摘要
目的:探讨普鲁卡因(PCA)及CXC趋化因子受体7(CXCR7)对膀胱癌细胞活力、迁移和侵袭的影响及潜在的作用机制。方法:用不同浓度PCA处理人膀胱癌RT4细胞,并将细胞分为PBS组(无PCA处理)、PCA组(4 mmol/L PCA处理)、siRNA阴性对照(si-Con)组(转染si-Con)、CXCR7 siRNA(si-CXCR7)组(转染si-CXCR7)、PCA+pcDNA组(4 mmol/L PCA处理并转染pcDNA)和PCA+pcDNA-CXCR7组(4 mmol/L PCA处理并转染pcDNACXCR7),siRNA和pcDNA均用脂质体法转染至RT4细胞。运用RT-qPCR检测RT4细胞中CXCR7的mRNA表达水平;采用CCK-8法检测各组细胞的活力;Transwell实验检测各组细胞的侵袭和迁移能力;Western blot检测各组细胞中CXCR7、AKT、STAT3、p-AKT和p-STAT3的蛋白水平。结果:相较于PBS组,不同浓度PCA处理后RT4细胞的活力、迁移能力和侵袭能力显著降低(P<0.05),PCA组RT4细胞中CXCR7的mRNA和蛋白表达水平显著降低(P<0.05)。相较于si-Con组,si-CXCR7组RT4细胞中CXCR7的mRNA和蛋白表达水平显著降低,且细胞的活力、迁移能力和侵袭能力显著降低(P<0.05);相较于PCA+pcDNA组,PCA+pcDNA-CXCR7组RT4细胞中CXCR7的mRNA和蛋白表达水平显著升高,且细胞的活力、迁移能力和侵袭能力显著提高(P<0.05)。相较于PBS组,PCA组的p-AKT和p-STAT3蛋白水平显著降低;相较于PCA+pcDNA组,PCA+pcDNA-CXCR7组的p-AKT和p-STAT3蛋白水平显著升高(P<0.05)。AKT和STAT3的蛋白水平在各组之间的差异无统计学显著性。结论:PCA可能通过抑制CXCR7的表达抑制膀胱癌细胞的活力、迁移和侵袭。过表达CXCR7可逆转PCA的作用,其机制可能与AKT/STAT3信号通路有关。
AIM:To investigate the effects of procaine(PCA)and CXC chemokine receptor 7(CXCR7)on the viability,migration and invasion of bladder cancer cells and its potential mechanism.METHODS:Human bladder cancer RT4 cells were treated with PCA at different concentrations,and were divided into PBS group(without PCA treatment),PCA group(treated with 4 mmol/L PCA),siRNA negative control(si-Con)group(transfected with si-Con),CXCR7 siRNA(si-CXCR7)group(transfected with si-CXCR7),PCA+pcDNA group(treated with 4 mmol/L PCA and transfected with pcDNA)and PCA+pcDNA-CXCR7 group(treated with 4 mmol/L PCA and transfected with pcDNACXCR7).The siRNA and pcDNA were transfected into the RT4 cells by liposome method.The mRNA expression of CXCR7 in the RT4 cells was detected by RT-qPCR.The cell viability was measured by CCK-8 assay.The invasion and migration abilities of the cells were detected by Transwell assays.The protein levels of CXCR7,AKT,STAT3,p-AKT and p-STAT3 were determined by Western blot.RESULTS:Compared with PBS group,the viability,migration ability and invasion ability of the RT4 cells treated with PCA at different concentrations were significantly decreased(P<0.05),and the expression of CXCR7 at mRNA and protein levels in PCA group was also significantly decreased(P<0.05).Compared with si-Con group,the expression of CXCR7 at mRNA and protein levels in si-CXCR7 group was significantly decreased,and the viability,migration ability and invasion ability of the cells were also significantly decreased(P<0.05).Compared with PCA+pcDNA group,the expression of CXCR7 at mRNA and protein levels in PCA+pcDNA-CXCR7 group was significantly increased,and the viability,migration ability and invasion ability of the cells were also significantly increased(P<0.05).Compared with PBS group,the protein levels of p-AKT and p-STAT3 in PCA group were significantly decreased(P<0.05).Compared with PCA+pcDNA group,the protein levels of p-AKT and p-STAT3 in PCA+pcDNACXCR7 group were significantly increased(P<0.05).No significant difference in the protein
作者
曾凯辉
吕慧莹
罗金泰
赵子良
ZENG Kai-hui;Lü Hui-ying;LUO Jin-tai;ZHAO Zi-liang(Department of Anesthesiology,The First Affiliated Hospital of Guangzhou Medical University,2Guangta Street Community Health Service Center,Yuexiu District,Guangzhou,3Department of Urology,The First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第4期612-618,共7页
Chinese Journal of Pathophysiology
基金
广东省医学科学技术基金资助项目(No.2015124141856933)。
