二十二碳六烯酸抑制人结肠癌细胞系HT-29增殖  

Docosahexaenoic acid inhibits proliferation of human colon cancer cell line HT-29

作　　者：姚安军 陈凌子 金惠仙 YAO Anjun;CHEN Lingzi;JIN Huixian(Department of Clinical Nutrition,Affiliated Jinhua Hospital,Zhejiang University School of Medicine,Jinhua 321000;Department of Laboratory Medicine,Wucheng District People′s Hospital,Jinhua 321000,China)

机构地区：[1]浙江大学医学院附属金华医院临床营养科,浙江金华321000 [2]金华市婺城区人民医院检验科,浙江金华321000

出　　处：《基础医学与临床》2024年第8期1107-1112,共6页Basic and Clinical Medicine

基　　金：浙江省医药卫生科技计划项目(2022KY1329);金华市科技计划项目社会发展类重点项目(2020-3-034);金华市科技计划项目公益类项目(2021-4-120)。

摘　　要：目的探讨二十二碳六烯酸(DHA)对人结肠癌细胞系HT-29的影响及其作用机制。方法将人结肠癌细胞系HT-29分为对照组,25、50和100μmol/L DHA处理组,以及100μmol/L DHA+30μmol/L 740Y-P处理组。MTT检测HT-29细胞增殖,annexin V-FITC/PI检测细胞凋亡,Western blot检测Bcl-2、Bax蛋白以及p-PI3K、p-Akt和p-mTOR蛋白表达,RT-qPCR检测NLRP3、Caspase-1和IL-1βmRNA相对表达量。结果与对照组相比,DHA 25、50和100μmol/L处理HT-29细胞后细胞存活率下降(P<0.05或P<0.01),细胞凋亡率增加(P<0.05),Bcl-2/Bax比值降低(P<0.05),PI3K、Akt和mTOR的磷酸化水平均下降(P<0.05或P<0.01),NLRP3、Caspase-1和IL-1βmRNA相对表达量均下降(P<0.05或P<0.01)。此外,DHA 100μmol/L和740Y-P 30μmol/L共同处理HT-29细胞后,细胞生存率、蛋白磷酸化水平(p-PI3K、p-Akt和p-mTOR)、mRNA相对表达量(NLRP3、Caspase 1和IL-1β)均低于对照组(P<0.05)和740Y-P 30μmol/L组(P<0.05),而高于DHA 100μmol/L组(P<0.05或P<0.01)。结论DHA抑制人结肠癌细胞系HT-29增殖,作用机制可能与抑制PI3K/Akt/mTOR和NLRP3/Caspase-1/IL-1β信号分子通路有关。Objective To investigate the effect of docosahexaenoic acid(DHA)on human colon cancer cell line HT-29 and underlying mechanism.Methods Human colon cancer cell line HT-29 was incubated with DMSO(control),DHA(25,50,100μmol/L)and 100μmol/L DHA and/or 30μmol/L 740Y-P.Proliferation was examined by MTT;apoptosis was detected by annexin V-FITC/PI.Western blot was used for detection of protein expression of Bcl-2,Bax apoptosis-related protein and PI3K/Akt/mTOR pathway,and RT-qPCR was used for checking mRNA expression of NLRP3/Caspase-1/IL-1βpathway.Results Compared with the control group,DHA 25,50,and 100μmol/L treatment of HT-29 cells resulted in decreased cell survival(P<0.05),increased apoptosis(P<0.05),decreased Bcl-2/Bax ratio(P<0.05)and decreased phosphorylation of PI3K,Akt and mTOR in HT-29 cells(P<0.05 or P<0.01).Expressions of NLRP3,Caspase-1 and IL-1βmRNA were decreased(P<0.05).In addition,cell viability,protein phosphorylation(p-PI3K,p-Akt,p-mTOR)and relative mRNA expression of NLRP3,Caspase 1,and IL-1βwere lower in HT-29 cells which were co-incubated with DHA 100μmol/L and 740Y-P 30μmol/L than those in the control group(P<0.05 or P<0.01)and 740Y-P 30μmol/L group(P<0.05),while higher than that of DHA 100μmol/L group(P<0.05 or P<0.01).Conclusions DHA inhibits the proliferation of human colon cancer cell line HT-29,its mechanism is potentially related to the inhibition of PI3K/Akt/mTOR and NLRP3/Caspase-1/IL-1βsignaling pathways.

关 键 词：二十二碳六烯酸 PI3K/AKT/MTOR NOD样受体pyrin结构域相关蛋白3(NLRP3) 结肠癌 细胞增殖 

分 类 号：R735[医药卫生—肿瘤]