Twist1基因在A549与A549/DDP细胞中的表达及意义

The Expression of Twist1 Gene in A549 and A549 / DDP Cells and its Significance

目的检测人Twist1基因在A549与A549/DDP细胞株中的表达,探讨Twist1与肺腺癌顺铂耐药的相关性。方法通过细胞计数,观察不同浓度梯度顺铂干预24 h后A549细胞和A549/DDP细胞的生长情况;用RIPA裂解液裂解A549、A549/DDP细胞,充分游离细胞总蛋白,用BCA法检测总蛋白浓度,ELISA法检测Twist1转录因子的浓度。结果定义m值(m=Twist1转录因子浓度/总蛋白浓度)。浓度为0、10、20、30、40、50 mg·L^(-1)的顺铂干预下,分别测得A549与A549/DDP的m值为3.01/3.31、2.17/2.82、1.63/2.94、1.73/1.99、0.37/1.31、-/-,A549/DDP细胞的m值显著大于A549细胞(P<0.05)。结论 Twist 1基因在人顺铂耐药肺腺癌细胞中表达增高,提示Twist1可能与肺腺癌顺铂耐药相关。

Objective To investigate the relationship between Twist1 and cisplatin resistance in lung adenocarcinoma by detecting the expression of Twist1 gene in A549 cell line and A549/DDP cell line. Methods The growth of A549 cells and A549/DDP cells was observed dynamically before and 24 h after the intervention of cisplatin with different concentration gradients, and cells were counted at the same time. The A549 cells and A549/DDP cells were split by RIPA Lysis Buffer to release the cell total protein completely. The total protein concentration was detected by enhanced BCA Protein Assay Kit, and the concentration of Twist1 transcription factor was detected by ELISA assay. Results The m＇ was defined as m = concentration of Twist1 transcription factor / concentration of total protein. The m of A549 cells and A549/DDP cells was 3.01/3.31, 2.17/2.82, 1.63/2.94, 1.73/1.99, 0.37/1.31,-/- respectively when intervened by 0, 10, 20, 30, 40, 50 mg·L-1 of cisplatin. The m of A549/DDP cells was significantly higher than that of A549 cells（P0.05）. Conclusion Twist 1 gene was highly expressed in human cisplatin-resistant lung adenocarcinoma cells. It may suggest a relationship between Twist1 and cisplatin resistance in lung adenocarcinoma.