紫花牡荆素通过抑制7次跨膜超蛋白家族成员4的表达抑制膀胱癌细胞增殖迁移和侵袭  

Casticin inhibits the proliferation,migration and invasion of bladder cancer cells by inhibition of TM7SF4 expression

作　　者：徐豪[1] 石红林[1] 郝建伟[1] 束坤鹏 张云天 侯铁奇 Xu Hao;Shi Honglin;Hao Jianwei;Shu Kunpeng;Zhang Yuntian;Hou Tieqi(Department of Urology,Henan People′s Hospital,Zhengzhou 450000,China;Medical College of Henan University,Kaifeng 475004,China)

机构地区：[1]河南省人民医院泌尿外科,郑州450000 [2]河南大学医学院,开封475004

出　　处：《中华肿瘤杂志》2022年第4期334-340,共7页Chinese Journal of Oncology

摘　　要：目的探讨紫花牡荆素(CAS)对膀胱癌T24细胞增殖、迁移和侵袭的影响及作用机制。方法体外培养T24细胞, 分为对照组、5、10、20 μmol/L CAS组、si-NC组、si-TM7SF4组、CAS+pcDNA组和CAS+pcDNA-TM7SF4组, 采用细胞计数试剂盒8(CCK-8)法检测各组细胞的增殖抑制情况, Transwell法检测细胞迁移和侵袭的能力, Western blot法检测细胞中细胞周期蛋白D1(cyclin D1)、p21、基质金属蛋白酶2(MMP-2)、MMP-9和7次跨膜超蛋白家族成员4(TM7SF4)蛋白的表达水平, 实时荧光定量PCR检测TM7SF4 mRNA的表达水平。结果 5、10、20 μmol/L CAS组T24细胞增殖抑制率分别为(17.68±1.41)%、(33.54±3.16)%和(61.44±5.50)%, 均高于对照组[(0.00±0.00)%, 均P<0.001]。5、10、20 μmol/L CAS组细胞迁移数分别为(72.83±5.66)个、(59.13±4.27)个和(41.25±3.22)个, 细胞侵袭数分别为(55.83±5.15)个、(42.19±3.06)个和(31.13±3.22)个, 均低于对照组[分别为(86.11±5.16)个和(68.82±5.29)个, 均P<0.001]。5、10、20 μmol/L CAS组T24细胞中cyclin D1、MMP-2、MMP-9、TM7SF4蛋白表达水平、TM7SF4 mRNA表达水平均低于对照组(均P<0.001), p21蛋白表达水平高于对照组(P<0.001)。si-TM7SF4组T24细胞增殖抑制率[(50.35±4.67)%]高于si-NC组[(6.31±0.58)%, P<0.001], si-TM7SF4组细胞迁移数[(53.51±4.18)个]和细胞侵袭数[(42.92±3.81)个]均低于si-NC组[分别为(85.26±4.99)个和(67.93±4.64)个, 均P<0.001]。si-TM7SF4组T24细胞中TM7SF4、cyclinD1、MMP-2和MMP-9蛋白表达水平均低于si-NC组(均P<0.001), p21蛋白表达水平高于si-NC组(P<0.001)。CAS+pcDNA-TM7SF4组T24细胞增殖抑制率[(21.45±2.46)%]低于CAS+pcDNA组[(64.06±4.49)%, P<0.001], CAS+pcDNA-TM7SF4组细胞迁移数[(75.66±6.57)个]和侵袭数[(59.35±5.40)个]均高于CAS+pcDNA组[分别为(40.43±3.85)个和(30.25±3.32)个, 均P<0.001]。CAS+pcDNA-TM7SF4组T24细胞中TM7SF4、cyclinD1、MMP-2和MMP-9蛋白表达水平均高于CAS+pcDNA组(均P<0.001), p21蛋白表达水平低于CAS+pcDObjective To explore the effect and mechanism of Casticin(CAS)on the proliferation,migration and invasion of bladder cancer T24 cells.Methods T24 cells were cultured in vitro and divided into control group,5,10,20μmol/L CAS groups,si-NC group,si-TM7SF4 group,CAS+pcDNA group and CAS+pcDNA-TM7SF4 group.Cell counting kit-8(CCK-8)was used to detect cell proliferation;Transwell was used to detect cell migration and invasion;western blot was used to detect the protein expressions of cyclin D1,p21,MMP-2,MMP-9 and TM7SF4,and real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used to detect the expression of TM7SF4 mRNA.Results The inhibition rates of T24 cells in the 5,10,20μmol/L CAS groups were(17.68±1.41)%,(33.54±3.16)%and(61.44±5.50)%,respectively,higher than(0.00±0.00)%of the control group(P<0.001),but the numbers of migration and invasion were 72.83±5.66,59.13±4.27,41.25±3.22 and 55.83±5.15,42.19±3.06,31.13±3.22,respectively,lower than 86.11±5.16 and 68.82±5.29 of the control group(P<0.001).The protein expression levels of cyclin D1,MMP-2,MMP-9,TM7SF4 and the expression levels of TM7SF4 mRNA in the 5,10,and 20μmol/L CAS groups were lower than the control group(P<0.001).However,the protein expression levels of p21 were 0.37±0.03,0.51±0.04,and 0.66±0.06,respectively,higher than 0.25±0.03 in the control group(P<0.001).The inhibition rate of T24 cells in the si-TM7SF4 group was(50.35±4.67)%,higher than(6.31±0.58)%in the si-NC group(P<0.001),but the numbers of migration and invasion were 53.51±4.18 and 42.92±3.81,lower than 85.26±4.99 and 67.93±4.64 of the si-NC group(P<0.001).The protein expression levels of TM7SF4,CyclinD1,MMP-2,MMP-9 in the si-TM7SF4 group were lower than the si-NC group(P<0.001).However,the protein expression level of p21 in the si-TM7SF4 group was higher than the si-NC group(P<0.001).The inhibitory rate of T24 cells in the CAS+pcDNA-TM7SF4 group was(21.45±2.46)%,lower than(64.06±4.49)%of the CAS+pcDNA group(P<0.001),but the number of migrat

关 键 词：膀胱肿瘤 紫花牡荆素 7次跨膜超蛋白家族成员4 增殖 迁移 侵袭 

分 类 号：R285[医药卫生—中药学]