Department of Ophthalmology, Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, PR China.
Br J Ophthalmol. 2012 Oct;96(10):1331-8. doi: 10.1136/bjophthalmol-2011-301470. Epub 2012 Jul 27.
Deregulation of Bcl2 pathway is implicated in the pathogenesis of uveal melanoma (UM). Oncolytic adenovirus H101 is the world's first oncolytic viral therapy for cancer approved for clinical use. We aimed to explore a potential synergy of downregulating Bcl2 pathway using a small interfering RNA (siBCL2) combined with H101 therapy on UM cell lines.
The sensitivity to adenovirus infection was analysed by flow cytometry. PCR, real-time-PCR and western blot were used to detect Bcl2, p53, Bax and fibre expression. Appropriate multiplicity of H101 infection and cell survival rate were measured by a cell counting kit-8 assay. UM cells were stained with Annexin-V and propidium iodide for apoptosis assay and cell cycle distribution.
VUP cells (without elevation of Bcl2) exhibited greater sensitivity to adenovirus infection than OM431 cells (Bcl2 elevated cell line). Bcl2 expression was markedly reduced by siBCL2 or siBCL2 plus H101. Combined treatment with siBCL2 and H101 produced substantial growth inhibition of OM431 cells by enhancing apoptosis and cell cycle arrest through Bax-p53-induced apoptotic pathway.
SiBCL2 and H101 exhibited synergistic cytotoxic effect in Bcl2 elevated UM cell lines and could potentially serve as a novel targeted molecular therapy for UM.
Bcl2 通路的失调与葡萄膜黑色素瘤（UM）的发病机制有关。溶瘤腺病毒 H101 是世界上第一个获准临床使用的溶瘤病毒癌症治疗药物。我们旨在探索使用小干扰 RNA（siBCL2）联合 H101 治疗下调 Bcl2 通路的潜在协同作用，用于 UM 细胞系。
通过流式细胞术分析对腺病毒感染的敏感性。PCR、实时 PCR 和 Western blot 用于检测 Bcl2、p53、Bax 和纤维表达。通过细胞计数试剂盒-8 测定适当的 H101 感染倍数和细胞存活率。用 Annexin-V 和碘化丙啶对 UM 细胞进行染色，进行凋亡检测和细胞周期分布。
VUP 细胞（Bcl2 未升高）对腺病毒感染的敏感性高于 OM431 细胞（Bcl2 升高细胞系）。siBCL2 或 siBCL2 加 H101 显著降低了 Bcl2 的表达。siBCL2 和 H101 的联合治疗通过 Bax-p53 诱导的凋亡途径增强凋亡和细胞周期停滞，对 OM431 细胞产生了显著的生长抑制作用。
siBCL2 和 H101 在 Bcl2 升高的 UM 细胞系中表现出协同细胞毒性作用，可能作为 UM 的一种新型靶向分子治疗方法。
