白藜芦醇调控磷脂酰肌醇-3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白通路抑制人结肠癌细胞SW480增殖和迁移促进凋亡的研究  

Resveratrol inhibits proliferation,migration and promotes apoptosis of human colon cancer SW480 cells via PI3K/Akt/mTOR pathway

作　　者：杨文聪 王雪婷 Yang Wencong;Wang Xueting(Internal Medicine,Jinhua Maternal and Child Health Hospital,Jinhua,Zhejiang 321000,China)

机构地区：[1]金华市妇幼保健院内科,浙江金华321000

出　　处：《中国药物与临床》2024年第16期1083-1088,共6页Chinese Remedies & Clinics

摘　　要：目的探讨白藜芦醇对人结肠癌细胞SW480增殖、迁移的抑制作用和对其凋亡的促进作用,并分析对磷脂酰肌醇-3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/Akt/mTOR)通路的调控作用。方法2023年1月取人结肠癌细胞株SW480,培养、传代,生长至对数期时将其分组,共设置空白对照组(BC组)、阳性对照组(PC组)、白藜芦醇低、中、高剂量组(LD组、MD组、HD组),每组设有3个复孔。白藜芦醇LD、MD、HD组分别给予50、100、200μmol/L白藜芦醇干预,PC组给予12.5 mg/L奥沙利铂干预,BC组给予等量无菌0.9%氯化钠注射液干预,继续培养48 h。以光学显微镜观察细胞形态学变化;以细胞钙黄绿素法(MMT)检测细胞增殖,以细胞划痕实验检测细胞迁移,以流式细胞仪检测细胞凋亡,以实时荧光定量聚合酶链反应(PCR)(RT-qPCR)法检测磷脂酰肌醇-3-激酶(PI3K)、蛋白激酶B(Akt)、哺乳动物雷帕霉素靶蛋白(mTOR)mRNA相对表达量,以蛋白免疫印迹法检测PI3K、Akt、mTOR、P21蛋白、细胞周期蛋白D1(CyclinD1)蛋白相对表达量及Akt、mTOR磷酸化水平。结果BC组细胞形态学无明显变化,其余4组均出现不同程度细胞皱缩、变圆、体积缩小、折光性和贴壁能力减弱、细胞脱落等情况,且PC组可见少量裂解碎片,MD组可见明显裂解碎片,HD组可见较多裂解碎片;增殖抑制率对比,HD组最高,MD组次之,PC组稍低,LD组更低;划痕区域相对宽度对比,BC组最宽,LD组次之,PC组稍窄,MD组更窄,HD组最窄;细胞凋亡率对比,HD组最高,MD组次之,PC组稍低,LD组更低,BC组最低;PI3K mRNA及蛋白、P21、CyclinD1蛋白相对表达量,p-Akt、p-mTOR对比,BC组均最高,LD组均次之,PC组均稍低,MD组均更低,HD组均最低;P21蛋白相对表达量对比,HD组最高,MD组次之,PC组稍低,LD组更低,BC组最低。上述指标每2组间对比差异均有统计学意义(P<0.05)。结论白藜芦醇可促使人结肠癌细胞SW480形态学变化、裂解,抑�Objective To investigate the inhibitory effect of resveratrol on proliferation,migration and apoptosis of human colon cancer cell line SW480,and to analyze the regulatory effect of resveratrol on the PI3K/Akt/mTOR pathway.Methods Human colon cancer cell line SW480 was cultured,passaged,and grouped into three groups at the logarithmic phase.The blank control group(BC group),positive control group(PC group),and low/medium/high dose resveratrol group(LD/MD/HD group)were set up with three compound holes.Resveratrol LD,MD and HD groups were given 50,100 and 200μmol/L of resveratrol intervention.The PC group was given 12.5 mg/L of oxaliplatin intervention,and the BC group was given the same amount of sterile saline intervention.All of these groups were then cultured for 48 hours.The morphological changes of the cells were observed by light microscopy.Cell proliferation was detected by MMT assay.Cell migration was detected by Transwell chamber mi-gration test.Cell apoptosis was detected by flow cytometry.The relative expression of PI3K,Akt,and mTOR was detected by RT-qPCR.The relative expression of PI3K,Akt,mTOR,P21,CyclinD1,and the phosphorylation of Akt and mTOR were detected by Western blot.Results There were no obvious changes in cell morphology in the BC group.Cell shrinkage,roundness,volume reduction,refraction,and adherence weakening,cell exfoliation were observed in the other four groups.A few fragments were observed in the PC group,obvious fragments in the MD group,and more fragments in the HD group.The inhibition rate of proliferation in the HD group was the highest,followed by the MD group,the PC group was slightly lower and the LD group was lower.The relative width of scratch area was the widest in the BC group,followed by the LD group,slightly narrower in the PC group,narrow-er in the MD group and narrowest in the HD group.The apoptotic rate of the HD group was the highest,followed by the MD group;the PC group was slightly lower,the LD group was lower,and the BC group was the lowest.The relative expression

关 键 词：结肠肿瘤 细胞凋亡 白藜芦醇 

分 类 号：R285[医药卫生—中药学]