荷载LASP-1基因siRNA的溶瘤腺病毒对裸鼠人肾癌细胞移植瘤的抑制作用  

The effects of oncolytic adenovirus ZD55-LASP-1 on implanted human renal cancer in nude mice

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4期625-629,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基因siRNA的溶瘤腺病毒(ZD55-LASP-1)对裸鼠人肾癌786-0细胞移植瘤生长的抑制作用。方法建立人肾癌786-0细胞移植瘤模型,32只裸鼠成瘤成功后(肿瘤体积为100~150 mm^(3)),随机分为复制缺陷型腺病毒(Ad-LASP-1组)、溶瘤腺病毒(ZD55-EGFP组和ZD55-LASP-1组)、磷酸盐缓冲液(PBS组)四组,每组8只。定期测量肿瘤体积,采用免疫组化检测肿瘤组织内LASP-1蛋白的表达量;采用免疫荧光法检测移植瘤内腺病毒E1A基因的表达;采用原位末端标记法(TUNEL)检测肿瘤细胞凋亡。取裸鼠肝脏和肺脏组织进行HE染色了解有无肝脏损害和远处肺转移。结果ZD55-LASP-1组的裸鼠移植瘤的体积小于ZD55-EGFP组和Ad-LASP-1组,差异均有统计学意义(均P<0.05)。PBS组和Ad-LASP-1组无E1A蛋白表达,但ZD55-LASP-1组和ZD55-EGFP组可发现E1A表达。LASP-1蛋白免疫组化检测结果显示,与ZD55-EGFP组、Ad-LASP-1组和PBS组比较,ZD55-LASP-1组能够抑制靶基因LASP-1蛋白的表达,差异均有统计学意义(均P<0.05)。TUNEL实验结果显示,ZD55-LASP-1组的凋亡阳性率为(45.9±4.6)%,与Ad-LASP-1组、ZD55-EGFP组和PBS组[(26.3±3.1)%、(34.9±3.2)%、(13.8±1.6)%]比较,差异均有统计学意义(均P<0.05)。与Ad-LASP-1组和ZD55-EGFP组比较,ZD55-LASP-1可进一步促进肾癌细胞的凋亡。ZD55-LASP-1、ZD55-EGFP、Ad-LASP-1组均显示肿瘤组织生长抑制,表现为细胞核不同程度的深染,且核仁不明显。PBS组有3只裸鼠出现肿瘤肺转移,ZD55-EGFP组有1只出现肺转移,而ZD55-LASP-1组和Ad-LASP-1组未出现转移且未引起明显的肝组织变化。结论ZD55-LASP-1能有效抑制裸鼠人肾癌786-0细胞移植瘤的生长,为治疗肾癌提供新的平台。Objective To study the effects of oncolytic adenovirus expressing LASP-1 gene siRNA(ZD55-LASP-1)on the growth of implanted human renal cancer in nude mice.Methods The model of human renal carcinoma transplanted with 786-0 cells in nude mice was established.Thirty-two nude mice with tumor volume of 100-150 mm^(3) were randomly divided into 4 groups,with 8 mice in each group,including replication-defective adenovirus(Ad-LASP-1 group),oncolytic adenovirus(ZD55-EGFP group,ZD55-LASP-1 group)and phosphate buffer solution(PBS group).The volumeThe volume of tumor was measured regularly,and the expression of LASP-1 protein in tumor tissue was detected by immunohistochemistry.The expression of adenovirus E1A gene in transplanted tumor was detected by immunofluorescence method.Apoptosis of tumor cells was detected by in situ end labeling(TUNEL).HE staining was performed on liver and lung tissues of nude mice to investigate liver damage and distant lung metastasis.Results The volume of transplanted tumor in ZD55-LASP-1 group was lower than that in ZD55-EGFP group and Ad-LASP-1 group,and the difference was statistically significant(all P<0.05).There was no E1A protein expression in PBS group and Ad-LASP-1 group,but E1A expression was found in ZD55-LASP-1 group and ZD55-EGFP group.Immunohistochemical test results of LASP-1 protein showed that compared with ZD55-EGFP group,Ad-LASP-1 group and PBS group,ZD55-LASP-1 group could inhibit the expression of target gene LASP-1 protein(all P<0.05).TUNEL experimental results show that the The positive rate of apoptosis in ZD55-LASP-1 group was(45.9±4.6)%,and Ad-LASP-1group,ZD55-EGFP group and PBS group[(26.3±3.1)%、(34.9±3.2)%、(13.8±1.6)%],difference was statistically significant(all P<0.05).Compared with Ad-LASP-1 group and ZD55-EGFP group,ZD55-LASP-1 could further promote apoptosis of renal carcinoma cells.ZD55-LASP-1,ZD55-EGFP and Ad-LASP-1 groups all showed tumor tissue growth inhibition,which was manifested by different degrees of nuclear deep staining,and the nucleoli were

关 键 词：癌 肾细胞 Lasp-1基因 模型 动物 小鼠 

分 类 号：R737.11[医药卫生—肿瘤]