基于CADD探究新疆胀果甘草查尔酮A对宫颈癌细胞增殖、凋亡作用及其分子机制  

Exploring the effect of licochalcone A from Xinjiang Glycyrrhiza inflata Bat.on proliferation and apoptosis of cervical cancer cells and its molecular mechanism based on CADD

作　　者：艾孜提艾力·艾海提 杨争[1] 木合布力·阿布力孜[1] 米热古丽·买买提明 赛力克阿拉·阿里汗 玉苏普瓦吉木·阿力木江 AIZITIAILI·Aihaiti;YANG Zheng;MOURBOUL·Ablise;MIHRIGUL·Mamtimin;SAILIKEALA·Alihan;YUSUPUWAJIMU·Alimujiang(College of Pharmacy,Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830011

出　　处：《天然产物研究与开发》2022年第3期359-367,共9页Natural Product Research and Development

基　　金：国家自然科学基金(81960625);新疆天然药物活性组分与释药技术重点实验室(XJDX1713)。

摘　　要：以新疆胀果甘草查尔酮A(licochalcone A,LicoA)为物质基础,研究其对宫颈癌细胞的增殖抑制活性、促凋亡作用及对周期的影响,并对其分子机制进行初探。从新疆胀果甘草中提取LicoA单体成分,通过1H NMR、13 C NMR及HR-EI-MS进行结构表征;通过MTT法检测在不同浓度下LicoA对人宫颈癌细胞(SiHa和HeLa)的抑制率并计算IC 50值,选取SiHa细胞为研究对象,采用流式细胞术,以AnnexinV-FITC/PI双染法检测细胞凋亡率,并测定对细胞周期的影响。通过CADD法预测LicoA作用靶点,荧光定量RT-PCR法检测宫颈癌肿瘤干细胞标记物(Bcl-2、ALDH1A1、OCT-4、UHRF1、BIRC7、BIRC5)基因及细胞周期蛋白依赖性激酶4(cyclin-dependent kinase 4,CDK4)基因的mRNA表达量。结果显示,LicoA能显著抑制2种宫颈癌细胞增殖,且呈现显著的时间和浓度依赖性;随着LicoA浓度的增加,细胞增殖速度减慢,细胞呈皱缩形态;LicoA诱导细胞凋亡作用显著,在30μg/mL时,SiHa细胞凋亡率达52.0%;LicoA可能将SiHa细胞的增殖周期阻滞在S期和G 2/M期;分子对接结果显示对CDK4蛋白有较好结合能力,预测可能具有较强的抑制作用;LicoA显著下调宫颈癌肿瘤干细胞标记物(Bcl-2、ALDH1A1、OCT-4、UHRF1、BIRC7、BIRC5)的表达量,同时抑制周期相关基因CDK4的mRNA表达。LicoA抑制宫颈癌细胞增殖的机制可能是通过将SiHa细胞的增殖周期阻滞在S期和G 2/M期,诱导细胞凋亡及抑制细胞分化。In this study,licochalcone A(LicoA)was used as the substance basis to study its proliferation inhibitory activity,apoptotic and cell cycle arresting effect on cervical cancer cells,and its molecular mechanism was initially investigated.LicoA was purified from Xinjiang Glycyrrhiza inflata Bat.,and its chemical structure was identified by 1H NMR,13 C NMR and HR-EI-MS methods.The inhibition rate of LicoA on human cervical cancer cells(SiHa and HeLa)at different concentrations was detected by MTT assay and IC 50 value was calculated.SiHa cells were selected as research object,flow cytometry was used to detect the apoptosis rate of cells by Annexinv-FITC/PI double staining,and the effect on cell cycle was determined.The CADD method was used to predict the possible target of LicoA.The mRNA expression levels of marker(Bcl-2,ALDH1A1,OCT-4,UHRF1,BIRC7,BIRC5)and cyclin-dependent kinase 4(cyclin-dependent kinase 4,CDK4)were detected by fluorescence qRT-PCR method.The results showed that LicoA could significantly inhibit the proliferation of the two cervical cancer cells in a time-dependent and concentration-dependent manner.With the increase of LicoA concentration,the cell proliferation rate slowed,and the cells showed a shrunken form.LicoA induced apoptosis significantly,and the apoptosis rate of SiHa cells reached 52.0%at 30μg/mL.LicoA may block the proliferation cycle of SiHa cells in S and G 2/M phase.Molecular docking results showed that LicoA had a good binding ability to CDK4 protein,and predicted that LicoA might have a strong inhibitory activity.LicoA significantly down-regulated the expression of stem cell markers Bcl-2,ALDH1A1,OCT-4,UHRF1,BIRC7 and BIRC5,and inhibited the mRNA expression of CDK4.The mechanism that LicoA can inhibit the proliferation of SiHa may be by arresting the proliferation cycle of SiHa in S phase and G 2/M phase,inducing cell apoptosis,and inhibiting cell differentiation.

关 键 词：甘草查尔酮A 宫颈癌细胞 增殖抑制活性 流式细胞术 CADD 肿瘤干细胞标记物 

分 类 号：R931.6[医药卫生—生药学]