Disabled-1在人乳腺癌细胞中的表达及其对细胞周期的影响

doi:10.13481/j.1671-587x.20160518

吉林大学学报(医学版) ›› 2016, Vol. 42 ›› Issue (05): 932-936.doi: 10.13481/j.1671-587x.20160518

Disabled-1在人乳腺癌细胞中的表达及其对细胞周期的影响

曹让娟¹, 李凯², 邢琬莹³, 王悦书¹, 于维¹, 吴广智¹, 崔树森¹, 李强¹

1. 吉林大学中日联谊医院手外科, 吉林长春 130033;
2. 吉林大学中日联谊医院麻醉科, 吉林长春 130033;
3. 吉林大学中日联谊医院乳腺外科, 吉林长春 130033

收稿日期:2016-04-12 出版日期:2016-09-28 发布日期:2016-09-29
通讯作者: 李强,主治医师(Tel:0431-89876961,E-mail:alex334@163.com) E-mail:alex334@163.com
作者简介:曹让娟(1987-),女,重庆市人,讲师,医学博士,主要从事乳腺癌发生发展及周围神经损伤后再生等方面的研究。
基金资助:
国家自然科学基金资助课题（81502516）；吉林省科技厅科技发展计划项目资助课题（20160101077JC）；吉林省科技厅医药产业发展引导资金项目资助课题（20150311069YY）；吉林大学白求恩计划项目青年基金资助课题（2015429）

Expression of Disabled-1 in human breast cancer cells and its role in cell cycle

CAO Rangjuan¹, LI Kai², XING Wanying³, WANG Yueshu¹, YU Wei¹, WU Guangzhi¹, CUI Shusen¹, LI Qiang¹

1. Department of Hand Surgery, China-Japan Union Hospital, Jilin University, Changchun 130033;
2. Department of Anesthesiology, China-Japan Union Hospital, Jilin University, Changchun 130033;
3. Department of Breast Surgery, China-Japan Union Hospital, Jilin University, Changchun 130033, China

Received:2016-04-12 Online:2016-09-28 Published:2016-09-29

摘要/Abstract

摘要：

目的：探讨Disabled-1（Dab1）基因在乳腺上皮细胞及乳腺癌细胞中的表达情况，并阐明Dab1对乳腺癌细胞周期的影响。方法：体外培养永生化的乳腺上皮细胞MCF-10A及乳腺癌细胞MCF-7、BT-549和MDA-MB-231，实时荧光定量（Real-time PCR）法检测MCF-10A、MCF-7、BT-549和MDA-MB-231细胞中Dab1 mRNA的表达水平，Western blotting法检测Dab1蛋白的表达水平。取对数生长期的BT-549细胞，分为对照组、转染pKH3组和转染pKH3-Dab1组，采用流式细胞术检测各组细胞周期的变化情况。结果：Real-time PCR法检测，与MCF-10A细胞（0.998±0.020）比较，乳腺癌细胞MCF-7（0.504±0.037）、BT-549（0.302±0.027）和MDA-MB-231（0.330±0.031）中Dab1 mRNA相对表达水平明显下降（P<0.05）。Western blotting法检测，与MCF-10A细胞（0.227±0.021）比较，乳腺癌细胞MCF-7（0.134±0.014）、BT-549（0.076±0.01）和MDA-MB-231（0.074±0.005）中Dab1蛋白表达水平明显下调（P<0.05）。流式细胞术检测，与对照组和转染pKH3组比较，转染pHK3-Dab1组BT-549细胞出现细胞周期G₁期阻滞。结论：Dab1在体外培养乳腺癌细胞系中表达下调，Dab1过表达可阻滞乳腺癌细胞G₁细胞周期进展。

关键词: Dab1基因, 人乳腺癌细胞, 细胞周期, G₁期阻滞

Abstract:

Objective: To explore the expressions of Disabled-1(Dab1) in human breast epithelial cells and breast cancer cells, and to clarify its role in cell cycle. Methods: Real-time PCR was used to analyze the Dab1 mRNA expressions in breast epithelial cells MCF-10A and breast cancer cells MCF-7, BT-549, and MDA-MB-231. The Dab1 protein expressions in those cells were tested by Western blotting method. The BT-549 cells at logarithmic growth phase were divided into control, pKH3, and pKH3-Dab1 groups; the cell cycle was investigated by flow cytometry. Results:The Real-time PCR results showed that the Dab1 mRNA expression levels in MCF-7 cells (0.504±0.037), BT-549 cells (0.302±0.027), and MDA-MB-231 cells (0.330±0.031) were reduced compared with MCF-10A cells (0.998±0.020) (P<0.05). The Western blotting results showed that the Dab1 protein expression levels in breast cancer cells MCF-7 (0.134±0.014), BT-549 (0.076±0.01), and MDA-MB-231 (0.074±0.005) were reduced compared with MCF-10A cells (0.227±0.021) (P<0.05). Compared with control group and pKH3 group, the cell cycle in pKH3-Dab1 group was inhibited at G₁ phase detected by flow cytometry analysis. Conclusion: The expression of Dab1 is down-regulated in breast cancer cells, and the over-expression of Dab1 can inhibit the cell cycle at G1 phase.

Key words: Dab1, human breast cancer cell, cell cycle, G₁ phase inhibition

中图分类号:

R730.2

曹让娟, 李凯, 邢琬莹, 王悦书, 于维, 吴广智, 崔树森, 李强. Disabled-1在人乳腺癌细胞中的表达及其对细胞周期的影响[J]. 吉林大学学报(医学版), 2016, 42(05): 932-936.

CAO Rangjuan, LI Kai, XING Wanying, WANG Yueshu, YU Wei, WU Guangzhi, CUI Shusen, LI Qiang. Expression of Disabled-1 in human breast cancer cells and its role in cell cycle[J]. Journal of Jilin University Medicine Edition, 2016, 42(05): 932-936.

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Disabled-1在人乳腺癌细胞中的表达及其对细胞周期的影响

Expression of Disabled-1 in human breast cancer cells and its role in cell cycle

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