姜黄素通过调控miR-100-3p/SOAT1通路对结直肠癌SW480细胞增殖和凋亡的影响  

Effect of curcumin on the proliferation and apoptosis of colorectal cancer SW480 cells by regulating MiR-100-3p/SOT1 pathway

作　　者：吴丹 张锡坚 陈伟 覃振明[3] 王函 WU Dan;ZHANG Xijian;CHEN Wei;QIN Zhenming;WANG Han(Department of Clinical Laboratory,Dongguan Traditional Chinese Medicine Hospital,Dongguan 523000,Guangdong,China;Department of Laboratory,Dongguan Nancheng Hospital,Dongguan 523000,Guangdong,China;Department of Gastroenterology,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,Guangdong,China;Department of Hematology Oncology,Dongguan Traditional Chinese Medicine Hospital,Dongguan 523000,Guangdong,China)

机构地区：[1]东莞市中医院检验科,广东东莞523000 [2]东莞市南城医院检验科,广东东莞523000 [3]广州医科大学附属第一医院消化内科,广东广州510120 [4]东莞市中医院血液肿瘤科,广东东莞523000

出　　处：《贵州医科大学学报》2025年第4期545-554,共10页Journal of Guizhou Medical University

基　　金：广东省卫生健康委员会科研项目(20221163);东莞市社会发展科技2023年重点项目(20231800940042)。

摘　　要：目的探讨姜黄素(curcumin,Cur)通过调节miR-100-3p/SOAT1通路对结直肠癌(colorectal cancer,CRC)细胞增殖和凋亡的影响。方法将结直肠癌SW480细胞分为对照组(control组)、Cur组、Cur+inhibitor NC组、Cur+miR-100-3p inhibitor组、Cur+miR-NC组及Cur+miR-100-3p mimic组;采用qRT-PCR检测miR-100-3p、SOAT1 mRNA表达水平,采用CCK-8检测24和48 h时细胞增殖能力(酶标仪测定OD 450值),Transwell实验检测细胞迁移和侵袭,流式细胞仪检测细胞凋亡,Matrigel胶三维培养观察血管拟态(vascular mimicry,VM)结构数量,Western blot检测E-钙黏蛋白(epithelial cadherin,E-cadherin)、N-钙黏蛋白(neural cadherin,N-cadherin)、波形蛋白中间丝蛋白(vimentin intermediate filament protein,vimentin)、半胱氨酸蛋白酶3(cysteine proteinase-3,cleaved caspase-3)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、固醇O-酰基转移酶1(sterol o-acyltransferase 1,SOAT1)蛋白表达水平及基质金属蛋白酶9(matrix metalloproteinase 9,MMP-9),荧光素酶活性分析miR-100-3p和SOAT1的靶向关系。结果与Control组比较,Cur组SW480细胞miR-100-3p表达水平升高,SOAT1 mRNA表达水平降低,细胞OD 450值(24、48 h)降低,迁移和侵袭个数降低,凋亡率升高,VM结构数量降低,E-cadherin及cleaved caspase-3蛋白表达水平升高,N-cadherin、vimentin、PCNA、MMP-9及SOAT1蛋白表达水平降低,差异均有统计学意义(P<0.05);下调miR-100-3p表达可减弱Cur对结直肠癌SW480细胞恶性生物学行为的抑制作用(P<0.05),上调miR-100-3p表达可进一步增强Cur对结直肠癌SW480细胞恶性生物学行为的抑制作用(P<0.05)。结论Cur可调控miR-100-3p/SOAT1通路来抑制SW480细胞恶性生物学行为。Objective To explore the effect of curcumin(Cur)on the proliferation and apoptosis of colorectal cancer(CRC)cells by regulating miR-100-3p/SOAT1 pathway.Methods CRC SW480 cells were divided into control,Cur,Cur+inhibitor NC,Cur+miR-100-3p inhibitor,Cur+miR-NC and Cur+miR-100-3p mimic groups.qRT-PCR was used to determine the expression levels of miR-100-3p and SOAT1 mRNA.CCK8 assay was applied to examine cell proliferation capacity at 24 and 48 h(using a microplate reader to read OD450 value).Cell migration and invasion were detected by Transwell assay.Cellular apoptosis was detected by flow cytometry.The number of VM structures was observed by Matrigel three-dimensional culture.Western blot was applied to detect the protein expression levels of E-cadherin,N-cadherin,vimentin,caspase-3,proliferating cell nuclear antigen(PCNA),sterol O-acyltransferase(SOAT1)and matrix metalloproteinase 9(MMP-9).Luciferase activity was used to analyze targeting relationship between miR-100-3p and SOAT1.Results When compared to control group,Cur group exhibited the increase in miR-100-3p expression level,the decreases in SOAT1 mRNA expression level,OD450 value of cells(at 24 and 48 h)and the cell numbers of migration and invasion,the increase in apoptosis rate,the decrease in the number of VM structures,the increases in the expression levels of E-cadherin and cleaved caspase-3 proteins,the decreases in the expression levels of N-cadherin,vimentin,PCNA,MMP-9 and SOAT1 proteins(P<0.05).Downregulating miR-100-3p expression weakened the inhibitory effect of Cur on the malignant biological behavior of CRC SW480 cells(P<0.05),while upregulating miR-100-3p expression enhanced the inhibitory effect of Cur on the malignant biological behavior of CRC SW480 cells(P<0.05).Conclusion Cur may regulate miR-100-3p/SOAT1 pathway to inhibit the malignant biological behavior of CRC cells.

关 键 词：姜黄素 miR-100-3p/SOAT1通路 结直肠肿瘤 细胞增殖 细胞凋亡 

分 类 号：R735.3[医药卫生—肿瘤]