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Smad1参与调节雌激素在乳腺癌细胞中的促增殖作用

Smad1 involves in regulating proliferation effect of estrogen in breast cancer cells

  • 摘要: 目的: 观察在雌激素促进乳腺癌发生、发展的过程中,Smad1与雌激素的相互作用关系,以及其对乳腺癌细胞增殖、侵袭功能的影响,并探讨其作用机制。方法: 通过Western印迹法以及实时定量聚合酶链式反应(real-time PCR)技术检测雌激素作用与Smad1表达的相关关系;采用小发卡RNA(short hairpin RNA,shRNA)干扰技术下调乳腺癌细胞中Smad1表达,并检测其对雌激素作用的影响。通过Transwell法、免疫组化法(IHC)、细胞计数法等研究Smad1表达情况对乳腺癌细胞增殖、侵袭能力的影响。结果: Smad1能够在雌激素刺激下提高其转录和翻译效应,雌激素作用4 h后磷酸化Smad-1(p-Smad1)被激活,并在12 h达到最大值(P< 0.001)。乳腺癌肿瘤组织中Smad1的表达强于癌旁组织,且Smad1水平与雌激素受体α(ERα)正相关,而p-Smad1的活化被抑制。应用RNA干扰技术沉默两种乳腺癌细胞中Smad1后,细胞体外增殖速度显著下降(P< 0.05);在体内试验中,肿瘤的生长也受到显著抑制(P< 0.05)。结论: 雌激素对乳腺癌细胞的促增殖和迁移作用依赖于Smad1;骨形成蛋白(bone morphogenetic protein, BMP)-Smad1途径在雌激素促乳腺癌发生和发展过程中可能发挥重要的调节作用。

     

    Abstract: Objective: To investigate the interaction between Smad1 and estrogen in the promotion of breast cancer, as well as its functional role and underlying mechanism. Methods: The interaction relationship of Smad1 and estrogen was detected by Western blotting and real-time PCR. Smad1 was down-regulated using short hairpin RNA (shRNA) interference technology in breast cancer cells. Using Transwell method, IHC and cell counting method to observe the effect of Smad1 on cell proliferation and invasion. Results: Smad1 could increase transcription and translation of breast cancer cells in the stimulation of estrogen. p-Smad1 was activated after four hour activation of estrogen and then reach maximum at twelve hour later (P< 0.001). The expression of Smad1 in cancer tissue was enhanced compared to paratumor tissues and Smad1 level was positively correlated with ERα while p-Smad1 activation was inhibited. In vitro cell proliferation, RNA interference silencing of Smad1 in two breast cancer cells decreased propagation ability significantly (P< 0.05); in vivo tests, tumor growth was also significantly inhibited (P< 0.05). Conclusions: The study indicates estrogen manipulation in breast cancer cells depending on Smad1. BMP-Smad1 pathway plays complex controlling role in the process of cell occurrence and development.

     

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