钙调蛋白依赖性蛋白激酶Ⅱ在维拉帕米逆转肺腺癌顺铂化疗耐药中的作用

刘淼; 范平生; 张腾跃; 黄金; 樊高飞; 刘亚贝

doi:10.7619/jcmp.201910002

钙调蛋白依赖性蛋白激酶Ⅱ在维拉帕米逆转肺腺癌顺铂化疗耐药中的作用

Role of calmodulin-dependent protein kinase Ⅱ in reversingdrug resistance to cisplatin chemotherapy by verapamil in patients with lung adenocarcinoma cells

摘要

摘要:
目的探讨钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)在维拉帕米逆转肺腺癌顺铂化疗耐药中的作用。
方法在肺腺癌细胞系(A549)中，采用CCK-8法检测维拉帕米逆转顺铂耐药的能力，采用流式细胞仪Annexin V-FITC/PI法检测维拉帕米逆转顺铂耐药中凋亡水平的效果，采用Western blotting检测逆转耐药过程中P-CaMKⅡ/CaMKⅡ蛋白表达水平的变化。
结果在肺腺癌细胞系(A549)中，单药顺铂组、顺铂联合维拉帕米组的细胞增长抑制率分别为(55.00±2.60) %、(32.30±1.80) %, 差异有统计学意义(P < 0.05)。凋亡实验表明，单药顺铂组细胞凋亡率为10.30%, 顺铂联合维拉帕米组为21.60%, 差异有统计学意义(P < 0.05)。Western blotting结果显示，在A549细胞中，顺铂联合维拉帕米组P-CaMKⅡ/CaMKⅡ的表达明显低于单药顺铂组。
结论在肺腺癌细胞中, CaMKⅡ参与维拉帕米逆转化疗耐药的过程。

Abstract:
Objective To investigate the role of calmodulin-dependent protein kinase Ⅱ(CaMKⅡ) in reversing drug resistance to cisplatin chemotherapy by verapamil in patients with lung adenocarcinoma cells.
Methods In lung adenocarcinoma cell lines (A549), the CCK-8 method was used to detect the ability of verapamil in reversing the drug resistance to cisplatin chemotherapy, the Annexin V-FITC/PI method was used to detect the effect of verapamil on reversing the apoptosis of drug resistance to cisplatin chemotherapy, and the Western blotting was used to detect the expression levels of P-CaMKⅡ/CaMKⅡ protein in reversal process.
Results In the A549 cell line, the inhibition rates of DDP group and DDP-VER combination group were (55.00±2.60) % and (32.30±1.80) % respectively, and there was significant difference between the two groups (P < 0.05). The apoptosis rates of DDP group and DDP-VRP combination group were 10.30% and 21.60% respectively, and there was significant difference between the two groups (P < 0.05). Western blotting results showed that the expression level of P-CaMKⅡ/CaMKⅡ protein in A549 cell line in DDP-VRP combination group was obviously lower than that in DDP group.
Conclusion In lung adenocarcinoma cells, CaMKⅡ participates in the reversal process of drug resistance to cisplatin chemotherapy by verapamil.

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