麦冬皂苷D联合沉默环氧合酶-2基因对人胰腺癌BxPC-3细胞增殖、迁移和侵袭的影响  

Effects of Ophiopogon D combined with cyclooxygenase-2 silencing on proliferation,migration and invasion of human pancreatic cancer BxPC-3 cells

作　　者：钟扬[1,2] 何苗 刘志[1,2] 陈建宇[1,2] 张光年[1,2] 秦龙[3] 李婷[3] 李建水[1,2] Zhong Yang;He Miao;Liu Zhi;Chen Jianyu;Zhang Guangnian;Qin Long;Li Ting;Li Jianshui(Second Department of Hepatobiliary Surgery,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China;Institute of Hepato-Biliary-Pancreas and Intestinal Disease,North Sichuan Medical College,Nanchong 637000,China;Department of Gastrointestinal Surgery,Nanchong Central Hospital of Sichuan Province,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院肝胆外二科,南充637000 [2]川北医学院肝胆胰肠疾病研究所,南充637000 [3]四川省南充市中心医院胃肠外科,637000

出　　处：《国际肿瘤学杂志》2021年第10期583-590,共8页Journal of International Oncology

基　　金：南充市市校科技战略合作专项资金项目(18SXHZ0523)。

摘　　要：目的探讨麦冬皂苷D联合沉默环氧合酶-2(COX-2)基因对人胰腺癌BxPC-3细胞增殖、迁移、侵袭的影响。方法将BxPC-3细胞分为空白对照组、麦冬皂苷D高剂量组(40μmol/L)、中剂量组(20μmol/L)、低剂量组(10μmol/L);将沉默COX-2后的细胞分为空白组、COX-2抑制组(50 pmol/ml siRNA-COX-2)、麦冬皂苷D组(20μmol/L)与联合用药组(麦冬皂苷D 20μmol/L+50 pmol/ml siRNA-COX-2),采用CCK-8法检测细胞增殖活力,划痕实验检测细胞迁移距离,Transwell法检测细胞侵袭程度,采用实时定量PCR(RT-qPCR)法检测BxPC-3细胞中COX-2基因的相对表达水平,蛋白质印迹法检测BxPC-3细胞中COX-2、缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)蛋白相对表达水平。结果空白对照组,麦冬皂苷D高、中、低剂量组的细胞增殖率分别为(100.0±4.9)%、(71.8±5.4)%、(80.5±5.8)%和(89.7±5.7)%,细胞迁移距离分别为(279.8±24.0)μm、(141.9±21.2)μm、(168.8±37.1)μm和(224.6±19.9)μm,侵袭数量吸光度(A)值分别为1.107±0.095、0.390±0.030、0.596±0.017和0.826±0.034,差异均具有统计学意义(F=19.770,P<0.001;F=48.270,P<0.001;F=198.400,P<0.001),麦冬皂苷D高、中、低剂量组均明显低于空白对照组(均P<0.05);COX-2基因相对表达水平分别为1.007±0.178、0.387±0.169、0.567±0.142和0.740±0.030,蛋白相对表达水平分别为1.000±0.033、0.654±0.085、0.762±0.110和0.881±0.049,差异均具有统计学意义(F=10.280,P=0.004;F=11.780,P=0.003),麦冬皂苷D高、中剂量组均低于空白对照组(均P<0.05),麦冬皂苷D低剂量组与空白对照组差异均无统计学意义(均P>0.05);选择麦冬皂苷D中剂量(20μmol/L)作为后续实验浓度。沉默COX-2后,空白组、COX-2抑制组、麦冬皂苷D组和联合用药组的细胞增殖率分别为(100.0±2.8)%、(68.4±6.7)%、(67.7±5.9)%和(57.0±8.5)%,迁移距离分别为(274.4±23.8)μm、(217.0±18.8)μm、(186.2±18.6)μm和(115.7±15.8)μm,侵袭数量A值分别为1.143±0.092、Objective To explore the effects of Ophiopogon D combined with cyclooxygenase-2(COX-2)gene silencing on the proliferation,migration and invasion of human pancreatic cancer BxPC-3 cells.Methods BxPC-3 cells were divided into blank control group,Ophiopogonin D high-dose group(40μmol/L),medium-dose group(20μmol/L)and low-dose group(10μmol/L).The COX-2-slienced cells were divided into control group,COX-2 inhibited group(50 pmol/ml siRNA-COX-2),Ophiopogonin D group(20μmol/L)and combination treatment group(Ophiopogonin D 20μmol/L+50 pmol/ml siRNA-COX-2).The proliferation activity of BxPC-3 cells was detected by CCK-8,and the migration distance of BxPC-3 cells was detected by scratched assay.The invasion degree of BxPC-3 cells was detected by Transwell,the relative expression level of COX-2 gene in BxPC-3 cells was detected by real-time quantitative PCR(RT-qPCR),and the relative expressions of COX-2,hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)proteins in BxPC-3 cells were detected by Western blotting.Results The cell proliferation rates of blank control group,Ophiopogonin D high-dose,medium-dose and low-dose groups were(100.0±4.9)%,(71.8±5.4)%,(80.5±5.8)%and(89.7±5.7)%,respectively.The migration distances were(279.8±24.0)μm,(141.9±21.2)μm,(168.8±37.1)μm and(224.6±19.9)μm,respectively.The absorbance(A)values of invasion number were 1.107±0.095,0.390±0.030,0.596±0.017 and 0.826±0.034,respectively.There were statistically significant differences(F=19.770,P<0.001;F=48.270,P<0.001;F=198.400,P<0.001).The above indexes of the Ophiopogonin D high-,medium-and low-dose groups were significantly lower than those in the blank control group(all P<0.05).The relative expression levels of COX-2 gene were 1.007±0.178,0.387±0.169,0.567±0.142 and 0.740±0.030,respectively,and the relative protein expression levels were 1.000±0.033,0.654±0.085,0.762±0.110 and 0.881±0.049,respectively,with statistically significant differences(F=10.280,P=0.004;F=11.780,P=0.003).The above index

关 键 词：胰腺肿瘤 细胞增殖 肿瘤侵润 环氧化酶2 麦冬皂苷D 

分 类 号：R735.9[医药卫生—肿瘤]