 +/- 3.21)% of p21(waf1) transfected cells, (60.83 +/- 3.02)% of p27(kip1) transfected cells, and (78.37 +/- 2.83)% of p21 (waf1)-p27(kip1) transfected cells. The proportion of cells which contained unnormal centrosomes was obviously decreased, from (13.47 +/- 0.33)% to (5.07 +/- 0.38)%, (6.28 +/- 0.35)%, (3.47 +/- 0.23)%, respectively.

CONCLUSION

The transfer of p21(waf1) and p27(kip1) genes could inhibit the growth of human breast carcinoma cells and the unnormal duplication of centrosomes. p21(waf1) had a really synergy with p27(kip1) in these effects, suggesting p21(waf1)-p27(kip1) combined gene can inhibit the genesis and development of breast cancer and might have potential clinical significance as therapeutic agents of breast cancer.

摘要

目的

探讨转染pIRES-p21（waf1）-p27（kip1）基因对乳腺癌细胞系MCF-7中心体复制及细胞增殖的作用。

方法

采用脂质体转染法将pIRES-p21（waf1）、pIRES-p27（kip1）和pIRES-p21（waf1）-p27（kip1）基因转染至MCF-7细胞。通过MTT法评估对增殖的影响并绘制细胞生长曲线。采用流式细胞术分析细胞周期。利用间接免疫荧光显微镜检测中心体复制情况。

结果

转染24小时后，与未转染的MCF-7细胞相比，p21（waf1）和p27（kip1）蛋白表达显著增加（P<0.01），细胞生长明显受抑制并导致细胞在G（1）期积累（P<0.01），表现为G（1）期细胞比例明显增加，p21（waf1）转染细胞从（47.28±2.25）%增至（69.52±3.21）%，p27（kip1）转染细胞从（60.83±3.02）%增至（78.37±2.83）%，p21（waf1）-p27（kip1）转染细胞从（60.83±3.02）%增至（78.37±2.83）%。含有异常中心体的细胞比例明显降低，分别从（13.47±0.33）%降至（5.07±0.38）%、（6.28±0.35）%、（3.47±0.23）%。