大黄酸对乳腺癌肿瘤细胞HER-2蛋白表达的影响研究

Effects of Rhein on Expression of HER-2 Protein in Breast Cancer Cells

目的:对大黄酸用于乳腺癌治疗中对肿瘤细胞的HER-2蛋白表达的影响进行研究,证实大黄酸在乳腺癌治疗中的价值及作用机制,为临床及研究提供参考。方法:64例乳腺癌患者,提取相应的细胞样本,采用MTT法对肿瘤的细胞增殖情况、受体PT心通路蛋白表达水平、磷酸化水平的变化进行检测;使用免疫细胞化学检测法对EGFR和HER-2蛋白表达水平进行检测;HER-2在基因转录水平上的变化使用RT-PCR检测,细胞凋亡和细胞周期使用流式细胞技术(FACS)检测。收集检测结果并进行数据分析。结果:(1)大黄酸对MCF-7、SK-BR-3和MDA.MB-231等乳腺癌细胞、卵巢癌SK-OV-3细胞系和人脐静脉内皮细胞(HUVEC)均有一定的抑制作用,其半抑制率IC50分别为98μmol/L、86μmol/L、115μmol/L、88μmol/L和205μmol/L。(2)大黄酸的抗肿瘤机制与RHL相似,均是通过1通过抑制EGFR的磷酸化及其下游Raf/MEK/ERK信号通路的磷酸化抑制EGFR高表达和HER-2低表达的MCF-7细胞增殖;2对于EGFR和HER-2高表达的SK-Bf-3细胞,则通过抑制EGFR和HER-2蛋白表达和蛋白磷酸化水平,同时诱导肿瘤细胞通过p53和p21途径发生凋亡,两种方式发挥作用的。结论:大黄酸对HER-2双靶点受体蛋白酪氨酸激酶具有靶向抑制作用,能够通过抑制细胞增殖和诱导细胞凋亡两种途径发挥其抗乳腺癌肿瘤的细胞作用,进而达到治疗乳腺癌,改善患者生活质量的目的。

Objective： To study the effect of Rhein on tumor cells for expression of HER -2 protein in the treatment ofbreast cancer and to confirm the value and the function mechanism of rhein in breast cancer treatment and provide refer-ence for clinical and research. Methods： 64 cases of breast cancer patients in our hospital in 2012 March-March 2014were as the objects of study. The change of the expression of tumor cell proliferation, PT receptor protein levels and heartpathway phosphorylation were detected by MTT method. Levels of EGFR and HER - 2 protein were detected by using im-munocytochemical detection method. HER - 2 changes in gene transcription level were detected by using RT - PCR as-say. Cell apoptosis and cell cycle were detected by flow cytometry （FACS） detection. We collected test results and madethe data analysis. Results ： （ 1 ） Rhein can inhibit MCF. 7 and SK - BR - 3 and MDA. MB - 231 of breast cancer and ovar-ian cancer cell line SK. OV. 3 and human umbilical vein endothelial cells （HUVEC）. The ICs0 values were respectively98 μmol/L,86 μmol/L, 115 μmol/L,88μmol/L and 205 μmol/L. （2） Rhein anti - tumor mechanism was similar tothat of RHL. Both can inhibit the expression of EGFR and the low expression of HER - 2 through the inhibition of thephosphorylation of EGFR and its downstream Raf/MEK/ERK pathway phosphorylation. As for the EGFR and HER - 2expression in SK - Bf - 3 cells, both are through the inhibition of EGFR and HER - 2 protein expression and protein phos-phorylation to induce tumor cell apoptosis by P53 and p21 pathways. Conclusion： Rhein has targeted inhibition of EGFRand HER -2 double target receptor protein tyrosine kinase and can inhibit breast cancer through two ways to inhibit cellproliferation and induce cell apoptosis,and then to treat breast cancer and improve the quality of life of the patients.