MDM2/MDMX-p53双靶点蛋白增效厄洛替尼抗胰腺癌活性的研究

Anti-Cancer Activity of Erlotinib Combined with MDM2/MDMX-p53 Inhibitor in Pancreatic Cancer Cells

目的:因对总生存时间的提高仅0.4个月,厄洛替尼在胰腺癌中的应用一直存在争议。本课题组前期成功合成了可阻断MDM2/MDMX-P53的抑制蛋白,本研究拟通过联合该抑制蛋白探讨提高厄洛替尼在胰腺癌中的抗肿瘤活性的可行性。方法:MTT法检测细胞增殖、流式细胞仪测定细胞凋亡、Matrigel transwell检测细胞侵袭能力,了解联合双靶点抑制蛋白对比厄洛替尼单药对细胞生物活性的影响,Western-blotting方法检测相关蛋白表达情况探讨联合用药的抗肿瘤增效机制。构建裸鼠移植瘤模型研究双靶点抑制蛋白联合厄洛替尼的体内抗肿瘤活性。结果:联合双靶点抑制蛋白可上调P53表达,下调MDM2、MDMX和EGFR表达,从而促进厄洛替尼抑制胰腺癌细胞增殖、促进凋亡、抑制细胞侵袭转移;体内研究显示双靶点抑制蛋白有协同抗肿瘤作用,各组移植瘤抑瘤率分别为:厄洛替尼22.47%、MDM2/MDMX-p53抑制蛋白组28.41%、联合治疗组55.39%,差异有统计学意义(P<0.01)。结论:联合MDM2/MDMX-P53双靶点抑制蛋白可增加厄洛替尼的体内、体外抗胰腺癌活性,有望为胰腺癌的治疗带来新的希望。

Objective:The clinical application of erlotinib in patients with pancreatic cancer has been controversial due to its limitation in improving overall survival(only 0.4 months).Our study aimed to enhance the anti-pancreatic cancer activity of erlotinib by combing the MDM2/MDMX-p53 inhibitor.Methods:For the in vitro study,MTT was applied to detect cell viability;cell apoptosis was evaluated by flow cytometry;Matrigel transwell assays were employed to detect cell invasion ability.Western blotting was used to learn the expression level of associated proteins in order to reveal the possible mechanisms.Nude mice subcutaneous transplantation tumor models were established to learn the in vivo anti-tumor activity of erlotinib combined with MDM2/MDMX-p53 inhibitor.Results:For the in vitro study,MDM2/MDMX-p53 inhibitor could significantly inhibit cell proliferation,promote cell apoptosis,and impair cell invasion.For the in vivo study,the inhibition rates were 22.47%(the erlotinib group),28.41%(the MDM2/MDMX-p53 inhibitor group),55.39%(the combination group),respectively;and the difference was statistically significant(P<0.01).Further study revealed the combined treatment showed MDM2/mdmx-p53 inhibitor could up-regulate p53,downregulate MDM2,MDMX and EGFR,which might contribute to the improvement of the anti-cancer activity of erlotinib.Conclusion:MDM2/MDMX-p53 inhibitor can improve the anti-tumor activity of erlotinib in pancreatic cancer both in vitro and in vivo.Further studies were needed if the combined treatment could be developed as a promising strategy for pancreatic cancer in clinic.