硫利达嗪对乳腺癌细胞MDA-MB-231和MCF-7的杀伤效应

发布时间：2018-05-25 14:45

本文选题：乳腺癌 + 硫利达嗪　；参考：《安徽医科大学》2015年硕士论文

【摘要】：背景与目的乳腺癌是女性最常见的恶性肿瘤之一,其发病率逐年上升且趋向低龄化,尤以ER、PR及Her-2表达阴性的三阴乳腺癌恶性程度更高,预后较差,如何有效针对三阴乳腺癌的新药研究成为目前乳腺癌治疗的重点[1-2]。硫利达嗪为吩噻嗪衍生物,其被广泛用于治疗严重的精神和情绪障碍[3]。最新研究[4]证实硫利达嗪在体外可对抗多种肿瘤细胞活性,提示其可能作为一种新型抗癌药物应用于以后的肿瘤临床治疗。该研究以硫利达嗪作用于三阴乳腺癌MDA-MB-231细胞和乳腺癌MCF-7细胞,观察硫利达嗪对人乳腺癌细胞MDA-MB-231和MCF-7的凋亡作用,并探讨其机制。方法采用四甲基偶氮唑蓝(MTT)法测定硫利达嗪对细胞的抑制作用,计算半数抑制浓度(IC50)。流式细胞术检测硫利达嗪对细胞周期分布和凋亡的影响。比色法测定药物对细胞Caspase-3活性的影响,Western blot法检测凋亡调节蛋白Bcl-2,Bax表达的变化。结果硫利达嗪作用24 h后,MDA-MB-231、MCF-7细胞增殖明显受到剂量依赖性抑制,其IC50分别为18、22μmol/L。流式细胞术结果显示,随着加入硫利达嗪浓度的提高,MDA-MB-231、MCF-7细胞均发生不同程度的G0/G1期阻滞,细胞凋亡程度的增加及伴随胞内Caspase-3活性的增加。各实验组与对照组比较,差异均有统计学意义(P0.01)。Western blot法结果显示随着药物浓度的增加,抗凋亡蛋白Bcl-2下调、促凋亡蛋白Bax表达明显上调,各实验组与对照组相比,差异有统计学意义(P0.01)。结论硫利达嗪对乳腺癌细胞MDA-MB-231、MCF-7具有显著的增殖抑制作用,且可显著诱导肿瘤细胞发生G0/G1期阻滞及凋亡,伴随Caspase-3活性的上调,其毒性机制可能与下调肿瘤细胞内抗凋亡蛋白Bcl-2、上调Bax有关。
[Abstract]:Background & objective Breast cancer is one of the most common malignant tumors in women. The incidence of breast cancer increases year by year and tends to be younger. Especially, the malignant degree and prognosis of three-negative breast cancer with negative expression of ERPR and Her-2 are higher and worse. How to effectively study new drugs for Sanyin breast cancer has become the focus of breast cancer treatment [1-2]. Thirizine is a phenothiazine derivative, which is widely used in the treatment of severe mental and emotional disorders [3]. The latest study [4] confirmed that tiridamine could antagonize the activity of many kinds of tumor cells in vitro, suggesting that it may be used as a new anticancer drug for clinical treatment of cancer in the future. In this study, tri-negative breast cancer MDA-MB-231 cells and breast cancer MCF-7 cells were treated with tiridamine. The apoptosis of MDA-MB-231 and MCF-7 cells was observed and its mechanism was discussed. Methods the inhibitory effect of thiridamine on the cells was determined by MTT method and the half inhibitory concentration (IC50) was calculated. The effect of thiridamine on cell cycle distribution and apoptosis was detected by flow cytometry. Effect of drugs on the activity of Caspase-3 by colorimetric assay the expression of apoptosis-regulating protein Bcl-2nBax was detected by Western blot assay. Results the proliferation of MDA-MB-231 MCF-7 cells was inhibited in a dose-dependent manner after 24 h treatment with IC50 of 1822 渭 mol / L, respectively. The results of flow cytometry showed that with the increase of the concentration of thiridamine, MDA-MB-231 MCF-7 cells had different degrees of G0/G1 arrest, the degree of apoptosis and the increase of intracellular Caspase-3 activity. Compared with the control group, there were significant differences between the experimental group and the control group. The results of Western blot showed that with the increase of drug concentration, the anti-apoptotic protein Bcl-2 was down-regulated, and the expression of pro-apoptotic protein Bax was up-regulated, compared with the control group. The difference was statistically significant (P 0.01). Conclusion tiridazide can inhibit the proliferation of breast cancer cell line MDA-MB-231 and MCF-7, and induce G0/G1 phase arrest and apoptosis, accompanied by up-regulation of Caspase-3 activity. The mechanism of its toxicity may be related to the down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of Bax in tumor cells.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R737.9

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