Fu Y-R, Chen F, Luo Y, Yi Y-F
Department of Urology, The Peoples Hospital of Nanchuan Chongqing, Chongqing, China.
Eur Rev Med Pharmacol Sci. 2019 Jan;23(1):75-86. doi: 10.26355/eurrev_201901_16751.
Bladder cancer is the 2nd most common reason for human genitourinary cancer-associated mortality. This study aimed to investigate the effects of Nanoscale bubble ultrasound contrast agents-mediated yeast-cytosine-deaminase-thymidine kinase/ganciclovir (YCD-TK/GCV) or YCD-TK/5-fluorocytosine (5-FC) suicide gene therapy system on BIU-87 cell growth.
Targeted nanoscale bubble ultrasound contrast agents were prepared by utilizing thin-film hydration-sonication approach. Nanoscale bubble-LV5-YCD-TK/GCV(5-FC) was constructed and transfected to BIU-87 cells. Hematoxylin and eosin (HE) staining was used to evaluate inflammation. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to examine cell viability. Cell-cycle distribution was analyzed with cell cycle assay. Flow cytometry assay was utilized to test apoptosis of BIU-87 cells. YCD-TK expression was examined using Western blot and quantitative Real Time-PCR (qRT-PCR), respectively.
YCD-TK highly expressed in Nanoscale bubble mediated suicide gene therapy system. Nanoscale bubble-mediated suicide gene therapy system significantly induced inflammatory response and apoptosis compared to that of Nanoscale bubble group (p<0.05). Nanoscale bubble mediated suicide gene therapy system significantly reduced cell viability compared to that of the Nanoscale bubble group (p<0.05). Nanoscale bubble mediated suicide gene therapy system significantly inhibited cell cycle arrest compared to that of the Nanoscale bubble group (p<0.05). Nanoscale bubble-LV5-YCD-TK/GCV/5-FC therapy system significantly reduced BIU-87 cell viability compared to that of the Nanoscale bubble-associated groups (p<0.05).
Nanoscale bubble-mediated suicide gene therapy system, bubble-LV5-YCD-TK/GCV/5-FC, acts as a novel therapeutic strategy for bladder cancer treatment.
膀胱癌是人类泌尿生殖系统癌症相关死亡的第二大常见原因。本研究旨在探讨纳米级气泡超声造影剂介导的酵母胞嘧啶脱氨酶-胸苷激酶/更昔洛韦（YCD-TK/GCV）或YCD-TK/5-氟胞嘧啶（5-FC）自杀基因治疗系统对BIU-87细胞生长的影响。
采用薄膜水化-超声法制备靶向纳米级气泡超声造影剂。构建纳米级气泡-LV5-YCD-TK/GCV（5-FC）并转染至BIU-87细胞。采用苏木精和伊红（HE）染色评估炎症。采用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐（MTT）法检测细胞活力。用细胞周期分析法分析细胞周期分布。采用流式细胞术检测BIU-87细胞凋亡。分别用蛋白质免疫印迹法和实时定量聚合酶链反应（qRT-PCR）检测YCD-TK的表达。
YCD-TK在纳米级气泡介导的自杀基因治疗系统中高表达。与纳米级气泡组相比，纳米级气泡介导的自杀基因治疗系统显著诱导炎症反应和细胞凋亡（p<0.05）。与纳米级气泡组相比，纳米级气泡介导的自杀基因治疗系统显著降低细胞活力（p<0.05）。与纳米级气泡组相比，纳米级气泡介导的自杀基因治疗系统显著抑制细胞周期停滞（p<0.05）。与纳米级气泡相关组相比，纳米级气泡-LV5-YCD-TK/GCV/5-FC治疗系统显著降低BIU-87细胞活力（p<0.05）。
纳米级气泡介导的自杀基因治疗系统，即气泡-LV5-YCD-TK/GCV/5-FC，是一种治疗膀胱癌的新型治疗策略。
