多巴酚丁胺对顺铂作用下人肺癌细胞株A549生物学行为及YAP蛋白表达的影响

Effect of dobutamine on biological behavior and expression of YAP protein of human lung cancer cell line A549 treated with cis-diamminedichloroplatinum

目的 研究多巴酚丁胺对顺铂（DDP）作用下人肺腺癌细胞株A549生物学行为及YAP和磷酸化YAP（P-YAP）蛋白表达的影响.方法 体外培养人肺腺癌细胞株A549,实验分为对照组、多巴酚丁胺组、DDP组和多巴酚丁胺与DDP联合组.用CCK-8法检测各组细胞的增殖抑制率,Transwell法和划痕愈合实验检测各组细胞的体外侵袭能力和迁移能力,流式细胞术检测各组细胞凋亡率,Western blot法检测各组细胞YAP和P-YAP蛋白的表达.结果 多巴酚丁胺或DDP能够抑制A549细胞增殖,呈浓度和时间依赖性（P〈0.05）.对照组、多巴酚丁胺组、DDP组及多巴酚丁胺与DDP联合组的划痕愈合率分别为（41.22±0.70）%、（16.11±0.39）%、（14.49±0.25）%、（7.24±0.30）%,差异有统计学意义（P〈0.05）;穿膜细胞数分别为（262.91±2.81）、（115.03±4.26）、（81.88±3.91）、（48.51±3.99）个,差异有统计学意义（P〈0.05）;凋亡率分别为（22.28±0.61）%、（27.92±1.36）%、（32.16±1.80）%、（36.70±1.87）%,差异有统计学意义（P〈0.05）.与多巴酚丁胺组相比,多巴酚丁胺与DDP联合作用能够抑制YAP的表达及促进P-YAP的表达（P〈0.05）.结论 多巴酚丁胺可能通过抑制YAP的表达来增强DDP的作用,进而抑制人肺腺癌A549细胞增殖、体外侵袭能力,并促进细胞凋亡.

Objective To investigate the effect of dobutamine on biological behavior and the expressions of YAP and phospho-YAP （P-YAP） of human lung adenocarcinoma A549 cells treated with cis-diamminedichloroplatinum （DDP）. Methods Human adenocarcinoma A549 cells were cultured and divided into control group, dobutamine group, DDP group and dobutamine＋DDP group. The proliferation inhibitory rates of these groups were detected by CCK-8 assay. The invasive abilities of these groups were detected by Transwell method. The scratch healing experiment was used to test the migration and movement capacity of the cells. The apoptosis rates of these groups were analyzed by flow cytometry. The protein expressions of YAP and P-YAP of these groups were determined by Western blot. Results Dobutamine or DDP could inhibit the proliferation of A549 cells in a dose-dependent and time-dependent way （P 〈 0.05）. In control group, dobutamine group, DDP group and dobutamine ＋ DDP group, the scratch healing rates were （41.22 ±0.70） %, （16.11 ±0.39） %, （14.49 ±0.25） % and （7.24 ±0.30） %, respectively （P〈 0.05）, the transmembrane cell numbers were （262.91±2.81）, （115.03±4.26）, （81.88±3.91） and （48.51±3.99）, respectively （P〈 0.05）, and the apoptosis rates were （22.28±0.61）%, （27.92±1.36）%, （32.16±1.80）%and （36.70±1.87）%, respectively （P〈0.05）. The combination of dobutamine and DDP could inhibit the expression of YAP and promote the expression of P-YAP compared with dobutamine group （P〈0.05）. Conclusion Dobutamine may enhance the DDP effect by descending the expression of YAP to restrain the proliferation, invasion and migration, and to enhance the apoptosis of A549 cells.