以二氢吡唑为先导的新型化合物靶向hTERT抑制SMMC-7721的增殖

Novel compound containing dihydropyrazole moiety inhibits the proliferation of SMMC-7721 by targeting hTERT

目的观察以二氢吡唑为先导的新型化合物对人肝癌细胞株SMMC-7721增殖抑制的影响。方法采用噻唑蓝(MTT)比色法筛选对SMMC-7721细胞株增殖的抑制作用较强的小分子化合物,并用流式细胞周期实验观察化合物对细胞周期的影响,采用改良的端粒重复序列扩增程序(TRAP)实验检测端粒酶的活性,Western blot法检测化合物对h TERT蛋白表达的影响。结果与香豆素类衍生物相对比,新型二氢吡唑类化合物显示出较好的抑制肿瘤细胞增殖的作用;流式细胞周期检测结果显示该化合物通过S期阻滞影响细胞增殖;改良的TRAP实验结果显示该化合物对端粒酶的活性具有较明显的抑制作用;Western blot法结果显示该化合物可以显著降低h TERT蛋白的表达。结论以二氢吡唑为先导的新型化合物具有抑制SMMC-7721细胞的增殖的作用和周期阻滞作用,初步发现这主要是通过靶向h TERT抑制端粒酶的活性实现的。

Objective To investigate the inhibitory effect of novel compound containing dihydropyrazole moiety on the proliferation of SMMC-7721 by targeting hTERT melittin. Methods The inhibition rate of proliferation of cells treated with novel compounds was measured by MTT assay. And the novel compounds’ effect on cell cycle was test-ed by flow cytometry cell cycle experiment. Telomerase activity was determined through modified Telomeric Repeat Amplification Protocol （TRAP） assays. The protein expression level of hTERT was observed by Western blot. Re-sults Compared with the heterocyclic compounds, novel compound containing dihydropyrazole moiety had obvious inhibitory effect on the proliferation of cell lines, especially on the human hepatoma cell line SMMC-7721. The re-sult of flow cytometric cell cycle analysis showed that the number of cells in S phase was markedly increased. After the treatment with novel compound containing dihydropyrazole moiety, the results of modified TRAP assays predic-ted that the telomerase activity was inhibited. And the results of western blot showed that the protein expression level of hTERT decreased obviously. Conclusion Novel compound containing dihydropyrazole moiety inhibits the prolif-eration of SMMC-7721 and makes them arrested at S phase. This initial discovery is mainly achieved through the inhibition of telomerase activity by targeting hTERT.