Morris B D, Drazan K E, Csete M E, Werthman P E, Van Bree M P, Rosenthal J T, Shaked A
Department of Surgery, University of California at Los Angeles 90024-6904.
J Urol. 1994 Aug;152(2 Pt 1):506-9. doi: 10.1016/s0022-5347(17)32783-0.
This study was designed to examine the potential for gene therapy in bladder in vivo using adenoviral vectors. Gene transfer to rat bladders was accomplished via direct intravesical instillation using a replication-defective adenoviral vector containing a marker gene encoding for Escherichia coli beta-galactosidase (beta-gal). Successful gene transfer was confirmed by analyzing bladder samples for DNA and RNA using polymerase chain reaction (PCR) with primers specific for beta-gal and adeno sequences, detecting beta-gal in full-thickness bladder wall using specific histochemical staining (X-gal) and documenting recombinant protein production. Bladder architecture was preserved, without evidence of distant spread of virus as assessed by PCR. Gene expression was evident for at least 7 days. In summary, bladder cells can be genetically altered using replication-deficient adenoviral vectors via simple intravesical instillation of vector. Introduction of exogenous genetic material is a potentially powerful therapeutic modality for immunomodulation of bladder neoplasms.
本研究旨在探讨使用腺病毒载体在体内对膀胱进行基因治疗的潜力。通过使用含有编码大肠杆菌β-半乳糖苷酶（β-gal）的标记基因的复制缺陷型腺病毒载体，经膀胱内直接滴注实现向大鼠膀胱的基因转移。通过使用针对β-gal和腺病毒序列的特异性引物，采用聚合酶链反应（PCR）分析膀胱样本中的DNA和RNA，使用特异性组织化学染色（X-gal）检测全层膀胱壁中的β-gal，并记录重组蛋白的产生，证实了成功的基因转移。膀胱结构得以保留，通过PCR评估未发现病毒远处扩散的证据。基因表达至少持续7天。总之，通过简单的膀胱内滴注载体，使用复制缺陷型腺病毒载体可对膀胱细胞进行基因改造。引入外源性遗传物质是一种对膀胱肿瘤进行免疫调节的潜在有力治疗方式。
