【作者】 姚舒洋；

【导师】 徐兵河； 詹启敏； 宋咏梅；

【作者基本信息】 中国协和医科大学 ， 肿瘤学， 2010， 博士

【摘要】 三阴性乳腺癌(TNBC)约占新诊断乳腺癌的15%,这类患者发病年龄早,易早期局部复发和远处转移,无病生存时间和总生存时间均短,预后不佳,目前国内外仍缺乏针对这类特殊类型乳腺癌的规范化治疗指南。本课题旨在研究索拉非尼(Sorafenib)联合顺铂(DDP)对TNBC细胞生长的抑制作用,进一步从信号通路、细胞周期和细胞凋亡等方面探讨了Sorafenib联合顺铂(DDP)治疗TNBC的可能机制。MTT法检测发现单药DDP和Sorafenib在体外对TNBC细胞株MDA-MB-231、MDA-MB-468、CAL-51和非TNBC细胞株MCF-7的增殖都有一定的抑制作用。与非TNBC细胞MCF-7相比,MDA-MB-468和CAL-51细胞对DDP更为敏感。通过中效原理,证实DDP和Sorafenib联合应用时,在各个细胞株中均为协同效应。DDP联合Sorafenib对各细胞株协同抑制的分子机制主要是通过影响MAPK通路关键蛋白的表达实现的,Western blot检测显示各细胞株均表现为p-ERK和P38蛋白表达水平不同程度的下降以及p-JNK和p-P38蛋白表达水平不同程度的升高。流式细胞术检测发现DDP联合Sorafenib能阻滞各细胞株的细胞周期,使细胞阻滞在G0／G1期,而抑制细胞增殖。Western blot检测在蛋白水平发现CyclinA、CylinB1、CylinD1和CylinE多种细胞周期蛋白明显降低,这可能是细胞周期抑制的原因。DDP联合Sorafenib诱导各细胞株凋亡的机制可能主要是通过抑制Bc1-2蛋白的表达实现的。本研究的意义在于首次发现Sorafenib联合DDP在体外能协同抑制多种TNBC细胞株的增殖,并初步阐明分子机制,为该方案的临床应用提供了理论依据。更多还原

【Abstract】 Triple-negative breast cancer (TNBC) accounts for approximately 15% of breast cancer diagnoses. Although triple-negative breast cancer accounts for a relatively small minority of breast cancer cases, it is responsible for a disproportionate number of breast cancer deaths. Meanwhile, this disease often occurs in young women. Multiple data sets have consistently identified poorer clinical outcomes for women with TNBC. Until recently, the subset of breast cancer lacking the expressions of estrogen receptor, progesterone receptor and human epithelial growth factor receptor 2 had no standard therapeutic approach. This study is to investigate the inhibitory proliferation effect of Sorafenib combined with cisplatin (DDP) on TNBC cells and to identify the possible molecular mechanisms by detecting the molecules associated with MAPK signaling pathway, cell cycle and apoptosis.The results of MTT showed that Sorafenib or DDP alone inhibited TNBC cell lines MDA-MB-231, MDA-MB-468, CAL-51 and non-TNBC cell line MCF-7 proliferation dose-dependently in vitro. Compared with non-TNBC cell line MCF-7, TNBC cell lines MDA-MB-468 and CAL-51 were more sensitive to DDP. Based on the median-effect principle, Sorafenib combined with DDP showed a synergistic inhititory effect on the four cell lines proliferation. The potential molecular machenism of Sorafenib with DDP on synergistic inhititory effect was closely related to the blockage or activation of MAPK signling pathways. Western blot analysis indicated p-ERK and P38 protein expression levels decreased, but p-JNK and p-P38 protein expression levels elevated in four cell lines with different degrees. Sorafenib combined with DDP blocked cell cycle progress by Flow Cytometry. Sorafenib with DDP arrested cells in G0/G1 phase and inhibited cell proliferation. Western blot analysis also indicated that decreased expression of CyclinA, CylinB1, CylinD1 and CylinE in the four cell lines would be the reason of cell cycle blockage. Sorafenib with DDP induced cell apoptosis possiblely by inhibiting the expression of Bcl-2 protein.This study suggested that Sorafenib combined with DDP had a synergistic inhibitory effect on TNBC cells proliferation. The mechanisms of synergistic effect were closely related to the regulation on MAPK signaling pathway, blockage of cell cycle progress and the enhancement of apoptosis.Therefore, the findings in this report provide insights into coadministration of Sorafenib and DPP as a regimen in clinical practice in treating TNBC patients.更多还原