Department of Ocular Pathology, Vision Research Foundation, Sankara Nethralaya, Tamil Nadu, India.
Invest Ophthalmol Vis Sci. 2011 Jul 23;52(8):5441-8. doi: 10.1167/iovs.10-6973.
To explore the possibility of stathmin as an effective therapeutic target and to evaluate the synergistic combination of stathmin RNAi and the antimicrotubule agents paclitaxel and vincristine to retinoblastoma Y79 cells.
RNAi-mediated specific inhibition of stathmin expression in Y79 cells was shown by real-time quantitative RT-PCR (RT-Q-PCR), its effect on cell proliferation by MTT assay, cell invasion using matrigel, microtubule polymerization by immunohistochemistry, apoptosis, cell cycle analysis by flow cytometry analysis, and the changes in FOXM1 protein expression were studied by Western blot. The effect of combination treatment of stathmin siRNA and paclitaxel/vincristine was studied by assessing cell viability and apoptosis.
Short interfering RNA-mediated transient stathmin downregulation resulted in a marked inhibition of retinoblastoma cell proliferation and cell invasion in vitro. Stathmin inhibition promoted Y79 cells to G2/M phase, and ultimately there were increased apoptotic events as evidenced by higher caspase-3 activation and cleaved poly(ADP-ribose) polymerase expression. Cells transfected with stathmin siRNA showed long and bundled microtubule polymers and sensitized the Y79 cells significantly to paclitaxel and vincristine.
Stathmin may be a pivotal determinant for retinoblastoma tumorigenesis and chemosensitivity. Strategies to inhibit stathmin will help to enhance the cytotoxic effect of paclitaxel while reducing toxicity (or side effects) to normal cells caused by high doses.
探索微管蛋白磷酸酶抑制剂 stathmin 作为一个有效的治疗靶点的可能性，并评估 stathmin RNAi 与抗微管药物紫杉醇和长春新碱联合应用对视网膜母细胞瘤 Y79 细胞的协同作用。
通过实时定量 RT-PCR（RT-Q-PCR）显示 Y79 细胞中 stathmin 的 RNAi 介导的特异性抑制，通过 MTT 测定法评估其对细胞增殖的影响，用基质胶进行细胞侵袭实验，用免疫组织化学检测微管聚合，通过流式细胞术分析进行细胞凋亡和细胞周期分析，并用 Western blot 检测 FOXM1 蛋白表达的变化。通过评估细胞活力和细胞凋亡来研究 stathmin siRNA 与紫杉醇/长春新碱联合治疗的效果。
短干扰 RNA 介导的 stathmin 瞬时下调导致体外视网膜母细胞瘤细胞增殖和侵袭明显抑制。Stathmin 抑制使 Y79 细胞进入 G2/M 期，最终导致更多的细胞凋亡事件，表现为更高的 caspase-3 激活和 cleaved poly(ADP-ribose) polymerase 表达。转染 stathmin siRNA 的细胞显示出长而束状的微管聚合物，并使 Y79 细胞对紫杉醇和长春新碱显著敏感。
Stathmin 可能是视网膜母细胞瘤发生和化疗敏感性的关键决定因素。抑制 Stathmin 的策略将有助于增强紫杉醇的细胞毒性作用，同时降低高剂量药物对正常细胞的毒性（或副作用）。
