抑制Src酪氨酸激酶活性对人肺癌A549/DDP细胞耐药性及MDR1和LRP表达的影响  

Effect of Src Tyrosine Kinase Inhibition on the Drug-resistance as well as MDR1 and LRP Expression of the Human Cis-platinum-resistant Lung Cancer Cell Line A549/DDP

作　　者：律洁[1] 田玉峰[1] 

机构地区：[1]山东省日照市人民医院检验科,日照276826

出　　处：《中国肺癌杂志》2012年第9期501-506,共6页Chinese Journal of Lung Cancer

摘　　要：背景与目的本研究旨在探讨抑制Src酪氨酸激酶活性对人肺癌A549/DDP细胞耐药性及多药耐药蛋白(muti-drug resistance1,MDR1)和肺耐药相关蛋白(lungresistance-related protein,LRP)表达的影响。方法以Src酪氨酸激酶抑制剂作用于A549/DDP细胞,应用Western blot法检测肿瘤细胞Src酪氨酸激活性的变化,CellTiter-Glo发光法检测肿瘤细胞药物敏感性的变化,流式细胞仪检测肿瘤细胞Rh-123含量变化,Western blot法和RT-PCR检测肿瘤细胞MDR1和LRP表达变化。结果 Src酪氨酸激酶抑制剂可下调A549/DDP细胞中Src酪氨酸激活性,2.5M和10MSrc酪氨酸激酶抑制剂作用肿瘤细胞后,肿瘤细胞药物敏感性提高,逆转倍数(reversal fold,RF)分别为1.59倍和2.10倍,肿瘤细胞中Rh-123含量分别提高了1.21倍和1.59倍,MDR1mRNA表达分别是对照组的53.8%和27.5%,LRPmRNA表达分别是对照组的59.3%和21.4%,MDR1和LRP蛋白表达水平明显下降。结论抑制A549/DDP细胞中Src酪氨酸激酶活性可逆转肿瘤细胞多药耐药性,提高肿瘤细胞药物敏感性,其机制可能与降低细胞MDR1和LRP表达有关。Background and objective The aim of this study is to investigate the effect of Src tyrosine kinase inhibition on the drug-resistance as well as the expression of multidrug resistance 1 （MDR1） and lung resistance-related protein （LKP） of the human cis-platinum-resistant lung cancer cell line A549/DDP. Methods 4-Anilinoquirazoline was used to inhibit Src tyrosine kinase activity in AS49/DDP. Western blot analysis was used to detect the Src tyrosine kinase activity. CellTiter-Glo assay was used to detect the drug sensitivity of tumor cells. Flow cytometry was used to detect the intracellular Rh-123 content. Western blot and real-time PCR assay were used to detect the expression of tumor MDR1 and LRP. Results 4-Anilinoquirazoline can clown-regulate the cellular Src tyrosine kinase activity in A549/DDP. After treat- ment with 2.5 uM and 10 uM of 4-anilinoquirazoline, the cells became more sensitive to the drug and the reversal folds （RFs） of tumor cell sensitivity to the drug were 1.59- and 2.10-fold, respectively. The intracellular content of Rh-123 improved by 1.21- and 1.S9-fold, respectively. The mtLNA levels of MDgl were 53.8% and 27.5% of the control, respectively. The mRNA level of LKP was 59.3% and 21.4% of the control, respectively. The expression of MDR1 and LRP protein significantly de- creased. Conclusion The inhibition of Src tyrosine kinase activity in AS49/DDP cells can reverse multi-drug resistance and increase the sensitivity of the cells to the drug. The mechanism may be related to the down-regulation of cellular MDR1 and LRP.

关 键 词：SRC酪氨酸激酶 A549/DDP 多药耐药 MDR1 LRP 

分 类 号：R734.2[医药卫生—肿瘤]