靶向 HSP90/Survivin 使用基于细胞通透结构的肽模拟索拉非尼在视网膜母细胞瘤中的应用。

Targeting HSP90/Survivin using a cell permeable structure based peptido-mimetic shepherdin in retinoblastoma.

机构信息

Department of Larsen & Toubro Ocular Pathology, Vision Research Foundation, Sankara Nethralaya, 18/41, College Road, Chennai 600006, India; Birla Institute of Technology and Science (BITS), Pilani, Rajasthan 333031, India.

Nanomedicine-Laboratory of Immunology and Molecular Biomedical Research (NLIMBR), School of Medicine (SoM), Molecular and Medical Research (MMR) Strategic Research Centre, Faculty of Health, Deakin University, Pigdons Road, Waurn Ponds, Geelong, Victoria 3217, Australia.

出版信息

Chem Biol Interact. 2016 May 25;252:141-9. doi: 10.1016/j.cbi.2016.04.011. Epub 2016 Apr 7.

DOI:10.1016/j.cbi.2016.04.011

PMID:27062892

Abstract

BACKGROUND

Retinoblastoma (RB) is a childhood retinal malignancy. Effective therapeutic strategies are still being investigated in RB disease management. Here, the anti-cancer effect of shepherdin, a peptido-mimetic inhibiting heat shock protein (HSP90)-Survivin interaction has been analyzed.

METHODS

We analyzed HSP (HSP70/90) and Survivin protein expressions by immunohistochemistry (29 archival tumors), qRT-PCR, FACS and Western analysis (10 un-fixed RB tumors). We also analyzed cellular cytotoxicity and anti-proliferative effect in peptide treated RB cells (Y79, Weri Rb1) and MIO-M1 cells.

RESULTS

Heterogeneous expressions of HSP70/90 and Survivin with a significant association between HSP70 and HSP90 (r(2) = 0.59, p = 0.001) was observed. In RB cells, anti-tumor effects were detected with 0.42 μg/ml of shepherdin at 4 h s of serum starvation. Decreased Survivin, Bcl2, MMP-2 activity with increased Bax, Bim, and Caspase-9 protein expressions were noticed. No significant changes were observed in shepherdin treated non-neoplastic MIO-M1, nor in scramble-peptide treated RB cells.

CONCLUSION

The presence of HSPs (HSP70/90) and Survivin reveals multiple cellular mechanisms adopted by RB cells during cancer progression. Serum starvation induced HSP90 whose interactions with Survivin were specifically inhibited by shepherdin. The associated molecular shuffling has been reported. These findings strongly implicate the potential of targeting HSP90-Survivin interaction as an adjuvant therapy in RB management.

摘要

背景

视网膜母细胞瘤（RB）是一种儿童视网膜恶性肿瘤。在 RB 疾病管理中，仍在探索有效的治疗策略。在这里，分析了一种抑制热休克蛋白（HSP90）-Survivin 相互作用的肽模拟物——shepherdin 的抗癌作用。

方法

我们通过免疫组织化学（29 个存档肿瘤）、qRT-PCR、FACS 和 Western 分析分析了 HSP（HSP70/90）和 Survivin 蛋白的表达（10 个未固定的 RB 肿瘤）。我们还分析了肽处理的 RB 细胞（Y79、Weri Rb1）和 MIO-M1 细胞中的细胞毒性和抗增殖作用。

结果

观察到 HSP70/90 和 Survivin 的异质性表达，HSP70 和 HSP90 之间存在显著相关性（r²=0.59，p=0.001）。在 RB 细胞中，在血清饥饿 4 小时时，用 0.42μg/ml 的 shepherdin 检测到抗肿瘤作用。发现 Survivin、Bcl2、MMP-2 活性降低，Bax、Bim、Caspase-9 蛋白表达增加。在非肿瘤性 MIO-M1 的 shepherdin 处理或 scramble 肽处理的 RB 细胞中未观察到明显变化。

结论

HSPs（HSP70/90）和 Survivin 的存在揭示了 RB 细胞在癌症进展过程中采用的多种细胞机制。血清饥饿诱导 HSP90，其与 Survivin 的相互作用被 shepherdin 特异性抑制。已报道相关的分子洗牌。这些发现强烈暗示了靶向 HSP90-Survivin 相互作用作为 RB 管理辅助治疗的潜力。