异鼠李素通过nSMase2依赖的外泌体生成途径抑制结肠癌肝转移的作用机制初探  

A Preliminary Investigation on Mechanism of Isorhamnetin Inhibiting Colon Cancer Liver Metastasis through nSMase2-dependent Exosome Production Pathway

作　　者：李昊泽 周晶 季青[1] LI Haoze;ZHOU Jing;JI Qing(Department of Medical Oncology and Cancer Institute,Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院肿瘤科/肿瘤研究室,上海201203

出　　处：《辽宁中医杂志》2024年第9期9-12,I0001,共5页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82074225,82274297)。

摘　　要：目的探讨中药活性成分异鼠李素(isorhamnetin,ISO)对结肠癌肝转移和肿瘤外泌体分泌的干预作用,并进一步揭示其潜在作用机制。方法使用CCK-8法检测ISO对小鼠结肠癌细胞MC38细胞增殖的影响。使用NanoSightNS300系统检测ISO对MC38细胞外泌体释放的影响。通过实时荧光定量检测系统(quantitative PCR detecting system,qPCR)检测ISO对MC38细胞对中性鞘磷脂酶-2(neutral sphingomyelinase 2,nSMase2)、Rab27a等外泌体生成、分泌相关基因表达的影响。将12只C57BL/6小鼠随机分为对照组、MC38-外泌体组、ISO组、阳性对照组(外泌体抑制剂GW4869组),造模4周后检测小鼠肝脏转移灶数量及肝脏重量。通过qPCR检测肝转移灶中nSMase2、Rab27a等外泌体生成、分泌相关基因的表达情况。结果ISO能够抑制MC38细胞增殖,且在20μmol/L浓度以下时,抑制率低于10%,为无毒剂量;ISO能够减少MC38细胞外泌体的分泌,且具有时间依赖性(P<0.05);ISO能够抑制MC38细胞中外泌体生成相关基因nSMase2 mRNA的表达(P<0.001),而对外泌体分泌相关基因表达无明显抑制作用。与对照组比较,ISO组小鼠的肝脏重量明显降低(P<0.05),转移瘤数目明显减少;ISO能明显抑制小鼠结肠癌肝转移灶中外泌体生成相关基因nSMase2 mRNA的表达(P<0.001)。结论ISO可能通过nSMase2依赖途径抑制小鼠结肠癌细胞外泌体生成,进而抑制结肠癌肝转移。Objective To investigate the interventional effects of isorhamnetin(ISO),the active ingredient of traditional Chinese medicine,on hepatic metastasis of colon cancer and tumor-associated exosome secretion and to further identify its potential mechanism.Methods The effect of ISO on the proliferation of colon cancer MC38 cells in mice was detected using Cell Counting Kit 8(CCK8).The effect of ISO on exosome release from MC38 cells was detected by using NanoSight NS300 instrument.And Quantitative PCR Detecting System(qPCR)was performed to detect the effect of ISO on the expressions of genes related to the production and secretion of exosomes such as(neutral sphingomyelinase 2,nSMase2)and Rab27a in MC38 cells.Twelve C57BL/6 mice were randomly divided into the control group,MC38-exosomes group,ISO group and positive control group(exosome inhibitor GW4869 group).And the number of liver metastases and liver weight were measured four weeks after modeling.And qPCR was performed to detect the expressions of genes related to the production and secretion of exosomes such as neutral sphingomyelinase 2(nSMase2)and Rab27a in liver metastases.Results ISO can inhibit the proliferation of MC38 cells,with inhibition rate of less than 10%at concentrations below a non-toxic dose of 20μmol/L.Besides,ISO can reduce the secretion of exosomes in MC38 cells in a time-dependent manner(P<0.05),and it also can inhibit the expression of nSMase2 mRNA,a gene related to exosome production,in MC38 cells(P<0.001).But it did not have the same inhibitory effect on the expressions of genes related to exosome secretion.Compared with that of the control group,the liver weight of mice in the ISO group was significantly reduced(P<0.05).The number of metastases was also significantly reduced.And ISO significantly inhibited the expression of nSMase2 mRNA,a gene related to exosome production,in liver metastases of mice with colon cancer(P<0.001).Conclusion ISO may inhibit exosome production in colon cancer cells of mice through the nSMase2-dependent pathway

关 键 词：异鼠李素 结肠癌 肝转移 外泌体 nSMase2 

分 类 号：R285[医药卫生—中药学]