Effect of the p53 alpha gene on the chemosensitivity of the H1299 human lung adenocarcinoma cell line

摘要

To investigate the effects of tumor protein p53 (p53 or TP53) alpha gene on the chemosensitivity of the H1299 human lung adenocarcinoma cell line, the recombinant vector pEGFP-p53 alpha was constructed. The vector pEGFP-p53 alpha was transfected into the cultured p53-null H1299 cells using Lipofectamine 2000. The G418-resistant cells were then selected. The expression of the p53 alpha gene in these cells was examined using reverse transcription-polymerase chain reaction, and TP53 protein expression was examined using western blot analysis and immunocytochemistry. An MTT assay and colony formation assay were used to analyze the response of the transfected cells to cisplatin (CDDP). DAPI staining was used to determine the level of apoptosis of the transfected cells. The transfected H1299 human lung adenocarcinoma cells stably expressed TP53 protein. The MTT assay demonstrated that the 50% inhibitory concentrations for the H1299, H1299/pEGFP-N1 and H1299/pEGFP-p53 alpha cells were 28, 24 and 18 mu mol/l, respectively. The survival rate of H1299/pEGFP-p53 alpha cells was significantly reduced compared with that of H1299 and H1299/pEGFP-N1 cells (P<0.05). The colony formation assay and DAPI staining identified that the colony formation rate and the number of apoptotic cells of H1299/pEGFP-p53 alpha were significantly reduced, compared with those of the H1299 and H1299/pEGFP-N1 cells (P<0.05). Therefor, the present study demonstrated that the transfection of H1299 cells with the p53 alpha gene resulted in an increase in sensitivity to CDDP chemotherapy. The combination of CDDP and gene therapy for H1299 lung adenocarcinoma cell line provides an experimental basis for clinical research.