【摘要】：目的觀察Hedgehog通路抑制劑GANT61對(duì)人乳腺癌MCF-7細(xì)胞的抑制作用。方法 GANT61作用MCF-7細(xì)胞后,通過流式細(xì)胞術(shù)檢測(cè)細(xì)胞死亡;提取處理細(xì)胞總RNA和蛋白。分別采用Real-time PCR和Western blot檢測(cè)Hedgehog通路SHH、Gli1、Gli2及自噬標(biāo)志物L(fēng)C3-Ⅱ表達(dá);處理細(xì)胞固定后通過電子顯微鏡觀察GANT61誘導(dǎo)的自噬體形態(tài);自噬抑制劑3-MA處理細(xì)胞或自噬相關(guān)基因atg5 siRNA轉(zhuǎn)染細(xì)胞后再用GANT61作用,流式細(xì)胞術(shù)檢測(cè)細(xì)胞死亡數(shù)量。結(jié)果 GANT61能明顯誘導(dǎo)MCF-7死亡,降低Hedgehog通路SHH、Gli1、Gli2表達(dá)水平,提高LC3-Ⅱ蛋白表達(dá),并可誘導(dǎo)MCF-7細(xì)胞產(chǎn)生自噬體。3-MA及atg5 siRNA可減弱GANT61誘導(dǎo)的細(xì)胞死亡。結(jié)論 GANT61通過誘導(dǎo)自噬性細(xì)胞死亡機(jī)制發(fā)揮抗乳腺癌細(xì)胞活性。
[Abstract]:Objective To observe the inhibitory effect of the Hedgehog pathway inhibitor, GANT61, on human breast cancer MCF-7 cells. Methods The cell death was detected by flow cytometry, and the total RNA and protein of the cells were extracted by flow cytometry. The expression of the Hedgehog pathway SHH, Gli1, Gli2 and the self-autophagy marker LC3-鈪，

本文編號(hào)：2412616