环磷酰胺联合唑来膦酸的用药顺序对乳腺癌MDA-MB-231细胞杀伤的影响

发布时间：2018-05-30 09:44

本文选题：乳腺癌 + 唑来膦酸　；参考：《郑州大学》2017年硕士论文

【摘要】：乳腺癌作为女性最常见的恶性肿瘤之一,因具有高度异质性,现有的治疗难以达到理想的治疗效果,尤其是那些ER、HER-2与PR表型均为阴性的三阴性乳腺癌患者,更无有效的治疗方案。因此,如何开发出新型药物和新的用药方案以提高乳腺癌的治疗效果是肿瘤研究当前急需解决的问题。目的:唑来膦酸联合甲磺酸伊马替尼治疗白血病具有协同作用,同时联合内分泌药物可延长乳腺癌患者生存期,但其与化疗药物对乳腺癌的抗肿瘤效果至今尚无报道。因此,本课题拟开展环磷酰胺联合唑来膦酸的不同用药顺序对人三阴性乳腺癌MDA-MB-231细胞的杀伤效果的研究,一方面探讨这种联合用药顺序的最佳效果,另一方面分析两者联合用药的功能机制,为三阴性乳腺癌的治疗方案提供新的参考和实验基础,为开发新的肿瘤化疗用药方案提供理论依据。方法:我们将实验分为空白对照组、环磷酰胺单药组、唑来膦酸单药组、唑来膦酸和环磷酰胺同时用药组、环磷酰胺贯序唑来膦酸用药组和唑来膦酸贯序环磷酰胺组六种用药模式。然后,采用CCK-8试剂分别检测不同浓度唑来膦酸和环磷酰胺对人乳腺癌MDA-MB-321细胞杀伤效果,同时计算出适于联合用药浓度。再此基础上,采用CCK8试剂、Celcein-AM/PI凋亡检测试剂盒和transwell小室的方法系统分析唑来膦酸和环磷酰不同用药顺序对人乳腺癌MDA-MB-231细胞杀伤和抑制迁移的效果。结果:1.CCK8结果显示:与正常对照组相比,环磷酰胺或唑来膦酸单独用药对MDA-MB-231细胞都具有一定的生长抑制作用,而且具有剂量依赖性和时间依赖性(P0.05);同时环磷酰胺和唑来膦酸72h对MDA-MB-231细胞的IC50分别为1μmol/L和12.5μmol/L,由此说明,环磷酰胺抗乳腺癌效果的作用比唑来膦酸的作用强。2.不仅如此,分析环磷酰胺联合唑来膦酸的不同用药顺序组对MDA-MB-231细胞的杀伤作用的影响,即使是低浓度的0.1μmol/L环磷酰胺与3.125μmol/L唑来膦酸进行组合,CCK8结果、Celcein-AM/PI凋亡检测结果和Transwell迁移实验均证明:与其他实验组相比,环磷酰胺序贯唑来膦酸的协同作用最强,它不仅可以诱导大量乳腺癌MDA-MB-231细胞凋亡,还能显著抑制MDA-MB-231细胞的迁移能力(P0.05)。结论:唑来膦酸联合环磷酰胺具有协同作用,这种协同作用受不同用药顺序的影响,其中以环磷酰胺序贯唑来膦酸抗肿瘤效果最佳,不仅可以有效地诱导MDA-MB-231细胞凋亡,还能抑制MDA-MB-231细胞迁移,这结果暗示着此联合用药方式在杀伤肿瘤细胞的同时,还能预防肿瘤细胞转移的发生,提高转移性乳腺癌患者的治疗效果。
[Abstract]:Breast cancer is one of the most common malignant tumors in women. Due to its high heterogeneity, the current treatment is difficult to achieve ideal therapeutic effect, especially in those patients with negative phenotypes of ERHER-2 and PR. There is no effective treatment. Therefore, how to develop new drugs and new regimen to improve the therapeutic effect of breast cancer is an urgent problem in cancer research. Objective: Zoledronic acid combined with imatinib mesylate has synergistic effect in the treatment of leukemia, and combined with endocrine drugs can prolong the survival time of breast cancer patients. However, the antitumor effect of zoledronic acid combined with chemotherapeutic drugs on breast cancer has not been reported. Therefore, this paper intends to study the killing effect of cyclophosphamide combined with zoledronic acid on human three-negative breast cancer MDA-MB-231 cells. On the other hand, the functional mechanism of combined use of the two drugs is analyzed, which provides a new reference and experimental basis for the treatment of triple negative breast cancer, and provides a theoretical basis for the development of a new chemotherapy regimen. Methods: we divided the experiment into blank control group, cyclophosphamide single drug group, zoledronic acid and cyclophosphamide group. Cyclophosphamide sequential zoledronic acid group and zoledronic acid sequential cyclophosphamide group six drug patterns. Then, CCK-8 reagent was used to detect the killing effect of different concentrations of zoledronic acid and cyclophosphamide on human breast cancer MDA-MB-321 cells, and to calculate the suitable concentration of combined use of zoledronic acid and cyclophosphamide. On the basis of this, the effects of zoledronic acid and cyclophosphoryl on killing and inhibiting migration of human breast cancer MDA-MB-231 cells were systematically analyzed by CCK8 reagent Celcein-AMPI assay kit and transwell chamber. Results 1. The results of CCK8 showed that cyclophosphamide or zoledronic acid alone could inhibit the growth of MDA-MB-231 cells. The IC50 of cyclophosphamide and zoledronic acid on MDA-MB-231 cells for 72 hours was 1 渭 mol/L and 12.5 渭 mol / L, respectively, which indicated that cyclophosphamide had stronger effect on breast cancer than zoledronic acid. Moreover, the effects of cyclophosphamide combined with zoledronic acid on MDA-MB-231 cells were analyzed. Even the low concentration of 0.1 渭 mol/L cyclophosphamide combined with 3.125 渭 mol/L zoledronic acid showed that the synergistic effect of cyclophosphamide sequential zoledronic acid was the strongest compared with other experimental groups. It can not only induce the apoptosis of breast cancer MDA-MB-231 cells, but also inhibit the migration ability of MDA-MB-231 cells. Conclusion: Zoledronic acid combined with cyclophosphamide has synergistic effect, and this synergism is affected by different drug sequence. Cyclophosphamide sequential zoledronic acid has the best antitumor effect, which can not only effectively induce apoptosis of MDA-MB-231 cells. The results suggested that the combined therapy could not only kill tumor cells, but also prevent the metastasis of tumor cells and improve the therapeutic effect of metastatic breast cancer patients.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9

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