MTBD抗肿瘤细胞活性及作用机制研究

Anti-tumor activity and molecular mechanisms of MTBD

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 detect the anti-tumor activity and molecular mechanisms of 4-[-2-（3-Methyl-thiophen-2- yl）-thiazol-4-yl]-benzene-l,2-dio（MTBD） as inhibitor of Aurora-B kinase. METHODS： The anti-tumor activity was de- tected by MTT assay. ELISA assay was performed to test the inhibition o{ purified recombinant human Aurora-B kinase. Flow cytometry analysis was used to analysis the cell cycle treated with MTBD. The relative mRNA expression level of Hela cells treated with MTBD was detected by real time PCR. RESULTS： MTBD could inhibit the growth of tumor cells, such as Hela,HepG2 and A549,and its ICs0 was （1. 034±2.23）,（1.72±3.78） and （2.01±1.23）μmol/L respectively; MTBD could induce S phase and Gz/M phase arrest and apoptosis in Hela cells. MTBD could also inhibited the activity of Aurora-B kinase and its IC50 was （1.70±2.02）μmol/L. But the H3 （Serl0） phosphorylation increased treated with MT- BD. RT-PCR analysis showed that MTBD could increase the mRNA expression level of p21 and p53 and decrease the lev- el of CDK2,Cyclin A1 and pCNA. CONCLUSIONS：MTBD can inhibit the proliferation of tumor cells. MTBD can induce cell cycle arrest and apopsis through several cell cycle events but not Aurora-B kinase.