Radiosensitization of rat glioma with bromodeoxycytidine and adenovirus expressing herpes simplex virus-thymidine kinase delivered by slow, rate-controlled positive pressure infusion.
Radiosensitization of rat glioma with bromodeoxycytidine and adenovirus expressing herpes simplex virus-thymidine kinase delivered by slow, rate-controlled positive pressure infusion.
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通过缓慢、速率控制的正压输注，用溴脱氧胞苷和表达单纯疱疹病毒胸苷激酶的腺病毒对大鼠神经胶质瘤进行放射增敏。
DOI：
10.1038/sj.cgt.7700168
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Valerie,K
中科院分区：
文献类型：
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作者：
Brust,D;Feden,J;Farnsworth,J;Amir,C;Broaddus,WC;Valerie,K
用表达单纯疱疹病毒胸苷激酶(TK)的腺病毒(Adv-TK)感染体外培养的大鼠胶质瘤细胞，然后暴露于5-溴-2‘-脱氧胞苷(5-bromo-2’-deoxcytidine，BRDC)中，与AdβGal感染的细胞相比，5-溴-2‘-脱氧尿苷(BrdU)具有显著的放射增敏作用(增敏比为1.4-2.3)。细胞杀伤与BrdU DNA掺入增加和细胞凋亡密切相关。单纯照射(4Gy)诱导细胞凋亡效果相对较差，单纯单纯疱疹病毒TK/单纯树突状细胞组诱导的细胞凋亡呈剂量依赖性(10-100GyM)和时间依赖性(24-48h)增加，两者联合应用产生协同效应(1.5~2倍)。为了探讨ADV-TK/BRDC体内治疗的效果，在Fischer 344大鼠体内接种RT2细胞作为软组织肿瘤细胞，通过改变输液的体积和速度来优化病毒输注条件。我们发现，较大的病毒体积(100150μL)以≤1μL/分钟的速度递送是最优的，并且给出了均匀和可重复性的结果。使用这些最佳的输注条件，我们能够实现40%的腺病毒在肿瘤中的感染。用AdV-TK感染RT2肿瘤，并从渗透泵持续给药BRDC，结果显著(P<0.05)。001<；宝洁。009)与对照组相比，照射后6天肿瘤消退(30Gy2×5Gy3天)。用抗BrdU抗体对肿瘤切片进行原位染色，或对提取的肿瘤DNA和水解物进行高效液相分析，证实我们获得了高效、特异的BrdU掺入肿瘤细胞。这些结果表明，腺病毒介导的HSV-TK联合BRDC和放射治疗可能是治疗胶质瘤的一种有效的联合手段。
Infection of rat RT2 glioma cells in vitro with an adenovirus (ADV-TK) expressing herpes simplex virus (HSV) thymidine kinase (TK) and subsequent exposure to 5-bromo-2′-deoxycytidine (BrdC), which is specifically incorporated into ADV-TK-infected cell DNA as 5-bromo-2′-deoxyuridine (BrdU), results in significant radiosensitization (sensitizer enhancement ratio: 1.4–2.3) compared with Adβgal-infected cells. Cell killing correlated well with increased BrdU DNA incorporation and with apoptosis. Whereas radiation (4 Gy) alone was relatively ineffective in inducing apoptosis, treatment with HSV-TK/BrdC resulted in BrdC dose-(10–100 μM) and time-dependent (24–48 hours) increases, and the combination of the two treatments produced a synergistic response (1.5-to 2-fold). To investigate the effects of the ADV-TK/BrdC treatment in vivo, RT2 cells were grown as soft tissue tumors in Fischer 344 rats and conditions for virus infusion were optimized by altering the volume and rate of infusion using a rate-controlled positive pressure device. We found that relatively large volumes (100–150 μL) of virus delivered at rates of≤ 1 μL/minute were optimal and gave uniform and reproducible results. Using these optimal infusion conditions, we were able to achieve 40% adenovirus infection in the tumor. Infection of RT2 tumors with ADV-TK and continuous administration of BrdC from an osmotic pump resulted in significant (. 001< P<. 009) tumor regression 6 days after radiation (30 Gy delivered as 2× 5 Gy over 3 days) compared with controls. In situ staining of sectioned tumors with anti-BrdU antibody or by high-performance liquid chromatography analysis of extracted and hydrolyzed tumor DNA confirmed that we obtained efficient and specific incorporation of BrdU into tumor cells. These results suggest that adenovirus-mediated delivery of HSV-TK in combination with BrdC and radiation can potentially be an efficient combination modality for the treatment of gliomas.