荷载LASP-1 siRNA的溶瘤腺病毒抑制人肾癌细胞增殖的研究  

Oncolytic adenovirus loaded with LASP-1 siRNA inhibits the proliferation of human renal carcinoma cells

作　　者：孙方浩 魏晋 曹希亮 张义静 高超 Sun Fanghao;Wei Jin;Cao Xiliang;Zhang Yijing;Gao Chao(Department of Urology,the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University,Xuzhou 221002,China)

机构地区：[1]徐州医科大学附属市立医院泌尿外科,徐州221002

出　　处：《国际泌尿系统杂志》2023年第3期466-470,共5页International Journal of Urology and Nephrology

基　　金：徐州医科大学附属医院科技发展基金项目(XYFM2020032);徐州医科大学附属市立医院2022年拔尖人才项目。

摘　　要：目的探讨荷载LASP-1 siRNA的溶瘤腺病毒(ZD55-LASP-1)对人肾癌细胞增殖的影响。方法选取ZD55-LASP-1、溶瘤腺病毒ZD55-EGFP感染人肾癌细胞(786-0细胞系),分别设为ZD55-LASP-1组和ZD55-EGFP组,以PBS作为阴性对照(阴性对照组)。采用Western blot检测ZD55-LASP-1感染细胞后的E1A和LASP-1蛋白表达情况,采用CCK8法检测ZD55-LASP-1对786-0细胞的增殖作用。采用细胞迁移和侵袭实验检测ZD55-LASP-1对786-0细胞迁移和侵袭的抑制作用。采用流式细胞术检测ZD55-LASP-1对786-0细胞凋亡的作用。结果ZD55-LASP-1和ZD55-EGFP感染后的786-0细胞有E1A蛋白的表达,而感染后的HK-2细胞无E1A蛋白的表达。病毒感染786-0细胞后,ZD55-LASP-1组和ZD55-EGFP组的LASP-1蛋白表达水平均低于阴性对照组(均P<0.05)。ZD55-LASP-1组的细胞存活率、穿膜的786-0细胞数、侵袭的细胞数均低于ZD55-EGFP组、阴性对照组(均P<0.05)。ZD55-LASP-1组的细胞凋亡率高于ZD55-EGFP组和阴性对照组(均P<0.05)。ZD55-LASP-1组的E-cadherin蛋白高于ZD55-EGFP组和阴性对照组,差异均有统计学意义(均P<0.05)。ZD55-LASP-1组的N-cadherin、Vimentin表达量低于ZD55-EGFP组和阴性对照组,差异均有统计学意义(均P<0.05)。结论ZD55-LASP-1可抑制人肾癌细胞的LASP-1基因表达,并诱导人肾癌细胞的凋亡作用。Objective To investigate the effect of oncolytic adenovirus(ZD55-LAP-1)loaded with LASP-1 siRNA on the proliferation of human renal cancer cells.Methods Human renal cancer cells(786-0 cell lines)infected with ZD55-LAP-1 and oncolytic adenovirus ZD55-EGFP were selected,and they were divided into ZD55-LAP-1 group and ZD55-EGFP group,respectively.PBS was used as a negative control(negative control group).Western blot was used to detect the effect of ZD55-LAP-1 on the expression of E1A and LASP-1 proteins in cells infected with ZD55-LAP-1,and CCK8 method was used to detect the effect of ZD55-LAP-1 on the proliferation of 786-0 cells.Cell migration and invasion experiments were conducted to detect the inhibitory effect of ZD55-LAP-1 on the migration and invasion of 786-0 cells.The effect of ZD55-LAP-1 on apoptosis of 786-0 cells was detected by flow cytometry.Results The 786-0 cells infected with ZD55-LAP-1 and ZD55-EGFP expressed E1A pro-tein,while the HK-2 cells infected with ZD55-EGFP did not express E1A protein.After virus infection of 786-0 cells,the expression level of LASP-1 protein in ZD55-LAP-1 and ZD55-EGFP groups was significantly lower than that in the negative control group(all P<0.05).The cell survival rate,the number of 786-0 cells penetrating the membrane,and the number of invasive cells in the ZD55-LAP-1 group were lower than those in the ZD55-EGFP group and the negative control group(all P<0.05).The apoptosis rate of ZD55-LAP-1 group was higher than that of ZD55-EGFP group and negative control group(all P<0.05).The E-cadherin protein in the ZD55-LAP-1 group was significantly higher than that in the ZD55-EGFP group and the negative control group,with significant differences(all P<0.05).The expression levels of N-cadherin and Vimentin in the ZD55-LAP-1 group were lower than those in the ZD55-EGFP group and the negative control group,with significant differences(all P<0.05).Conclusions ZD55-LAP-1 can inhibit the expression of LASP-1 gene in human renal cancer cells and induce apoptosis in human renal ca

关 键 词：肾肿瘤 溶瘤腺病毒 LASP-1 细胞增殖 

分 类 号：R737.11[医药卫生—肿瘤]