Biomarkers in nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) is one of the most common malignancies in certain areas of Southern China, Southeastern Asia and Northern Africa. Currently, evaluation of NPC prognosis is mainly based on the tumor-node-metastasis (TNM) staging system. However, NPC patients with the same clinical stage often present different clinical courses, suggesting that the TNM staging is insufficient to predict prognosis of this disease. Therefore, it is important to find molecular biomarkers, which can help clinicians to identify NPC patients with worse prognosis and develop therapeutic interventions in NPC patients. This thesis presents the identification and investigation of mechanism of several novel markers in NPC. In the first paper, Caveolin-1 (Cav-1), a major structural component of caveolae, and CD147 (also known as extracellular matrix metalloproteinase inducer, EMMPRIN), a glycoprotein, were found to be overexpressed in NPC. Both Cav-1 and CD147 expression levels correlated significantly with metastasis and poor prognosis of NPC patients. Further studies revealed that Cav-1 and CD147 enhance NPC cell migration, which is associated with MMP-3 and MMP-11 (active) secretion. The role of microRNA-155 (miR-155) is associated with oncogenesis of several human tumors. In the second paper, miR-155 was found to be upregulated in NPC cell lines and clinical samples. EBV encoded LMP1 and LMP2A could further enhance the expression of miR-155 in NPC CNE1 and TW03 cells. JMJD1A was identified as a direct target of miR-155 in NPC. Downregulation of JMJD1A was significantly correlated with N stage of the TNM classification, a lower five-year survival rate, and a lower five-year disease-free survival rate of NPC patients. Spleen tyrosine kinase (Syk) is a nonreceptor tyrosine kinase and often aberrantly expressed in human cancers. In the third paper, high expression of Syk was detected in 24% of NPC cases. High expression of Syk, resulted partly from LMP2A expression in NPC, is associated with tumor recurrence and poor prognosis of NPC patients. Human chromosome 3 (Chr. 3) contains clusters of tumor suppressor genes (TSGs) involved in many cancer types. In the fourth paper, using Not I Chr. 3 microarray, ten candidate TSGs were found in NPC. Among them, the CpG island in the promoter region of Wingless-type Mouse mammary tumor virus integration site family, member 7A (WNT7a) and the intron 1 region of Integrin α9 (ITGA9) were confirmed to be hypermethylated in NPC by bisulfite sequencing and methylation specific PCR. Demethylating agent 5-aza-2′-deoxycytidine (5-aza-CdR) treatment could restore the expression of WNT7a and ITGA9 in NPC cell lines. Furthermore, WNT7a and ITGA9 were downregulated in NPC clinical samples. As both proteins execute significant functions related to the tumor cell biology, the potential of WNT7a and ITGA9 as diagnosis or therapeutic targets for NPC should be considered. LIST OF PUBLICATIONS Articles included in this thesis: 1. Zi-Ming Du, Chun-Fang Hu, Qiong Shao, Ma-Yan Huang, Chang-Wei Kou, Xiao-Feng Zhu, Yi-Xin Zeng, and Jian-Yong Shao*. Upregulation of Caveolin-1 and CD147 Expression in Nasopharyngeal Carcinoma Enhanced Tumor Cell Migration and Correlated with Poor Prognosis of the Patients. Int. J. Cancer: 125, 1832-1841 (2009). PMID: 19582878 2. Zi-Ming Du, Li-Fu Hu, Hai-Yun Wang, Li-Xu Yan, Yi-Xin Zeng, Jian-Yong Shao*, Ingemar Ernberg*. Upregulation of MiR-155 in Nasopharyngeal Carcinoma is partly driven by LMP1 and LMP2A and downregulates a Negative Prognostic Marker JMJD1A. PLoS ONE. 2011 Apr 26;6(4):e19137. PMID: 21541331 3. Zi-Ming Du#, Chang-Wei Kou#, Chun-Fang Hu, Jing Chen, Hai-Yun Wang, Li-Xu Yan, Li-Fu Hu, Ingemar Ernberg, Yi-Xin Zeng and Jian-Yong Shao*. Clinical Significance of Elevated Spleen Tyrosine Kinase Expression in Nasopharyngeal Carcinoma. Head Neck. 2012 Jan 27. PMID: 22287277 4. Imran Nawaz#, Zi-Ming Du#, Ilya Ignatyev#, Tatiana V Pavlova, Vladimir Kashuba, Eugene R Zabarovsky, Ingemar Ernberg, Li-Fu Hu*. WNT7a and ITGA9 Gene are Hypermethylated and Downregulated in Nasopharyngeal Carcinoma. (manuscript) Related publications: 1. Yan Zhang#, Li-Xu Yan#, Qi-Nian Wu#, Zi-Ming Du#, Jing Chen, Ding-Zhun Liao, Ma-Yan Huang, Jing-Hui Hou, Qiu-Liang Wu, Mu-Sheng Zeng, Wen-Lin Huang, Yi-Xin Zeng and Jian-Yong Shao*. miR-125b is Methylated and Functions as A Tumor Suppressor by Regulating the ETS1 proto-oncogene in Human Invasive Breast Cancer. Cancer Res. 2011 May 15;71(10):3552-62. PMID: 21444677 2. Hai-Yun Wang, Bing-Yu Sun, Zhi-Hua Zhu, Ellen T Chang, Ka-Fai To, Jacqueline SG Hwang, Hao Jiang, Michael Koon Ming Kam, Gang Chen, Shie-Lee Cheah, Ming Lee, Zhi-Wei Liu, Jing Chen, Jia-Xing Zhang, Hui-Zhong Zhang, Jie-Hua He, Fa-Long Chen, Xiao-Dong Zhu, Ma-Yan Huang, Ding-Zhun Liao, Jia Fu, Qiong Shao, Man-Bo Cai, Zi-Ming Du, Li-Xu Yan, Chun-Fang Hu, Ho Keung Ng, Joseph TS Wee, Weimin Ye, Ingemar Ernberg, Hans-Olov Adami, Anthony T Chan, Yi-Xin Zeng, and Jian-Yong Shao*. An Eight-signature Classifier for Prediction of Nasopharnyngeal Carcinoma Survival. J Clinl Oncol. 2011 Dec 1;29(34):4516-25. PMID: 22025164 # These authors contributed equally * Corresponding author