Study on the reversion effect of targeting silence CXCR4 gene on Gemcitabine-resistance in non-small cell lung cancer / 中国生化药物杂志

ABSTRACT

Objective To explore the reversion effect of Gemcitabine-resistance A549 cell (A549/Gem) by silencing CXCR4.Methods A549 cell was induced by continuous stepwise exposure to Gemcitabine in order to obtain Gemcitabine-resistance A549 cell ( A549/Gem) in vitro.The CXCR4 expressions level of A549 and A549/Gem were detected by Quantitative RT-PCR ( RT-qPCR) and Western blot analyses.The CXCR4 shRNA vector was transfected into the A549/Gem cell by targeting silence CXCR4.Furthermore, MTT assay was used to explore the IC50 and RI in A549, A549/Gem and A549/Gem-CXCR4 cells.Moreover, Western blot analysis was performed to detect the expressions of phospho-JNK, phospho-p38 and phospho-ERK 1/2 in A549, A549/Gem and A549/Gem-CXCR4 cells.Results Gemcitabine-resistance A549 cell ( A549/Gem) was successful constructed by using continuous stepwise exposure to Gemcitabine in vitro.The expression level of CXCR4 was up-regulated in A549/Gem cell than in A549 cell.The CXCR4 shRNA vector could significantly decrease CXCR4 expression in A549/Gem cell.The IC50 values of Gemcitabine in A549, A549/Gem and A549/Gem-CXCR4 cell were (0.08 ±0.01)μmol/L, (14.01 ±0.21)μmol/L and (1.84 ±0.61)μmol/L, respectively.The RI value of Gemcitabine was (127.12 ±12.28) in A549/Gem cells, while the value of RI was (27.3 ±0.98) in A549/Gem-CXCR4 cells.The expression level of phospho-JNK, phospho-p38 and phospho-ERK 1/2 were also markedly inhibited in A549/Gem-CXCR4 cell than in A549/Gem cell.Conclusion CXCR4 is up-regulated in A549/Gem cell.Targeting silence CXCR4 can successfully reverse drug-resistance of Gemcitabine in A549/Gem cells, which hints CXCR4 is associated with lung cancer radiation therapy as an effective molecular target.