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

作　　者：宋扬 刘佩娟[2] 辛智倩 张波[3] 赵勇[2] 张彩勤[2] 白冰[2] 师长宏[2] 张海[2] 

机构地区：[1]解放军323医院口腔科,西安710054 [2]第四军医大学实验动物中心,西安710032 [3]第四军医大学唐都医院泌尿外科,西安710038

出　　处：《肿瘤防治研究》2017年第1期1-4,共4页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金面上项目(31272391);陕西省科技资源统筹项目(2014FWPT-11)

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。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 m RNA 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.

关 键 词：GANT61 自噬性细胞死亡 乳腺癌 MCF-7细胞 

分 类 号：R737.9[医药卫生—肿瘤]