GANT61通过自噬性细胞死亡抑制人乳腺癌MCF-7细胞增殖

宋扬; 刘佩娟; 辛智倩; 张波; 赵勇; 张彩勤; 白冰; 师长宏; 张海

doi:10.3971/j.issn.1000-8578.2017.01.001

GANT61通过自噬性细胞死亡抑制人乳腺癌MCF-7细胞增殖

Autophagic Cell Death Induced by GANT61 Inhibits Proliferation of Human Breast Cancer Cell Line MCF-7

摘要

摘要:
目的观察Hedgehog通路抑制剂GANT61对人乳腺癌MCF-7细胞的抑制作用。
方法 GANT61作用MCF-7细胞后，通过流式细胞术检测细胞死亡；提取处理细胞总RNA和蛋白。分别采用Real-time PCR和Western blot检测Hedgehog通路SHH、Gli1、Gli2及自噬标志物LC3-Ⅱ表达；处理细胞固定后通过电子显微镜观察GANT61诱导的自噬体形态；自噬抑制剂3-MA处理细胞或自噬相关基因atg5 siRNA转染细胞后再用GANT61作用，流式细胞术检测细胞死亡数量。
结果 GANT61能明显诱导MCF-7死亡，降低Hedgehog通路SHH、Gli1、Gli2表达水平，提高LC3-Ⅱ蛋白表达，并可诱导MCF-7细胞产生自噬体。3-MA及atg5 siRNA可减弱GANT61诱导的细胞死亡。
结论 GANT61通过诱导自噬性细胞死亡机制发挥抗乳腺癌细胞活性。

Abstract:
Objective To investigate the inhibitory effect of GANT61 on human breast cancer cell line MCF-7.
Methods Cell death was measured by flow cytometry after GANT61 treatment; the total RNA and protein were extracted from the treated cells; the mRNA and protein expression levels of SHH, Gli1, Gli2 and LC3-Ⅱ were detected by Real-time PCR and Western blot, respectively. Autophagosomes formation was observed under transmission electronic microscopy (TEM). After that, 3-MA and atg5 siRNA were used to illustrate the relationship between autophagy and cell death.
Results GANT61 induced significantly the cell death of MCF-7 cells. The expressions of SHH, Gli1 and Gli2 were decreased; however, autophagy marker LC3-II expression was increased after GANT61 treatment. Meanwhile, autophagosomes could be observed under TEM. 3-MA and atg5 siRNA could inhibit GANT61-induced cell death.
Conclusion GANT61 inhibits the proliferation of human breast cancer cell line MCF-7 through autophagic cell death.

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