Department of Oral & Maxillofacial-Head & Neck Oncology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, No 639, Zhizaoju Rd, Shanghai, 200011, China.
Department of Oral Pathology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
J Exp Clin Cancer Res. 2017 Aug 14;36(1):109. doi: 10.1186/s13046-017-0575-4.
The aim of this study was to investigate the oncogenic function and regulatory mechanism of stathmin in oral squamous cell carcinoma (OSCC).
Two-dimensional electrophoresis and liquid chromatography-tandem mass chromatography were applied to screen differentiated proteins during carcinogenesis in OSCC. Cell Counting Kit-8 (CCK-8) assays, colony formation, migration, flow cytometry, immunofluorescence and a xenograft model were used to detect the function of stathmin. The correlation between stathmin and p53 expression was analyzed using immunohistochemistry. Mutant/wild type p53 plasmids and small interfering RNA were used to examine the regulation of stathmin. Chromatin immunoprecipitation assays and luciferase assays were performed to detect the transcriptional activation of stathmin by p53.
Overexpression of stathmin was screened and confirmed in OSCC patients and cell lines. Silencing expression of stathmin inhibited proliferation, colony formation and migration and promoted apoptosis. Poly ADP ribose polymerase (PARP) and cyclin-dependent kinase 1 (cdc2) were activated after silencing the expression of stathmin. Suppression of tumorigenicity was also confirmed in vivo. Mutant p53 transcriptionally activated the expression of stathmin in HN6 and HN13 cancer cells, but not in HN30 cells harboring wild type p53.
These results suggest that stathmin acts as an oncogene and is transcriptionally regulated by mutant p53, but not by wild-type p53. Stathmin could be a potential anti-tumor therapeutic target in OSCC.
本研究旨在探讨原癌基因 stathmin 在口腔鳞状细胞癌（OSCC）中的致癌功能和调控机制。
应用二维电泳和液相色谱-串联质谱技术筛选 OSCC 癌变过程中的差异表达蛋白。采用细胞计数试剂盒（CCK-8）检测、集落形成实验、迁移实验、流式细胞术、免疫荧光和异种移植模型来检测 stathmin 的功能。采用免疫组织化学分析 stathmin 与 p53 表达的相关性。利用突变型/野生型 p53 质粒和小干扰 RNA 来研究 stathmin 的调控作用。采用染色质免疫沉淀实验和荧光素酶实验来检测 p53 对 stathmin 的转录激活作用。
在 OSCC 患者和细胞系中筛选并证实了 stathmin 的过表达。沉默 stathmin 的表达抑制了增殖、集落形成和迁移，并促进了细胞凋亡。沉默 stathmin 的表达后，多聚 ADP 核糖聚合酶（PARP）和细胞周期蛋白依赖性激酶 1（cdc2）被激活。体内也证实了抑制肿瘤生成的作用。突变型 p53 在 HN6 和 HN13 癌细胞中转录激活了 stathmin 的表达，但在携带野生型 p53 的 HN30 细胞中则没有。
这些结果表明，stathmin 作为一种癌基因，其转录受到突变型 p53 的调控，而不受野生型 p53 的调控。stathmin 可能成为 OSCC 潜在的抗肿瘤治疗靶点。
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