细胞周期蛋白质依赖激酶抑制药抗胰腺癌细胞增殖活性的研究

Anti-proliferation activity study of cyclin-dependent kinase inhibitors in pancreatic cancer cells

目的 探索细胞周期蛋白依赖性激酶(CDK)抑制药治疗胰腺癌的应用潜力。方法 用噻唑蓝法测定了16个CDK抑制药对人胰腺癌MIA PaCa-2细胞的增殖抑制活性,并通过流式分析和蛋白质印迹实验考察了高活性CDK抑制药的抗肿瘤机制。结果 在所有受试CDK抑制药中,CDK2/7/9抑制药SNS-032在给药48 h后对MIA PaCa-2细胞增殖的抑制活性最强[半抑制浓度(IC_(50))为(4.44±0.44)μmol·L^(-1)]。流式细胞分析显示:SNS-032引起MIA PaCa-2细胞周期轻度阻滞在S期,并能够呈浓度依赖性地诱导细胞凋亡。蛋白质印迹实验结果显示:SNS-032能浓度依赖性地下调MIA PaCa-2细胞中磷酸化视网膜母细胞瘤(p-Rb)、原癌基因(c-Myc)和髓样细胞白血病-1(Mcl-1)蛋白表达。在给药72 h后,SNS-032对携带不同kras基因型的多株人胰腺癌细胞的抑制活性(IC_(50)为0.18~0.91μmol·L^(-1))均强于阳性药吉西他滨(IC_(50)为0.32~3.92μmol·L^(-1))。结论 CDK2/7/9抑制药SNS-032能够抑制胰腺癌细胞增殖,并通过抑制CDK工作网络,诱导MIA PaCa-2细胞凋亡。

Objective To explore the development potential of cyclin-dependent kinase(CDK) inhibitor for treatment of pancreatic cancer.Methods The anti-proliferative activity of 16 CDK inhibitors were screened in human pancreatic cancer MIA PaCa-2 cells in methyl thiazolyl tetrazolium assay.The antitumor mechanism of highly active CDK inhibitor was investigated by both flow cytometry analysis and Western blot.Results SNS-032,a CDK2/7/9 inhibitor,exhibited the most potent in vitro ant-proliferative activity [50% inhibiting concentration(IC_(50)) was(4.44±0.44) μmol·L^(-1)] among all test compounds after 48 h exposure in MIA PaCa-2 cells.The flow cytometry analysis indicated that SNS-032 could slightly induce cell cycle arrested in S phase,and a dose-dependent apoptosis was observed in MIA PaCa-2 cells.In addition,Western blot indicated that SNS-032 dose-dependently down-regulated the protein expression of phospho-retinoblastoma(p-Rb),oncogene(c-Myc) and myeloid cell leukemia-1(Mcl-1) in MIA PaCa-2 cells.SNS-032 exhibited higher potency(IC_(50) was 0.18-0.91 μmol·L^(-1)) than positive control drug gemcitabine(IC_(50 )was 0.32-3.92 μmol·L^(-1)) on cell proliferation in multiple pancreatic cancer cells with different geotypic kras after 72 h exposure.Conclusion CDK2/7/9inhibitor SNS-032 can inhibit the proliferation of pancreatic cancer cells and induce apoptosis of MIA PaCa-2 cells by inhibiting the CDK/network.