多巴酚丁胺对顺铂作用下人肺腺癌细胞株A549生物学行为及YAP蛋白表达的影响

发布时间：2018-06-24 11:30

本文选题：多巴酚丁胺 + 顺铂　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:研究多巴酚丁胺单独或联合顺铂(DDP)对人肺腺癌细胞A549生物学行为的影响,探讨多巴酚丁胺和DDP联合运用治疗肺腺癌的可能性。方法:选取人肺腺癌细胞A549,并将细胞分成对照(control)组、多巴酚丁胺组、DDP组和联合组(多巴酚丁胺+DDP),进行如下实验:(1)CCK-8法:对加入多巴酚丁胺(0、7.5、15、30、60、120μmol/L)或DDP(0、1.25、2.5、5、10、20μg/ml)分别干预24、48、72h后A549细胞的增殖水平进行检测,并选择合适的作用浓度和时间;(2)划痕愈合实验:检测各组A549细胞迁移能力的变化;(3)Transwell侵袭实验:对各组A549细胞侵袭能力的变化进行检测;(4)流式细胞术:对各组细胞凋亡率的变化进行检测;(5)Western Blot技术:对各组细胞Yes相关蛋白(YAP)和磷酸化YAP(P-YAP)的表达水平进行检测。结果:(1)细胞增殖水平的检测:多巴酚丁胺和DDP都可以抑制A549细胞的增殖,细胞的增殖抑制率呈浓度和时间依赖性增加(P0.05)。(2)细胞迁移能力的检测:与control组相比,多巴酚丁胺组A549细胞的划痕愈合率降低(P0.05);与多巴酚丁胺或DDP单独作用组相比,联合组A549细胞的划痕愈合率降低(P0.05)。(3)细胞侵袭能力的检测:与control相比,多巴酚丁胺组的穿膜细胞数减少(P0.05);与多巴酚丁胺或DDP单独作用组相比,联合组的穿膜细胞数减少(P0.05)。(4)细胞凋亡率的检测:与control组相比,多巴酚丁胺组A549细胞的凋亡率增加(P0.05);与多巴酚丁胺或DDP单独作用组相比,联合组A549细胞的凋亡率增加(P0.05)。(5)YAP和P-YAP蛋白表达水平的检测:与control组相比,多巴酚丁胺组和顺铂组A549细胞YAP的表达都减少,并且P-YAP的表达都增加(P0.05);与多巴酚丁胺组相比,联合组A549细胞YAP的表达减少,并且P-YAP的表达增加(P0.05)。结论:多巴酚丁胺可能是通过抑制人肺腺癌A549细胞YAP蛋白的表达协同增强DDP的作用,进而抑制A549细胞的增殖、迁移和侵袭转移能力,并促进A549细胞的凋亡。
[Abstract]:Aim: to study the effect of dobutamine alone or in combination with cisplatin (DDP) on the biological behavior of human lung adenocarcinoma cell line A549 and to explore the possibility of dobutamine combined with DDP in the treatment of lung adenocarcinoma. Methods: human lung adenocarcinoma cell A549 was selected and divided into control (control) group. Dobutamine group and dobutamine group (dobutamine group) were tested as follows: (1) CCK-8 method: the proliferation level of A549 cells was detected by adding dobutamine (07.5N 15303060120 渭 mol / L) or DDP (0 1. 252.5A1020 渭 g/ml). And select the appropriate concentration and time; (2) scratch healing test: detect the change of migration ability of A549 cells in each group; (3) Transwell invasion test: to detect the change of invasion ability of A549 cells in each group; (4) flow cytometry: to detect the changes of cell migration ability of A549 cells in each group; (4) flow cytometry: to detect the migration ability of A549 cells in each group. (5) Western blot: the expression levels of YAP and phosphorylated YAP (P-YAP) were detected. Results: (1) the level of cell proliferation: dobutamine and DDP could inhibit the proliferation of A549 cells. The inhibition rate of cell proliferation was increased in a concentration and time dependent manner (P0.05). Compared with control group, dobutamine and DDP could inhibit the proliferation of A549 cells in a concentration-and time-dependent manner. Compared with dobutamine or dobutamine alone, the rate of scratch healing of A549 cells in dobutamine group was lower than that in dobutamine alone group (P0.05). (3). Compared with dobutamine or dobutamine alone, the number of perforated cells decreased (P0.05) in dobutamine group (P0.05). Compared with control group, the number of perforated cells in combination group decreased (P0.05 / 4). The apoptosis rate of A549 cells in dobutamine group was higher than that in dobutamine alone group (P0.05), and the expression level of Yap and P-YAP protein in combination group was higher than that in control group. In dobutamine group and cisplatin group, the expression of YAP in A549 cells decreased, and the expression of P-YAP increased (P0.05). Compared with dobutamine group, the expression of YAP in A549 cells decreased and the expression of P-YAP increased in combination group (P0.05). Conclusion: dobutamine may inhibit the expression of YAP protein in human lung adenocarcinoma cell line A549, and then inhibit the proliferation, migration, invasion and metastasis of A549 cells, and promote the apoptosis of A549 cells.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R734.2

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