核素^125I标记hTERT/PSA启动子双调控溶瘤腺病毒对前列腺癌靶向治疗及肿瘤微环境的影响  

Experimental study of the effect of ^125I-RSOAds-hTERT/PSA oncolytic adenovirus on targeted therapy of prostate cancer and its effect on tumor microenvironment

作　　者：史振铎 魏振宁 郝林 庞昆 周家合 董秉政 张治国 赵岩 孙玉峰 韩从辉 Shi Zhenduo;Wei Zhenning;Hao Lin;Pang Kun;Zhou Jiahe;Dong Bingzheng;Zhang Zhiguo;Zhao Yan;Sun Yufeng;Han Conghui(Department of Urology Surgery,Xuzhou Central Hospital,Xuzhou Clinical School of Xuzhou Medical University,Xuzhou Central Hospital Affiliated to Medical School of Southeast University,Xuzhou 221009,China;School of Medicine,Jiangsu University,Zhenjiang 212013,China)

机构地区：[1]徐州市中心医院泌尿外科,徐州医科大学徐州临床学院,东南大学医学院附属徐州医院,221009 [2]江苏大学医学院,镇江212013

出　　处：《中华放射医学与防护杂志》2020年第8期573-581,共9页Chinese Journal of Radiological Medicine and Protection

基　　金：江苏省社会发展重点项目(BE2019637、BE2017635);江苏省医学创新团队(CXTD-2017048);江苏省青年医学人才(QNRC2016386);国家自然科学基金面上项目(81774089,81502387);江苏省高等学校自然科学研究面上项目(17KJB360001)。

摘　　要：目的探究125I-RSOAds-hTERT/PSA溶瘤腺病毒对前列腺癌靶向治疗作用以及对肿瘤微环境的影响。方法采用PCR扩增技术及双酶切连接技术构建125I-RSOAds-hTERT/PSA溶瘤腺病毒(125I-病毒复合物)。通过缺口末端标记法(TUNEL)染色,流式细胞实验以及Caspase-3的免疫印迹实验分别从体内和体外检测125I-病毒复合物对前列腺癌细胞的杀伤作用。通过酶联免疫吸附实验(ELISA)法检测人前列腺癌细胞株PC3和小鼠前列腺癌细胞株RM-1培养上清液及血清中的白介素2(IL-2)、IL-10、肿瘤坏死因子α(TNF-α)、γ干扰素(IFN-γ)等的分泌水平,探究125I-病毒复合物对瘤组织细胞因子分泌水平的影响。通过免疫组织化学法以及免疫荧光实验探究125I-病毒复合物对前列腺癌组织及癌细胞中CD24、CD44以及前列腺干细胞抗原(PSCA)表达的调节,同时检测瘤组织中CD32和血管内皮生长因子(VEGF)表达水平,以及CD4+、CD8+和巨噬细胞浸润情况。结果125I-病毒复合物体内、体外均可显著诱导癌细胞凋亡,同时显著高于125I组和病毒复合物组。同时IL-2(t=-183.30、-38.20,P<0.05)、IL-10(t=113.80、92.71,P<0.05)、TNF-α(t=-73.20、-73.91,P<0.05)、IFN-γ(t=-65.37、-139.70,P<0.05)在体内体外含量均升高。125I-病毒复合物可降低癌细胞及癌组织中CD24、CD44以及PSCA表达,减小癌组织重量(t=8.55,P<0.05),抑制癌组织血管生成,同时调节肿瘤组织中免疫反应。结论125I-病毒复合物溶瘤腺病毒对前列腺癌靶向可显著杀伤癌细胞,减少癌组织重量和血管生成,同时改善肿瘤微环境。Objective To investigate the effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on targeted therapy of prostate cancer and its effect on tumor microenvironment.Methods 125I-RSOAds-hTERT/PSA(125I-virus complex)oncolytic adenovirus was constructed by PCR amplification and double restriction enzyme ligation.TUNEL staining,flow cytometry and Caspase-3 immunoblotting assay were used to detect the killing effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on prostate cancer cells in vitro and in vivo,respectively.To explore the effect of 125I-virus complex on tumor tissue cytokine secretion levels,interleukin 2(IL-2),IL-10,tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)in the culture supernatant of human prostate cancer cell line PC3,mouse prostate adenocarcinoma cell line RM-1,and mice serum were detected by ELISA.We explored the regulation of 125I-virus complex on the expression of CD24,CD44 and prostate stem cell antigen(PSCA)in prostate tumor tissues and tumor cells through immunohistochemistry.Meanwhile,the expression levels of CD32 and vascular endothelial growth factor(VEGF),as well as CD4+,CD8+and macrophage infiltration in tumor tissue were detected through immunofluorescence experiments.Results 125I-virus complex oncolytic adenovirus significantly increased tumor cell apoptosis in vitro and in vivo that was significantly higher than that of 125I group and virus complex group.Meanwhile,IL-2(t=-183.30,-38.20,P<0.05),IL-10(t=113.80,92.71,P<0.05),TNF-α(t=-73.20,-73.91,P<0.05),IFN-γ(t=-65.37,-139.70,P<0.05)increased in vitro and in vivo.125I-virus complex reduced the expression of CD24,CD44 and PSCA in tumor cells and tumor tissue,reduced the weight of tumor tissue,inhibited angiogenesis of tumor tissue(t=8.55,P<0.05),and regulated the immune response in tumor tissue.Conclusions 125I-virus complex targeting prostate cancer can significantly kill cancer cells,reduce the weight and angiogenesis of tumor,and improve tumor microenvironment.

关 键 词：125I hTERT/PSA启动子双调控 溶瘤腺病毒 前列腺癌 靶向治疗 

分 类 号：R737.25[医药卫生—肿瘤]