苦豆碱抑制膀胱癌253J细胞增殖的作用及机制研究  

Inhibitory effect and mechanism of aloperin on proliferation of bladder carcinoma253J cells

作　　者：高义胜 李敬东[2] 何相飞[1] 高磊[1] 徐纪元 左世帅 GAO Yi-sheng;LI Jing-dong;HE Xiang-fei;GAO Lei;XU Ji-yuan;ZUO Shi-shua(Department of Urology,Linyi People's Hospital Linyi 276000,Shandong Province,China;Department of Oncology,Linyi People's Hospital Linyi 276000,Shandong Province,China;Graduate Training Base,LinyiPeoples's Hospitalof Guangzhou University of Traditional Chinese Medicine,Linyi 276000,Shandong Province,China)

机构地区：[1]临沂市人民医院泌尿外科,山东临沂276000 [2]临沂市人民医院肿瘤内科,山东临沂276000 [3]广州中医药大学临沂市人民医院研究生培养基地,山东临沂276000

出　　处：《中国临床药理学杂志》2025年第5期676-680,共5页The Chinese Journal of Clinical Pharmacology

基　　金：山东省中医药科技发展计划基金资助项目(M-2022106)。

摘　　要：目的探究苦豆碱对膀胱癌细胞生长及内质网应激依赖线粒体途径的调控作用。方法将253J细胞随机分为对照组、低剂量实验组(25.0μmoL·L^(-1)苦豆碱)、中剂量实验组(50.0μmoL·L^(-1)苦豆碱)、高剂量实验组(100.0μmoL·L^(-1)苦豆碱)、4-PBA组[100.0μmoL·L^(-1)苦豆碱+20μmoL·L^(-1)4-苯基丁酸(4-PBA)]、si-NC组(转染si-NC+100.0μmoL·L^(-1)苦豆碱)、si-CHOP组[转染si-C/EBP同源蛋白(CHOP)+100.0μmoL·L^(-1)苦豆碱]。用5-乙炔基-2’-脱氧尿苷(EdU)实验检测细胞增殖,用蛋白质印迹法检测相关蛋白表达,用流式细胞术检测细胞凋亡,用JC-1法检测线粒体膜电位(MMP)水平。结果对照组、低剂量实验组、中剂量实验组、高剂量实验组EdU阳性细胞率分别为(37.51±2.98)%、(29.52±2.19)%、(23.10±2.58)%和(12.94±2.52)%,CHOP蛋白相对表达水平分别为0.29±0.04、0.53±0.05、0.71±0.08和0.90±0.12;对照组、高剂量实验组、4-PBA组细胞凋亡率分别为(4.79±0.62)%、(23.01±2.03)%和(13.75±1.14)%;对照组、高剂量实验组、si-NC、si-CHOP组细胞CHOP蛋白表达分别为0.30±0.03、0.92±0.08、0.91±0.11和0.20±0.02,细胞凋亡率分别为(4.39±0.30)%、(22.11±1.82)%、(22.65±2.40)%和(15.32±1.19)%,线粒体细胞色素C(Cyto-C)蛋白相对表达水平为0.78±0.07、0.43±0.05、0.42±0.05和0.65±0.06,细胞质Cyto-C蛋白相对表达水平为0.51±0.06、0.91±0.10、0.88±0.05和0.61±0.04,MMP水平分别为(100.00±2.79)%、(57.49±3.34)%、(54.09±4.32)%和(69.91±6.81)%;低、中、高剂量组的上述指标分别与对照组比较,4-PBA组的上述指标与高剂量实验组比较,si-CHOP组的上述指标与si-NC组比较,在统计学上差异均有统计学意义(均P<0.01)。结论苦豆碱可通过上调CHOP介导内质网应激依赖的线粒体凋亡途径抑制253J细胞增殖。Objective To investigate the regulatory effects of aloperin on bladder cancer cell growth and endoplasmic reticulum stressdependent mitochondrial pathway.Methods 253J cells were randomly divided into control group,experimental-L group(25.0μmoL·L^(-1)aloperin),experimental-M group(50.0μmoL·L^(-1)aloperin),experimental-H group(100.0μmoL·L^(-1)aloperin),4-PBA group[100.0μmoL·L^(-1)aloperin+20μmoL·L^(-1)4-phenylbutyric acid(4-PBA)],si-NC(transfected with si-NC+100.0μmoL·L^(-1)aloperin),si-CHOP group[transfected with si-CCAAT-enhancer-binding protein homologous protein(CHOP)+100.0μmoL·L^(-1)aloperin].Cell proliferation was detected by 5-ethynyl-2′-deoxyuridine(EdU)assay;protein expression was detected by Western blot assay;apoptosis was detected by flow cytometry;mitochondrial membrane potential(MMP)was detected by JC-1 assay.Results The EdU positive cell rates in control group,experimental-L group,experimental-M group and experimental-H group were(37.51±2.98)%,(29.52±2.19)%,(23.10±2.58)%and(12.94±2.52)%,respectively;the expression levels of C/EBP homologous protein(CHOP)were 0.29±0.04,0.53±0.05,0.71±0.08,0.90±0.12,respectively.The apoptosis rates in control group,experimental-H group and 4-PBA group were(4.79±0.62)%,(23.01±2.03)%and(13.75±1.14)%,respectively.The expressions of CHOP protein in control group,experimental-H,si-NC and si-CHOP groups were0.30±0.03,0.92±0.08,0.91±0.11 and 0.20±0.02,respectively;the apoptosis rates were(4.39±0.30)%,(22.11±1.82)%,(22.65±2.40)%and(15.32±1.19)%,respectively;mitochondrial cytochrome C(Cyto-C)protein levels were 0.78±0.07,0.43±0.05,0.42±0.05 and 0.65±0.06;cytoplasmic Cyto-C protein levels were0.51±0.06,0.91±0.10,0.88±0.05 and 0.61±0.04;MMP levels were(100.00±2.79)%,(57.49±3.34)%,(54.09±4.32)%and(69.91±6.81)%,respectively.The above indexes in the experimental-L,-M,-H groups were compared with the control group,the above indexes in the 4-PBA group were compared with the experimental-H group,and the above indexes in the si-CHOP group w

关 键 词：苦豆碱 膀胱癌 内质网应激 线粒体凋亡 增殖 

分 类 号：R28[医药卫生—中药学]