Abstract

 
 
 
 Purpose: To determine the regulatory role of sperm-associated antigen 9 (SPAG9) in melanoma cell growth.
 Methods: Immunohistochemical analysis was performed to investigate SPAG9 expression in melanoma tissues. Western blot was used to evaluate SPAG9 expression in melanoma tissues and cells. Melanoma cells were transfected with an siRNA targeting SPAG9 or a SPAG9 overexpression vector. Cell migration and invasion were examined by wound healing and transwell assays. The effects of SPAG9 on the epithelial-mesenchymal transition and mitogen-activated protein kinase (MAPK) pathway in melanoma cells were assessed by western blot.
 Results: SPAG9 expression was enhanced in melanoma tissues and cells. SiRNA-mediated silencing of SPAG9 repressed melanoma cell migration and invasion, and SPAG9 overexpression contributed to melanoma cell metastasis. In melanoma cells transfected with siRNA-SPAG9, E-cadherin expression decreased while vimentin and matrix metalloproteinase (MMP)-2/9 expression increased. However, ectopic expression of SPAG9 reversed this expression of E-cadherin, vimentin, and MMP-2/9. Silencing of SPAG9 decreased the phosphorylation of MAPKs, including p-p38, p-ERK and p-JNK, in melanoma cells. SPAG9 overexpression upregulated phosphorylation of MAPKs.
 Conclusion: SPAG9 promotes the migration and invasion of melanoma cells by activating the MAPK pathway.
 
 
 
Highlights
Melanoma is a highly aggressive cancer that is caused by exposure to ultraviolet radiation [1,2]
To investigate the role of sperm-associated antigen 9 (SPAG9) in melanoma, SPAG9 expression in melanoma tissues was assessed by immunohistochemistry and western blot analysis, and higher SPAG9 expression was found in melanoma tissues than in adjacent normal tissues (Figure 1A, B)
SPAG9 expression was upregulated in the melanoma cell lines A375, G361, SK-MEL-1, and A2058, compared to the human epidermal keratinocyte line HACAT (Figure 1C), suggesting that SPAG9 may participate in melanoma progression
Summary
Melanoma is a highly aggressive cancer that is caused by exposure to ultraviolet radiation [1,2]. Activation of mitogen-activated protein kinases (MAPKs) has been shown to contribute to the epithelial-mesenchymal transformation and extravasation of tumor cells and to promote. Sperm-associated antigen 9 (SPAG9), a cancer/testis antigen, was found to be expressed abnormally in various human malignancies and to play a key role in tumor proliferation and invasion [9]. Silencing of SPAG9 retarded triple-negative breast cancer cell growth and metastasis [13]. Previous studies have shown that SPAG9 binds to c-Jun N-terminal kinases (JNKs) [14] and plays an important regulatory role in several physiologic processes, such as cell survival and tumor development [15]. This study investigated the hypothesis that SPAG9 contributes to melanoma cell metastasis through activation of MAPKs
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