Archives of Otolaryngology and Rhinology Citation: Yang JH, Lu CS, Zhang S (2015) Alteration of the Methylation Status of Urokinase Plasminogen Activator (uPA) is involved in Proliferation and Invasion of Nasopharyngeal Cancer Cells. Arch Otolaryngol Rhinol 1(1): 023-027. 023 Abstract The extracellular matrix degradation is the most important step in the process of tumor cell metastasis. Urokinase-type plasminogen activator (uPA) can catalyzes the conversion of the inactive zymogen plasminogen to the active broad-spectrum plasmin, which degrades a number of matrix proteins and also activates other proteases, including some matrix metalloproteinases. In this study, we evaluated that effect of methylation status of urokinase plasminogen activator (uPA) on invasion and metastasis of nasopharyngeal cancer cells. Methylation status of urokinase plasminogen activator (uPA) was detected by Methylation-speciic PCR in the nasopharyngeal carcinoma cells. Reverse transcription-PCR was used to detect the expression of uPA mRNA in the nasopharyngeal carcinoma cell. Invasive and proliferation capacity of CNE2 cell was detected by using Boyden chamber Matrigel invasion assay and Cell proliferation assay after MBD2 knockdown. Our data demonstrated that uPA gene promoter hypomethylation is related to strong uPA mRNA expression in nasopharyngeal cancer cell, uPA mRNA expression was signiicantly reduced after MBD2 knockdown in the CNE2 cells, the proliferation and invasion capacity of CNE2 cells was signiicantly inhibited after MBD2 knockdown. Our data suggest that reversal of uPA gene hypomethylation will become a novel therapeutic approach for blocking nasopharyngeal cancer progression and metastatic. role in nasopharyngeal carcinoma progression and metastasis [6]. Recent studies have found that the hypomethylation of oncogene plays a causal role in tumor metastasis. Pakneshan found that hypomethylation is responsible for uPA activation and plays a role in metastasis, inhibition of demethylation result in methylation and inactivation of uPA and reversal of metastasis in breast cancer [7]. In the present study, we demonstrate that methylation state and expression of uPA gene, and its hypomethylated state could be reversed pharmacologically by agents that modulate methylation like an antisense inhibitor of Methyl-binding protein 2 (MDB2) in nasopharyngeal cancer cell line CNE2. his study conirms that uPA is activated by hypomethylation in nasopharyngeal cancer cells, and uPA plays a important role in proliferation and invasion of nasopharyngeal cancer cell. It suggests that pharmacological reversal of uPA gene hypomethylation can be used as a novel therapeutic approach for blocking nasopharyngeal cancer progression. Materials and Methods Cell culture and treatment Human nasopharyngeal cancer cell line CNE2 was obtained from the China Center for Type Culture Collection (CCTCC). CNE2 cell lines were maintained in RPMI 1640 with 10% Fetal bovine serum (FBS), 2 mM L-glutamine adjusted to contain 1.5 g/l sodium bicarbonate, 4.5 g/l glucose, 10 mM HEPES, and 1.0 mM sodium pyruvate. Cells were incubated at 37°C in 5% CO2. Introduction A major feature of cancer cells is their ability to migrate and to invade and develop in surrounding or distant tissues. Tumor cells metastasis to distant organs involves four major steps [1]: adhesion of tumor cells to the extracellular matrix (ECM), ability of tumor cells to degrade the ECM and intravasate into surrounding blood vessels, survival against the natural host defenses and settling at the preferred organ site, and extravasation into the organ and formation of new tumors. he extracellular matrix degradation is the most important step in the process of tumor cell metastasis. Urokinase- type plasminogen activator (uPA) is a member of the serine protease family. It catalyzes the conversion of the inactive zymogen plasminogen to the active broad-spectrum plasmin, which degrades a number of matrix proteins and also activates other proteases, including some matrix metalloproteinases [2]. It has been implicated that uPA plays an important role in the process of invasion, metastasis and angiogenesis of several malignancies [3-5]. Nasopharyngeal cancer (NPC) is a common tumor in head and neck. It has an unusually high incidence in southern China. Cervical lymph node metastasis is a predominant clinical characteristic in the earlier period of nasopharyngeal cancer. It is not very clear that the mechanism of nasopharyngeal cancer metastasis. We have previous study showed that methylation status of Death-associated protein kinase (DAPK), a tumor suppress gene, plays an important Research Article Alteration of the Methylation Status of Urokinase Plasminogen Activator (uPA) is involved in Proliferation and Invasion of Nasopharyngeal Cancer Cells Ju-Hong Yang 1 , Chun-Sheng Lu 1 and Song Zhang 2 * 1 Department of blood transfusion, Guang Ming New District People’s Hospital of Shenzhen, Shenzhen, Guang Dong, TX 518106, PR China 2Department of Otolaryngology, Guang Ming New District People’s Hospital of Shenzhen, Shenzhen, Guang Dong, TX 518106, PR China Dates: Received: 22 May, 2015; Accepted: 06 July, 2015; Published: 09 July, 2015 *Corresponding author: Song Zhang, Department of Otolaryngology, Guang Ming New District People’s Hospital of Shenzhen, Shenzhen, Song Bai Road 339#, Guang Dong, TX 518106, PR China; E-mail: www.peertechz.com Keywords: Nasopharyngeal cancer; Urokinase plasminogen activator (uPA) ; Methyl binding domain protein 2 (MDB2) ; siRNA; CNE2 cell line; Methylation