重组人血管内皮抑素联合放疗对肺腺癌A549细胞TGF-β1、HIF-1α表达的影响

Effect of Recombinant Endostatin in Combination with Radiotherapy on the Expression of TGF-β1 and HIF-1α in Lung Adenocarcinoma Cell A549

摘要:

通过观察Endostar联合放疗对肺腺癌A549细胞增值及TGF-β1、HIF-1α表达的影响，初步探讨重组血管内皮抑素的放疗增敏机制。方法：将培养至对数生长期的A549细胞分为对照组、单纯放疗组、恩度组、联合组，检测各组A549细胞增殖率及TGF-β1、HIF-1α蛋白和mRNA表达情况。结果：1) 除对照组外，各组细胞的增殖随时间的改变都受到不同程度的抑制，以联合组最为明显，差异具有统计学意义(p < 0.05)；2) 处理后各组A549细胞TGF-β1、HIF-1α的分泌下降，24 h以放疗组及联合组TGF-β1、HIF-1α表达量明显下降 (p < 0.05)；3) PCR扩增后，各处理组 TGF-β1、HIF-1α mRNA的表达随着时间变化均表现出下降趋势，联合组在24 h差异有显著统计学意义(p < 0.05)。结论：恩度联合放疗可显著抑制肺腺癌A549细胞增殖及TGF-β1、HIF-1α的表达，恩度可能通过下调TGF-β1、HIF-1α的表达进而发挥放疗增敏作用。 To observe the effect of Endostar combined with Radiotherapy on the expression of Transforming Growth Factor-beta 1 (TGF-β1) and Hypoxia-induced factor-1 (HIF-1α) in lung adenocarcinoma A549 cells and to discuss the possible mechanisms. Methods: To culture the A549 cell, when the cells achieve Logarithmic phase and there are four groups: Control group; Endostar group; Radiotherapy group; Endostar in combination with Radiotherapy. Cell proliferate rate was measured by CCK8, and the expression level of TGF-beta 1 and HIF-1α protein and mRNA of A549 cells in each group were examined by ELISA and PCR. Result: 1) Besides control group, the proliferation of tumor cells in each group are subject to different degree of inhibition. Compared with control and Endostar groups, there is a significant difference of proliferate inhibition in Endostar combined with Radiotherapy group (p < 0.05). 2) After treatment, the expressions of TGF-β1 and HIF-1 of A549 cells in each group appears to be declining differently. The Radiotherapy group and combined group have a significant difference in cell proliferation at 24 hours, compared to control group (p < 0.05). 3) After PCR amplification, the changes of expressions of TGF-β1 and HIF-1α mRNA over time (0 - 24 hours) compared with the control group, all treatment groups showed a decreasing trend. After 24 hours of treatment, Compared with control group and Endostar group, the difference in the combined group was statistically sig- nificant (p < 0.05). Conclusion: Endostar combined with Radiotherapy could significantly enhance the inhibition of proliferate of cells and expressions of TGF-β1 and HIF-1α in tumor cells. Thus enhance the inhibition of tumor angio- genesis and metastatic.