PPT诱导高表达人源HER2乳腺癌细胞凋亡及相关机制研究
发布时间：2019-02-13 08:36
【摘要】：本研究首先以4株人源乳腺癌细胞MCF7、MDA-MB-231、MDA-MB-453和SK-BR-3为基础,通过大量MTT实验确定鬼臼毒素作为典型的体外抗乳腺癌天然药物。分析比较鬼臼毒素对各人源乳腺癌细胞的IC50值,确认出对鬼臼毒素较为敏感细胞株MDA-MB-453和SK-BR-3。Hoechst染色法和流式细胞术检测鬼臼毒素诱导高表达HER2阳性乳腺癌细胞凋亡的能力,Western blot和实时荧光定量PCR实验分别检测鬼臼毒素作用后高表达HER2阳性乳腺癌细胞中Cleaved-caspase-3、Cleaved-caspase-9、Bcl-2Bax等凋亡相关蛋白和mRNA的表达水平。结果显示,相比5FU和大黄素,鬼臼毒素对多种人源乳腺癌细胞增殖的抑制作用更显著(P0.05),可以作为典型的体外抗乳腺癌天然药物。相比于HER2阴性乳腺癌细胞MCF7和MDA-MB-231,鬼臼毒素作用下高表达HER2阳性乳腺癌细胞MDA-MB-453和SK-BR-3都发生了明显凋亡(P0.05);其机制可能是一方面通过激活Cleaved-caspase-3、Cleaved-caspase-9,另一方面通过上调Bax,下调Bcl-2激活线粒体途径来诱导高表达HER2阳性乳腺癌细胞凋亡(P0.05)。
[Abstract]:In this study, based on four human breast cancer cell lines MCF7,MDA-MB-231,MDA-MB-453 and SK-BR-3, podophyllotoxin was identified as a typical natural anti-breast cancer drug by a large number of MTT experiments. The IC50 values of podophyllotoxin on human breast cancer cells were analyzed and compared. To confirm the ability of podophyllotoxin sensitive cell line MDA-MB-453 and SK-BR-3.Hoechst staining and flow cytometry to detect apoptosis induced by podophyllotoxin in high expression HER2 positive breast cancer cells. Western blot and real-time fluorescence quantitative PCR assay were used to detect the expression of Cleaved-caspase-3,Cleaved-caspase-9,Bcl-2Bax and mRNA in HER2 positive breast cancer cells after podophyllotoxin treatment. The results showed that podophyllotoxin could inhibit the proliferation of human breast cancer cells more significantly than 5FU and emodin (P0.05) and could be used as a typical anti-breast cancer natural drug in vitro. Compared with HER2 negative breast cancer cells MCF7 and MDA-MB-231, podophyllotoxin high expression of HER2 positive breast cancer cells MDA-MB-453 and SK-BR-3 were significantly apoptotic (P0.05). On the one hand, the mechanism may be by activating Cleaved-caspase-3,Cleaved-caspase-9, on the other hand, by up-regulating Bax, down-regulating Bcl-2 and activating mitochondrial pathway to induce apoptosis of high expression HER2 positive breast cancer cells (P0.05).
【学位授予单位】：华东理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9
本文编号：2421366
[Abstract]:In this study, based on four human breast cancer cell lines MCF7,MDA-MB-231,MDA-MB-453 and SK-BR-3, podophyllotoxin was identified as a typical natural anti-breast cancer drug by a large number of MTT experiments. The IC50 values of podophyllotoxin on human breast cancer cells were analyzed and compared. To confirm the ability of podophyllotoxin sensitive cell line MDA-MB-453 and SK-BR-3.Hoechst staining and flow cytometry to detect apoptosis induced by podophyllotoxin in high expression HER2 positive breast cancer cells. Western blot and real-time fluorescence quantitative PCR assay were used to detect the expression of Cleaved-caspase-3,Cleaved-caspase-9,Bcl-2Bax and mRNA in HER2 positive breast cancer cells after podophyllotoxin treatment. The results showed that podophyllotoxin could inhibit the proliferation of human breast cancer cells more significantly than 5FU and emodin (P0.05) and could be used as a typical anti-breast cancer natural drug in vitro. Compared with HER2 negative breast cancer cells MCF7 and MDA-MB-231, podophyllotoxin high expression of HER2 positive breast cancer cells MDA-MB-453 and SK-BR-3 were significantly apoptotic (P0.05). On the one hand, the mechanism may be by activating Cleaved-caspase-3,Cleaved-caspase-9, on the other hand, by up-regulating Bax, down-regulating Bcl-2 and activating mitochondrial pathway to induce apoptosis of high expression HER2 positive breast cancer cells (P0.05).
【学位授予单位】：华东理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9
【参考文献】
相关期刊论文 前1条
1 田崛;陈显凌;庄英婷;范莹娟;许建华;吴丽贤;;BMS-345541对急性粒细胞白血病细胞DNA损伤修复的影响[J];中国药理学通报;2015年06期
，本文编号：2421366
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