基于PTEN/PI3K/AKT信号通路探讨隐丹参酮抑制结直肠癌的作用机制  

Discussion on the mechanism of cryptotanshinone inhibiting colorectal cancer based on PTEN/PI3K/AKT signaling pathway

作　　者：冯叶雯 安庆文 杨楚琪 吴梦婷 姜涛 张光霁[1,2,3] FENG Yewen;AN Qingwen;YANG Chuqi;WU Mengting;JIANG Tao;ZHANG Guangji(School of Basic Medical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;Key Laboratory of‘Bloodstasis-toxin’Syndrome of Zhejiang Province,Hangzhou 310053,China;Traditional Chinese Medicine‘Preventing Disease’Wisdom Health Project Research Center of Zhejiang Province,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学基础医学院,杭州310053 [2]浙江省中医“瘀毒”证重点实验室,杭州310053 [3]中医“治未病”智慧健康浙江省工程研究中心,杭州310053

出　　处：《中华中医药杂志》2024年第12期6666-6670,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：浙江省中医药现代化专项(No.2020ZX005);国家自然科学基金青年科学基金项目(No.82204950)。

摘　　要：目的:研究传统中药丹参提取物隐丹参酮抑制结直肠癌Caco-2细胞的作用机制。方法:通过CCK8法检测隐丹参酮对Caco-2细胞活性的影响;划痕实验检测隐丹参酮对Caco-2细胞迁移能力的影响;平板集落实验、EdU染色和免疫荧光标记Ki67检测隐丹参酮对Caco-2细胞增殖能力的影响;Western Blot检测隐丹参酮对Caco-2细胞中增殖相关蛋白增殖细胞核抗原(PCNA),细胞周期相关蛋白细胞周期蛋白D1(cyclin D1)、周期蛋白依赖性激酶4(CDK4)、周期蛋白依赖性激酶6(CDK6)以及磷酸酶与张力蛋白同源物(PTEN)/磷脂酰肌醇-3激酶(PI3K)/蛋白激酶(AKT)信号通路相关蛋白表达的影响。结果:隐丹参酮抑制Caco-2细胞活力,并呈一定的剂量和时间依赖性。与0μg/mL隐丹参酮组比较,10、15、20μg/mL隐丹酮组显著抑制Caco-2细胞迁移能力和增殖能力(P<0.05,P<0.01);20μg/mL隐丹参酮组显著降低Caco-2细胞中增殖相关蛋白PCNA表达(P<0.05),15、20μg/mL隐丹参酮组细胞周期相关蛋白cyclin D1、CDK4、CDK6表达显著降低(P<0.01,P<0.05);20μg/mL隐丹参酮组调控Caco-2细胞中PTEN/PI3K/AKT信号通路相关蛋白表达(P<0.01)。结论:隐丹参酮能够显著抑制结直肠癌细胞增殖,其抗增殖能力可能与调控PTEN/PI3K/AKT信号通路,调节细胞周期蛋白表达有关。Objective:To investigate the mechanism of cryptotanshinone,a traditional Chinese medicine extract of Salviae Miltiorrhizae Radix Et Rhizoma,in inhibiting colorectal cancer Caco-2 cells.Methods:The effect of cryptotanshinone on the activity of Caco-2 cells was detected by CCK8 method.The effect of cryptotanshinone on migration ability of Caco-2 cells was detected by scratch test.The effects of cryptotanshinone on the proliferation of Caco-2 cells were detected by plate colony assay,EdU staining and immunofluorescence labeling Ki67.Western Blot was used to detect the effect of cryptotanshinone on the expression of proliferation-related protein proliferating cell nuclear antigen(PCNA),cell cycle-related protein cyclin D1,cyclindependent kinase 4(CDK4),cyclin-dependent kinase 6(CDK6)and PTEN/PI3K/AKT signaling pathway related proteins in Caco-2 cells.Results:Cryptotanshinone inhibited Caco-2 cell viability in a dose-dependent and time-dependent manner.Compared with 0μg/mL cryptotanshinone,10,15,20μg/mL cryptotanshinone significantly inhibited the migration and the proliferation of Caco-2 cells(P<0.05,P<0.01),20μg/mL cryptotanshinone could significantly reduce the expressions of proliferation-related proteins PCNA(P<0.05),15,20μg/mL cryptotanshinone could significantly reduce the protein expression of cyclin D1,CDK4 and CDK6 in Caco-2 cells(P<0.01,P<0.05);20μg/mL cryptotanshinone could regulate the expression of PTEN/PI3K/AKT signaling pathway in Caco-2 cells(P<0.01).Conclusion:Cryptotanshinone can significantly inhibit the proliferation of colorectal cancer cells,and its anti-proliferation ability may be related to the regulation of PTEN/PI3K/AKT signaling pathway and the regulation of cell cycle-related protein.

关 键 词：隐丹参酮 结直肠癌 增殖 磷酸酶与张力蛋白同源物 磷脂酰肌醇-3激酶 蛋白激酶 机制 细胞周期蛋白 

分 类 号：R73[医药卫生—肿瘤]