Expression Establishment And Bionomics Analysis Of The Drug-Resisitance A549-ABCG2 Cell Line

OBJECTIVE:This study was to establish the functional expression of ABCG2 in lung adenocarcinoma cell line A549 and to observe the change of bionomics,provide an ideal experimental platform for reseaching the mechanism of ABCG2—mediated drug resistance,and developing effective metOD s to reverse the drug resistance.METHOD:After extract total RNA from the tissue of NSCLC patients by trizol,reverse transcriptase-polymerase chain reaction(RT-PCR)was performed and ABCG2-pcDNA3.1 plasmid was transferred into A549 cells.Stable expression of ABCG2 was established in A549 cells through G418 selection.And clones which named A549-ABCG2 were taken and cultivated in amplification.Positive ABCG2 cells which showed well expression were selected by both RT-PCR and Western blot.The function of ABCG2 was reseached by flow cytometry after adding doxorubicin or hydroxycamptothecin.A549 and A549-ABCG2 were tested with MTT assay to evaluate their chemoresistance to different anti-cancer drugs.The ATPase assay was also performed to reflect the activity of ABCG2.RESULT:The positive A549-ABCG2 cell show well expression of ABCG2 and ability of resistting the effect of anti-cancer drugs when detected by flow cytometry.The apoptotic population in A549 cells are more than in ABCG2 positive cells and A549 showed a apoptotic peak with 6.7%apoptosis rate after applying doxorubicin. A549-ABCG2 cells decreased 14.7%in S period and rised 13.5%in G1 period, while A549 cells rised 18.9%in S period and decreased 18.5%in G1 period with hydroxycamptothecin.The sensitivity of A549-ABCG2 cell to chemotherapy medicines was lower than A549 cell and the activity of ATPase was higher than the latter.CONCLUSION:We can use ABCG2-over-expressed A549-ABCG2 cell as a tool to select sensitive anti-cancer drugs in clinical treatment of ABCG2 high expressed patients.