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目的:对新发现的Aurora-B激酶抑制剂4-[2-(3-甲基噻吩-2-基)噻唑-4-基]-苯-1,2-二醇(MTBD)进行肿瘤细胞增殖抑制活性和作用机制的研究。方法:MTT法检测MTBD对肿瘤细胞的增殖抑制活性;流式细胞术检测MTBD对Hela细胞的周期阻滞和组蛋白3的磷酸化;ELISA方法检测MTBD对Aurora-B激酶的抑制活性;RT-PCR技术检测MTBD对S期检查点的相关蛋白转录水平的影响。结果:MTBD能够抑制Hela细胞、HepG2细胞和A549细胞的增殖,其IC50分别为(1.03±2.23)、(1.72±3.78)和(2.01±1.23)μmol/L;MTBD能够诱导Hela细胞发生G2/M期和S期阻滞,并诱导细胞发生凋亡;MTBD能够抑制Aurora-B激酶活性,其IC50为(1.70±2.02)μmol/L,但未能检测到Hela细胞内组蛋白3(Ser10)磷酸化水平下降;S期检查点相关基因p21和p53转录上调,CDK2、Cyclin A1和pCNA转录不同程度下调。结论:MTBD能抑制肿瘤细胞增殖,诱导细胞周期阻滞和凋亡;MTBD抗肿瘤活性的发挥可能不是主要通过Aurora-B激酶抑制,而是与多个细胞周期因子相关,并且对于不同肿瘤细胞株有不同的作用机制。
OBJECTIVE: To perform tumor cell proliferation of the newly discovered Aurora-B kinase inhibitor 4- [2- (3-methylthiophen-2-yl) thiazol-4-yl] -benzene-1,2-diol (MTBD) Inhibition of activity and mechanism of action. Methods MTT method was used to detect the proliferation inhibitory activity of MTBD on tumor cells. Cytotoxicity of MTBD on Hela cells and phosphorylation of histone 3 were detected by flow cytometry. The inhibitory activity of MTBD on Aurora-B kinase was detected by ELISA. PCR technique was used to detect the effect of MTBD on the transcription level of related proteins at S phase checkpoint. Results MTBD could inhibit the proliferation of Hela cells, HepG2 cells and A549 cells with the IC50 of (1.03 ± 2.23), (1.72 ± 3.78) and (2.01 ± 1.23) μmol / L, respectively. MTBD could induce the proliferation of Hela cells in G2 / M (1.70 ± 2.02) μmol / L, MTT could inhibit Aurora-B kinase activity, but failed to detect Hela intracellular histone 3 (Ser10) phosphorylation The levels of p21 and p53 were up-regulated at the checkpoint of S phase and the transcription of CDK2, Cyclin A1 and pCNA was down-regulated to some extent. CONCLUSION: MTBD can inhibit tumor cell proliferation, induce cell cycle arrest and apoptosis. The anti-tumor activity of MTBD may not be mainly inhibited by Aurora-B kinase, but related to many cell cycle factors, and for different tumor cell lines There are different mechanisms of action.