南蛇藤提取物通过调控微小RNA-378对结肠癌细胞生物学行为的影响  

Effect of celastrus orbiculatus extract on cell biological behaviors in colon cancer by regulating microRNA-378

作　　者：王威 邓瑞 周培华[1] WANG Wei;DENG Rui;ZHOU Peihua(Department of Gastrointestinal Surgery,Sinopharm Dongfeng General Hospital Affiliated to Hubei University of Medicine,Shiyan,Hubei 442000;Electrocardiogram Room,Sinopharm Dongfeng General Hospital Affiliated to Hubei University of Medicine,Shiyan,Hubei 442000)

机构地区：[1]湖北医药学院附属国药东风总医院胃肠外科,湖北十堰442000 [2]湖北医药学院附属国药东风总医院心电图室,湖北十堰442000

出　　处：《河北中医》2024年第12期2017-2023,共7页Hebei Journal of Traditional Chinese Medicine

摘　　要：目的观察探讨南蛇藤提取物对结肠癌Caco-2细胞生物学行为的影响及其可能作用机制。方法将结肠癌Caco-2细胞分为低、中、高剂量组及阳性对照组、对照组、miR-NC组、miR-378组、anti-miR-NC+高剂量组、anti-miR-378+高剂量组,其中低、中、高剂量组分别用20、60、180μg/mL南蛇藤提取物干预,阳性对照组用环磷酰胺30μmol/L干预,对照组正常培养。miR-NC组、miR-378组分别用脂质体将miR-NC、miR-378 mimcs分别转染至Caco-2细胞,anti-miR-NC+高剂量组、anti-miR-378+高剂量组分别用脂质体将anti-miR-NC、anti-miR-378转染至Caco-2细胞中,并加入含有180μg/mL南蛇藤提取物的培养基干预。每组细胞设置9个复孔。MTT法、流式细胞术、Transwell小室实验分别检测细胞增殖、凋亡、迁移及侵袭,Western blot法检测Ki67、基质金属蛋白酶2(MMP-2)、MMP-9、Cleaved-caspase-3蛋白表达量,实时荧光定量聚合酶链式反应(qRT-PCR)法检测miR-378 mRNA表达量。结果与对照组比较,南蛇藤提取物低、中、高剂量组细胞增殖率和Ki67、MMP-2、MMP-9蛋白表达水平均降低(P<0.05),迁移及侵袭细胞数减少(P<0.05),细胞凋亡率、Cleaved-caspase-3蛋白表达水平和miR-378 mRNA表达量均升高(P<0.05),且呈剂量依赖性(P<0.05)。与miR-NC组比较,miR-378组细胞增殖率和Ki67、MMP-2、MMP-9蛋白表达水平均降低(P<0.05),迁移及侵袭细胞数减少(P<0.05),细胞凋亡率、Cleaved-caspase-3蛋白表达水平和miR-378 mRNA表达量均升高(P<0.05)。与anti-miR-NC+高剂量组比较,anti-miR-378+高剂量组细胞增殖率和Ki67、MMP-2、MMP-9蛋白表达水平均升高(P<0.05),迁移及侵袭细胞数增多(P<0.05),凋亡率和Cleaved-caspase-3蛋白表达水平、miR-378 mRNA表达均降低(P<0.05)。结论南蛇藤提取物可通过促进miR-378表达来抑制结肠癌细胞恶性生物学行为。Objective To explore the effect of celastrus orbiculatus extract on the biological behaviors of colon cancer cells Caco-2 and its possible mechanism.Methods Human colon cancer cells Caco-2 were induced with low-dose(20μg/L),medium-dose(60μg/L),and high-dose celastrus orbiculatus extract(180μg/L),30μmol/L cyclophosphamide(positive control)and blank control.In addition,Caco-2 cells either treated with blank control or high-dose celastrus orbiculatus extract were transfected with miR-NC,miR-378 mimics,anti-miR-NC and anti-miR-378.Nine replicates were set per group.Cell proliferation and apoptosis were examined by MTT assay and flow cytometry,respectively.Cell migration and invasion were detected by Transwell assay.were used to treat.Protein expressions of Ki67,matrix metalloproteinase-2(MMP-2),Cleaved-caspase-3,and MMP-9 were determined by Western blot.The mRNA level of miR-378 was detected by reverse transcription quantitative real-time polymerase chain reaction(qRT-PCR).Results Low-dose,medium-dose and high-dose Celastrus orbiculatus extract significantly decreased cell proliferation rate,protein levels of Ki67,MMP-2,and MMP-9,and number of migrating and invading cells(P<0.05),but significantly increased the expression of miR-378,apoptosis rate and protein level of Cleaved-caspase-3 in a dose-dependent manner(P<0.05).After transfection of miR-378 mimics,the cell proliferation rate,protein levels of Ki67,MMP-2 and MMP-9,and number of migrating and invading cells significantly decreased(P<0.05),and the apoptosis rate,the expression of Cleaved-caspase-3 and miR-378 significantly increased than those transfected with miR-NC(P<0.05).In comparison to those transfected with anti-miR-NC and induced with high-dose Celastrus orbiculatus extract,transfection of anti-miR-378 in cells induced with high-dose Celastrus orbiculatus extract significantly increased cell proliferation rate,protein levels of Ki67,MMP-2,and MMP-9,and number of migrating and invading cells(P<0.05),but significantly decreased the expression of miR-3

关 键 词：结肠肿瘤 南蛇藤 微小RNA-378 细胞增殖 肿瘤浸润 细胞凋亡 

分 类 号：R735.35[医药卫生—肿瘤] R285.5[医药卫生—临床医学]