摘要
目的研究血根碱与KRAS启动子G-四链体DNA的相互作用,探究以KRAS G-四链体DNA为靶点的血根碱对人结肠癌细胞SW620的作用及机制。方法采用MTT法检测血根碱对人结肠癌细胞SW620增殖的影响。采用紫外-可见吸收光谱、荧光光谱、圆二色谱等方法研究血根碱与KRAS启动子G-四链体DNA的相互作用。采用实时荧光定量PCR(Real-Time PCR)检测血根碱对SW620细胞KRAS mRNA表达的影响。结果血根碱能够抑制人结肠癌SW620细胞的增殖。血根碱与KRAS启动子G-四链体DNA的结合能力较强,结合计量比接近1∶1,结合分子机制为生成了不发光的复合物。血根碱可维持G-四链体DNA的平行构型及热稳定性。血根碱能抑制人结肠癌SW620细胞KRAS mRNA的表达。结论血根碱可能通过与KRAS启动子区域G-四链体结合并维持其结构的稳定,抑制KRAS mRNA表达,进而抑制人结肠癌细胞SW620的增殖,发挥抗癌作用。
Objective To explore the function and mechanism of sanguinarine on human colorectal cancer cells SW620 based on KRA S promoter G-quadruplex DNA target by investigating the interaction between sanguinarine and KRA S promoter G-quadruplex DNA. Methods The effects of sanguinarine on the proliferation of human colorectal cancer cells SW620 were detected by using MTT method. The interaction of sanguinarine and KRA S G-quadruplex DNA was investigated by methods of UV-Vis, fluorescence, and circular dichroism spectra. In addition, the effects of sanguinarine on the expression of KRA S mRNA of SW620 cells were detected by Real-time fluorescent quantitative PCR （Real-Time PCR）. Results Sanguinarine could inhibit the proliferation of human colorectal cancer cells SW620. Sangui- narine had a strong binding capacity with KRA S G-quadruplex DNA, and the stoichiometric ratio was about 1 ： 1. The binding molecular mechanism was the formation of nonluminous complex. Sanguinarine could keep the parallel structure of KRA S G-quadruplex DNA and its thermostability. It also could suppress the expression of KRA S mRNA of human colorectal cancer cells SW620. Condusion Sanguinarine may suppress the expression of KRAS mRNA, and further inhibit the proliferation of human colorectal cancer cells SW620 by combining with G-quadruplex in the KRAS promoter region and stabilizing its thermostability, so as to play the antitumor role.
出处
《肿瘤药学》
CAS
2017年第2期152-158,共7页
Anti-Tumor Pharmacy
基金
北京市中医管理局中西医结合项目(2014-ZYJ09)
首都医科大学附属北京世纪坛医院院国青资助课题(2015-QB02)
