2-羟基-3-甲基蒽醌诱导结肠癌细胞凋亡机制的研究

Mechanism of 2-hydroxy-3-methylanthraquinone on inducing the apoptosis of colon cancer cells

目的探讨2-羟基-3-甲基蒽醌(HMA)对结肠癌HCT116和HT-29细胞生长的抑制的影响及其可能的机制。方法通过TCMSP数据库检索分析HMA的作用靶点。体外培养结肠癌细胞HCT116和HT-29,并给予不同浓度的HMA干预,通过CCK-8、台盼蓝计数、集落形成和Annexin V/PI双染法检测HMA对结肠癌细胞活力、生长、存活能力及凋亡的影响,Western blot法检测HMA对潜在靶点及凋亡调节蛋白表达的影响。结果TCMSP检索分析得到HMA存在于白花蛇舌草、半枝莲和巴戟天中药,含有32个潜在靶点,其中包含凋亡相关基因Caspase-3。不同浓度的HMA干预结肠癌细胞可抑制细胞活力、细胞生长、集落形成数量,同时诱导细胞凋亡。Western blot结果显示,HMA可下调抗凋亡基因Bcl-2,上调促凋亡基因Bax,促进凋亡相关蛋白cleaved Caspase-9和cleaved Caspase-3蛋白的活化、剪切,从而促进细胞凋亡。结论HMA对大肠癌细胞有抑制生长和诱导凋亡的作用,可能是通过调控Bcl-2、Bax和cleaved Caspase-9和cleaved Caspase-3等相关凋亡蛋白的表达。

Objective To explore the effects and possible mechanism of 2-hydroxy-3-methylanthraquinone(HMA)on the growth inhibition of colon cancer cells(HCT116,HT-29).Methods The target of HMA was searched and analyzed by TCMSP database.The colon cancer HCT116 and HT-29 cells were cultured in vitro,and then they were intervened with different concentrations of HMA.The effects of HMA on the viability,growth,survival ability and apoptosis of colon cancer cells were detected by CCK-8,trypan blue count,colony formation and Annexin V/PI double staining respectively.The effects of HMA on potential targets and apoptosis-regulating proteins were detected by Western blot.Results TCMSP search analysis showed that there was HMA in Hedyotis diffusa,Scutellaria barbata and Morinda officinalis,which contained 32 potential targets,including apoptosis-related gene Caspase-3.Different concentrations of HMA could inhibit cells viability,growth and number of formed colonies,and induce apoptosis.The results of Western blot showed that HMA could down-regulate anti-apoptosis gene Bcl-2,up-regulate pro-apoptosis gene Bax,and promote the activation and cleavage of apoptosis-related proteins(cleaved Caspase-9,cleaved Caspase-3),thus promoting apoptosis.Conclusion HMA can inhibit growth and induce apoptosis of colon cancer cells,which may be related to regulating expressions of Bcl-2,Bax,cleaved Caspase-9,cleaved Caspase-3 and other apoptosis proteins.