柚皮素通过PI3K/Akt信号通路促进人胃癌SGC-7901细胞凋亡的实验研究  

EXPERIMENTAL STUDY OF NARINGENIN IN PROMOTING APOPTOSIS OF HUMAN GASTRIC CANCER SGC-7901 CELLS THROUGH PI3K/AKT SIGNALING PATHWAY

作　　者：陈瑶莉 吴彩霞 Chen Yaoli;Wu Caixia(Department of Gastroenterology,People′s Hospital of Tongchuan,Tongchuan 727000,China;不详)

机构地区：[1]陕西省铜川市人民医院消化内科,727000 [2]咸阳市中心医院检验科

出　　处：《中国煤炭工业医学杂志》2020年第5期449-456,共8页Chinese Journal of Coal Industry Medicine

基　　金：陕西省教育厅自然科学研究项目(编号:2016JK2031)。

摘　　要：目的研究柚皮素对人胃癌SGC-7901细胞凋亡和PI3K/Akt信号通路的影响,并探讨其可能作用机制。方法体外培养人胃癌SGC-7901细胞,采用不同浓度的柚皮素0、20、40、80μmol/L作用24、48、72h,各浓度梯度设置6个复孔,应用MTT法检测柚皮素对人胃癌SGC-7901细胞增殖的影响,应用倒置显微镜观察细胞形态学变化,应用流式细胞仪检测48h人胃癌SGC-7901细胞凋亡情况,并应用RT-PCR技术和Western blot技术检测人胃癌SGC-7901细胞凋亡相关因子Bax、Bcl-2、Caspase-3、Caspase-9的表达水平以及PI3K/Akt信号通路情况。结果MTT检测结果显示,20、40、80μmol/L柚皮素处理人胃癌SGC-7901细胞24、48、72h后,与阴性对照组(0μmol/L柚皮素)比较,不同浓度柚皮素均可明显抑制人胃癌SGC-7901细胞增殖,且表现为时间和浓度依赖性的抑制作用(P<0.05);倒置显微镜检测结果显示,阴性对照组人胃癌SGC-7901细胞生长状态良好,细胞呈多边形或梭形贴壁生长,而不同浓度柚皮素处理SGC-7901细胞48h后,细胞松散,形态不规则,大部分细胞皱缩、破裂,部分细胞呈现圆形固缩,生长变慢,细胞数量也明显减少,并表现为浓度依赖性;流式细胞技术检测结果显示,随着柚皮素浓度的增加,人胃癌SGC-7901细胞凋亡率随之升高,不同浓度柚皮素处理的SGC-7901细胞48h检测凋亡率显著高于阴性对照组(P<0.05);RTPCR和Western blot检测结果显示,各浓度组SGC-7901细胞凋亡相关因子Bax、Caspase-3以及Caspase-9mRNA和蛋白表达水平显著高于阴性对照组(P<0.05),Bcl-2mRNA和蛋白表达水平显著低于阴性对照组(P<0.05),且各浓度组呈递增或递减趋势(P<0.05);RT-PCR检测结果显示,各浓度组SGC-7901细胞PI3K、Akt mRNA表达水平显著低于阴性对照组(P<0.05),Western blot检测结果显示,各浓度组SGC-7901细胞p-PI3K、p-Akt蛋白表达水平显著低于阴性对照组(P<0.05),表现为20μmol/L柚皮素组>40μmol/L柚皮素组>80μmol/L柚皮�Objective To study the effects of Naringenin on apoptosis of human gastric cancer SGC-7901 cells and PI3 K/Akt signaling pathway,and to explore its possible role mechanism.Methods Human gastric cancer SGC-7901 cells were cultured in vitro and treated with different concentrations of Naringenin at 0,20,40,80μmol/L for 24,48 and 72 h.Each concentration gradient was set up with 6 multiple wells.Effects of Naringenin on the proliferation of human gastric cancer SGC-7901 cells were detected by MTT.Inverted microscope was used to observe cell morphology changes,and flow cytometry was used to detect the apoptosis of 48 h human gastric cancer SGC-7901 cells,and the RT-PCR and Western blot were used to detect the expression levels of apoptosis-related molecules(Bax,Bcl-2,Caspase-3,Caspase-9)and PI3 K/Akt signaling pathway in human gastric cancer SGC-7901 cells.Results The results of MTT detection showed that after 20,40 and 80μmol/L Naringenin treated human gastric cancer SGC-7901 cells for 24,48 and 72 h,compared with negative control group(0μmol/L Naringenin),different concentrations of Naringenin could significantly inhibit the proliferation of human gastric cancer SGC-7901 cells,and showed a time-and concentration-dependent inhibitory effect(P<0.05).The inverted microscope test results showed that thehuman gastric cancer SGC-7901 cells in negative control group grew in good condition,and the cells showed polygonal or spindle-shaped adherent growth.After 48 h of treatment of SGC-7901 cells with different concentrations of Naringenin,the cells were loose and irregular in shape,most of the cells shrinked and ruptured and some of the cells appeared round pyknosis and slow growth,and the number of cells was also significantly reduced,showing concentration-dependence.The results of flow cytometry showed that with the increase of Naringenin concentration,the apoptosis rate of human gastric cancer SGC-7901 cells was increased,and the apoptosis rate of SGC-7901 cells with different concentrations of Naringenin at 48 h was s

关 键 词：柚皮素 胃癌 SGC-7901 细胞凋亡 PI3K/AKT信号通路 

分 类 号：R735.2[医药卫生—肿瘤]