贝母素乙通过诱导G0/G1期阻滞抑制结肠癌细胞的增殖  

Peiminine inhibits the proliferation of colon cancer cells by inducing G0/G1 phase arrest

作　　者：孙丽丽 白冰 杨霞 李玥 李一权 韩继成[2] 房金波 李霄 尚超 朱羿龙[2] 金宁一 SUN Lili;BAI Bing;YANG Xia;LI Yue;LI Yiquan;HAN Jicheng;FANG Jinbo;LI Xiao;SHANG Chao;ZHU Yilong;JIN Ningyi(Department of Pathology and Pathophysiology,College of Medicine,Yanbian University,Yanji 136200,Jilin,China;Academician Workstation of Changchun University of Chinese Medicine,Changchun 130117,Jilin,China;Changchun Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Changchun 130122,Jilin,China)

机构地区：[1]延边大学医学院病理学与病理生理学教研室,吉林延吉136200 [2]长春中医药大学院士工作室,吉林长春130117 [3]中国农业科学研究院长春兽医研究所,吉林长春130122

出　　处：《中国肿瘤生物治疗杂志》2024年第4期326-332,共7页Chinese Journal of Cancer Biotherapy

基　　金：吉林省科技发展计划(No.20220508075RC);长春市科技发展计划项目创新平台与人才计划青年科技人才创新创业专项(No.21QC01);吉林省中医药科技项目(No.2024255)。

摘　　要：目的:探讨贝母素乙对结肠癌HCT116细胞增殖的抑制作用及其分子机制。方法:利用不同浓度的贝母素乙处理人结肠癌细胞HCT116和正常结肠上皮细胞CCD841 CON,通过CCK-8法和结晶紫染色法检测贝母素乙对HCT116和CCD841 CON细胞增殖活力的影响,流式细胞术和WB法检测贝母素乙对HCT116细胞周期及其细胞周期相关蛋白表达的影响。构建HCT116移植瘤裸鼠模型和AOM/DSS结肠癌小鼠模型,观察贝母素乙对小鼠模型肿瘤生长和OS的影响,免疫组化法和WB法检测对移植瘤或肿瘤组织中细胞周期相关蛋白CDK4、CDK6和cyclin D1表达的影响。结果:贝母素乙可显著抑制结肠癌HCT116细胞的增殖能力(P<0.01),诱导HCT116细胞周期G0/G1期阻滞(P<0.01),降低CDK4、CDK6和cyclin D1的蛋白表达水平(均P<0.01)。荷瘤小鼠实验结果显示,贝母素乙(0.75 mg/kg)显著抑制HCT116细胞移植瘤的生长并延长荷瘤裸鼠的OS(P<0.05或P<0.01),降低AOM/DSS模型小鼠的体质量、延长OS、减少癌变肠组织的肿瘤个数和肿瘤体积,下调肿瘤组织中CDK4、CDK6和cyclin D1的蛋白表达(P<0.01或P<0.05)。结论:贝母素乙通过下调CDK4、CDK6和cyclin D1的表达水平,引起细胞周期G0/G1期阻滞,从而抑制结肠癌HCT116细胞的增殖。Objective:To explore the inhibitory effects and molecular mechanism of peiminine on colon cancer HCT116 cells.Methods:Human colon cancer HCT116 cells and normal colon epithelial CCD841 CON cells were treated with various concentrations of peiminine.The effects of peiminine on the proliferative vitality of HCT116 and CCD841 CON cells were detected by CCK-8 method and crystal violet staining.The effects of peiminine on the cell cycle and the expression of cycle related proteins in HCT116 cells were detected by flow cytometry and Western blot.HCT116 nude mouse transplantation tumor model and the AOM/DSS-induced colon cancer mouse model were constructed to observe the effects of peiminine on tumor growth and overall survival in the mouse model.The effects of peiminine on the expressions of cell cycle related proteins CDK4,CDK6 and cyclin D1 in transplanted tumors or tumor tissues were detected by immunohistochemistry and Western blot.Results:Peiminine significantly inhibited the proliferation ability(P<0.01),induced the G0/G1 phase arrest(P<0.01),and reduced the expression levels of CDK4,CDK6,and cyclin D1 proteins in HCT116 cells(all P<0.01).The results of tumor bearing nude mice showed that peiminine(0.75 mg/kg)significantly inhibited the growth of HCT116 cell transplanted tumors,increased the overall survival rate(P<0.05 or P<0.01).Peiminine also reduced the weight and prolonged the overall survival of AOM/DSS model mice,reduced the number and volume of tumors in cancerous colon tissues,and down-regulated the expressions of CDK4,CDK6 and cyclin D1 proteins in tumor tissues(P<0.05 or P<0.01).Conclusion:Peiminine can cause cell cycle G0/G1 phase arrest,and thus suppress the proliferation of colon cancer HCT116 cells through downregulating the expression levels of CDK4,CDK6,and cyclin D1.

关 键 词：贝母素乙 结肠癌 HCT116细胞 G0/G1期阻滞 增殖 

分 类 号：R735.35[医药卫生—肿瘤] R730.2[医药卫生—临床医学] R730.52