溶瘤腺病毒ZD55．特异性核基质结合区结合蛋白1对雄激素非依赖前列腺癌细胞DU145增殖和侵袭的影响  

Inhibitory effect of oncolytic adenovirus ZD55 - special AT rich sequence binding protein 1 on pro- liferation and invasion of andragon - independent prostate cancer DU145 cells

作　　者：仇存权 陈家存[1] 

机构地区：[1]徐州医学院附属医院泌尿外科,江苏221002

出　　处：《中华实验外科杂志》2017年第1期82-85,共4页Chinese Journal of Experimental Surgery

基　　金：江苏省自然科学基金（BK20151166）

摘　　要：目的观察携带特异性核基质结合区结合蛋白1（SATB1）基因短发卡RNA（shRNA）的溶瘤腺病毒ZD55-SATB1对雄激素非依赖前列腺癌细胞DU145增殖和侵袭的影响。 方法 同源重组法包装溶瘤腺病毒ZD55-SATB1；转染倍数（MOI）＝10.0的病毒感染DU145细胞72 h后，反转录-聚合酶链反应（RT-PCR）法检测ZD55-SATB1对前列腺癌细胞SATB1基因的mRNA表达的影响；Western blot检测ZD55-SATB1对前列腺癌细胞SATB1蛋白表达的影响及病毒复制能力；结晶紫染色法、细胞计数试剂盒（CCK-8）法检测ZD55-SATB1对细胞的增殖抑制和毒性作用；细胞侵袭试验检测细胞侵袭力改变。结果 成功包装病毒ZD55-SATB1；RT-PCR和Western blot结果显示：ZD55-SATB1组SATB1表达量明显减少，与对照组比较差异有统计学意义。E1A蛋白表达水平在前列腺癌细胞DU145显著高于人正常前列腺上皮细胞PZ-HPV-7，差异有统计学意义，表明ZD55-SATB1具有肿瘤靶向性；结晶紫染色示：ZD55-SATB1组DU145有明显的细胞病效应。CCK-8结果显示：细胞感染后4 d，ZD55-SATB1组细胞存活率为（31.60±3.31）%，显著低于ZD55-增强型绿色荧光蛋白（EGFP）组（61.70±2.42）%（P＝ 0.025）和Ad-SATB1组（82.40±3.54）%（P＝ 0.017），差异有统计学意义。Transwell细胞侵袭实验结果显示：ZD55-SATB1组侵袭细胞数为（61.30±17.25）个，显著低于Ad-SATB1组[（170.10±25.44）个，P＝ 0.043]，ZD55-EGFP组[（295.40±23.36）个，P＝0.013]和磷酸盐缓冲液（PBS）组[（490.50±33.36）个，P＝0.009]，与对照组比较差异有统计学意义。结论 携带SATB1基因shRNA的溶瘤腺病毒ZD55-SATB1可有效沉默前列腺癌细胞DU145中SATB1基因的表达，同时有效抑制DU145细胞的增殖和侵袭。Objective To investigate the suppressive effect of oncolytic adenovirus ZD55-special AT rich sequence binding protein 1 （SATB1） expressing small hairpin RNA targeting SATB1 gene on growth and invasion of prostate cancer DU145 cells.Methods The oncolytic adenovirus ZD55-SATB1 was constructed and identifies. The inhibitory effect of ZD55-SATB1 on SATB1 expression was evaluated by reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting analysis at 72 h after infection of DU145 cells with ZD55-SATB1 at a MOI of 10. The cytotoxicity of ZD55-SATB1 was detected by cell counting kit-8 （CCK-8） assay and crystal violet staining assay. Cell invasion was detected by Matrigel invasion assay. The expression of E1A protein in DU145 cell lines infected with viruses respectively was detected by Western blotting.Results The oncolytic adenoviruses ZD55-SATB1 were successfully constructed and identified. RT-PCR and Western blotting assay showed that ZD55-SATB1 could significantly decrease the expression of SATB1 in DU145 cells （P〈0.05）. The relative protein levels of E1A in DU145 cells treated with ZD55-SATB1 were significantly higher than those in human normal prostate epithelial cell line PZ-HPV-7 cells （P〈0.01）, which indicated cancer specificity of ZD55-SATB1 for prostate cance cells. Crystal violet staining assay showed that ZD55-SATB1 exhibited obvious cytotoxicity in DU145 cells. The cytopathic effect of ZD55-SATB1 was much stronger than that of ZD55-enhanced green fluorescent protein （EGFP） and Ad-SATB1. CCK-8 assay showed that cell viability of ZD55-SATB1 group [（31.60±3.31）%] was significantly lower than that in ZD55-EGFP group [（61.70±2.42）%, P=0.025] and Ad-SATB1 group [（82.40±3.54）%, P=0.017] for DU145 cells after 4 days. Transwell cell invasive experiment showed that the number of cells that invaded the membrane in ZD55-SATB1 group was （61.30±17.25） cells, significantly less than that in the Ad-SATB1 group [（170.10±25.44） cells, P=0.043], the ZD

关 键 词：溶瘤腺病毒 特异性核基质结合区结合蛋白1 增殖 侵袭 

分 类 号：R737.25[医药卫生—肿瘤]