甲状旁腺激素相关蛋白作为口腔鳞状细胞癌的预后指标。
Parathyroid hormone-related protein serves as a prognostic indicator in oral squamous cell carcinoma.
作者信息
Lv Zhongjing, Wu Xiangbing, Cao Wei, Shen ZongZe, Wang Lizhen, Xie FuRong, Zhang JianJun, Ji Tong, Yan Ming, Chen WanTao
机构信息
Department of Oral and Maxillofacial-Head and Neck Oncology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology, Shanghai, China.
出版信息
J Exp Clin Cancer Res. 2014 Dec 18;33(1):100. doi: 10.1186/s13046-014-0100-y.
BACKGROUND
In our previous study, parathyroid hormone-like hormone (PTHLH) which encodes parathyroid hormone-related protein (PTHrP) was revealed to be up-regulated in oral squamous cell carcinoma (OSCC) compared with paired apparently normal surgical margins using microarray method. However, the function and prognostic indicators of PTHLH/PTHrP in OSCC remain obscure.
METHODS
The mRNA levels of PTHLH and its protein levels were investigated in 9 OSCC cell lines and in 36 paired OSCC specimens by real-time PCR and western blotting. The biological function of PTHLH/PTHrP was investigated using small interfering RNA (siRNA) in 3 OSCC cell lines, and immunohistochemistry was used to estimate the prognostic value of PTHrP in 101 patients with head and neck squamous cell carcinoma (HNSCC), including OSCC and oropharyngeal squamous cell carcinoma. Cell cycle was tested by flow cytometry and cell cycle related genes were investigated by western blotting and immunocytochemistry assay.
RESULTS
This study showed that the mRNA and protein levels of PTHLH in 9 OSCC cell lines were much higher than that in normal epithelial cells (P < 0.0001). In 36 paired OSCC tissues, PTHLH mRNA expressions were found higher in 32 OSCC tissues than that of paired apparently normal surgical margins (P = 0.0001). The results revealed that the down-regulation of PTHLH/PTHrP by siRNAs could reduce cell proliferation and inhibit plate and soft agar colony formation as well as affect the cell cycle of OSCC cells. The key proteins related to the cell cycle were changed by anti-PTHLH siRNA. The results showed that cyclin D1 and CDK4 expressions were significantly reduced in the cells transfected with anti-PTHLH siRNA. On the other hand, the expression of p21 was increased. The results also showed that high PTHrP level was associated with poor pathologic differentiation (P = 0.0001) and poor prognosis (P = 0.0003) in patients with HNSCC.
CONCLUSIONS
This study suggests that PTHLH/PTHrP is up-regulated in OSCCs. Therefore, PTHLH/PTHrP could play a role in the pathogenesis of OSCC by affecting cell proliferation and cell cycle, and the protein levels of PTHrP might serve as a prognostic indicator for evaluating patients with HNSCCs.
背景
在我们之前的研究中，通过微阵列方法发现，编码甲状旁腺激素相关蛋白（PTHrP）的甲状旁腺激素样激素（PTHLH）在口腔鳞状细胞癌（OSCC）中相对于配对的明显正常手术切缘上调。然而，PTHLH/PTHrP在OSCC中的功能和预后指标仍不清楚。
方法
通过实时PCR和蛋白质印迹法，在9种OSCC细胞系和36对OSCC标本中研究PTHLH的mRNA水平及其蛋白质水平。在3种OSCC细胞系中使用小干扰RNA（siRNA）研究PTHLH/PTHrP的生物学功能，并且使用免疫组织化学评估PTHrP在101例头颈部鳞状细胞癌（HNSCC，包括OSCC和口咽鳞状细胞癌）患者中的预后价值。通过流式细胞术检测细胞周期，并且通过蛋白质印迹法和免疫细胞化学分析研究细胞周期相关基因。
结果
本研究表明，9种OSCC细胞系中PTHLH的mRNA和蛋白质水平远高于正常上皮细胞（P < 0.0001）。在36对OSCC组织中，发现32例OSCC组织中PTHLH mRNA表达高于配对的明显正常手术切缘（P = 0.0001）。结果显示，siRNA使PTHLH/PTHrP下调可减少细胞增殖并抑制平板和软琼脂集落形成，以及影响OSCC细胞的细胞周期。抗PTHLH siRNA改变了与细胞周期相关的关键蛋白。结果显示，用抗PTHLH siRNA转染的细胞中细胞周期蛋白D1和CDK4表达显著降低。另一方面，p21表达增加。结果还显示，HNSCC患者中高PTHrP水平与病理分化差（P = 0.0001）和预后差（P = 0.0003）相关。
结论
本研究提示PTHLH/PTHrP在OSCC中上调。因此，PTHLH/PTHrP可能通过影响细胞增殖和细胞周期在OSCC发病机制中起作用，并且PTHrP的蛋白质水平可能作为评估HNSCC患者的预后指标。
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