Effect of the tumor suppressor gene ING4 on the proliferation of MCF-7 human breast cancer cells.
Effect of the tumor suppressor gene ING4 on the proliferation of MCF-7 human breast cancer cells.
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DOI：
10.3892/ol.2012.744
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发表时间：
2012-09
期刊：
影响因子：
2.9
通讯作者：
Qinjun Wei;Wei He;Ya-jie Lu;Jun Yao;Xin Cao
中科院分区：
文献类型：
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作者：
Qinjun Wei;Wei He;Ya-jie Lu;Jun Yao;Xin Cao
生长抑制剂4 （Inhibitor of growth 4, ING4）是ING家族的一员，是一种肿瘤抑制蛋白。为了研究ING4对乳腺癌增殖的影响，本研究在MCF-7细胞中检测了ING4表达上调引起的抗肿瘤作用及其可能的作用机制。利用编码ING4基因的质粒表达系统构建稳定细胞系，并在MCF-7细胞中过表达ING4。采用Real-time PCR和western blot分别检测ING4 mRNA和蛋白的表达水平。采用甲基噻唑四氮唑（MTT）法检测细胞生长情况。流式细胞术检测细胞周期分布和细胞凋亡情况。采用实时荧光定量PCR和western blot检测p21、p53和bax基因的表达。建立了稳定转染的细胞系pcDNA3.1(+)/ING4（含ING4基因）和pcDNA3.1(+)（空载体）。在稳定表达pcDNA3.1(+)/ING4的细胞系中，ING4 mRNA和蛋白的表达水平显著高于对照细胞系。稳定转染后的细胞增殖受到抑制，抑制率为62.58±2.93%。从稳定转染细胞与对照细胞系细胞周期分布的变化来看，稳定转染细胞系中有67.82±3.78%的细胞被阻断在G0/G1期（P0.05）。p21、bax和p53的Western blot分析结果与real-time PCR结果一致。ING4上调可能抑制乳腺癌细胞增殖，加速细胞凋亡过程。提示ING4在抑制乳腺癌进展中起重要作用。
Inhibitor of growth 4 (ING4) is a member of the ING family and acts as a tumor suppressor protein. To investigate the impact of ING4 on breast cancer proliferation, the present study examined the antitumor effects caused by upregulation in the expression of ING4 and its possible mechanism of effect in MCF-7 cells. A plasmid-based expression system encoding the ING4 gene was used to construct a stable cell line and overexpress ING4 in MCF-7 cells. Real-time PCR and western blot analysis were used to detect the mRNA and protein expression levels of ING4, respectively. Cell growth was examined by methylthiazolyltetrazolium (MTT) assay. Cell cycle distribution and cell apoptosis were measured by flow cytometry. The expression of p21, p53 and bax genes were tested by real-time PCR and western blot analysis. The stably transfected cell lines pcDNA3.1(+)/ING4 (with the ING4 gene) and pcDNA3.1(+) (an empty vector) were established. The expression levels of ING4 mRNA and protein in the stable cell line expressing pcDNA3.1(+)/ING4 were significantly higher than those of the two control cell lines. The cell proliferation of stably transfected cells was inhibited, and the inhibitory rate was 62.58±2.93%. Based on the changes in cell cycle distribution in stably transfected cells compared with two control cell lines, a number of cells were blocked in the G0/G1 phase 67.82±3.78% (P0.05) in the stably transfected cell lines. Western blot analysis results of p21, bax and p53 were in accordance with real-time PCR results. ING4 upregulation may inhibit breast cancer cell proliferation and accelerate the process of apoptosis. It is suggested that ING4 plays a significant role in the suppression of breast cancer progression.