OBJECTIVE: To demonstrate the subcellular localization of nasopharyngeal carcinoma (NPC)-associated gene 6 (NGX6) and its basic structure and function. METHODS: The deletion mutants of NGX6 were constructed by one-step PCR and transfected into NPC cell line 5-8F. The subcellular location of NGX6 or its mutants was detected by immunofluorescence staining of cytoplasm (CYTO) and nuclear protein, and immunoelectron-microscopic analysis. The role of NGX6 and its mutants in the proliferation, adhesion and migration of NPC 5-8F cells was detected using the following assays: growth curve, colony formation in soft agar, cell adhesion, in vitro Matrigel invasion, and in vitro scratch wound healing. RESULTS AND CONCLUSIONS: NGX6 and its mutants were distributed on the plasma membrane, nuclear membrane, endoplasmic reticulum membrane, and other membrane structures in the cytosol. The deleted domains did not affect its distribution in 5-8F cells. NGX6 could increase the adhesion but inhibited the proliferation, growth and migration of 5-8F cells. The epidermal growth factor-like domain and CYTO region were found to be important for NGX6 to modulate cell adhesion, and CYTO was found to be essential for NGX6 involved in the regulation of growth, proliferation, and migration of 5-8F cells.
OBJECTIVE: To demonstrate the subcellular localization of nasopharyngeal carcinoma (NPC)-associated gene 6 (NGX6) and its basic structure and function. METHODS: The deletion mutants of NGX6 were constructed by one-step PCR and transfected into NPC cell line 5-8F. The subcellular location of NGX6 or its mutants was detected by immunofluorescence staining of cytoplasm (CYTO) and nuclear protein, and immunoelectron-microscopic analysis. The role of NGX6 and its mutants in the proliferation, adhesion and migration of NPC 5-8F cells was detected using the following assays: growth curve, colony formation in soft agar, cell adhesion, in vitro Matrigel invasion, and in vitro scratch wound healing. RESULTS AND CONCLUSIONS:NGX6 and its mutants were distributed on the plasma membrane, nuclear membrane, endoplasmic reticulum membrane, and other membrane structures in the cytosol. The deleted domains did not affect its distribution in 5-8F cells. NGX6 could increase the adhesion but inhibited the proliferation, growth and migration of 5-8F cells. The epidermal growth factor-like domain and CYTO region were found to be important for NGX6 to modulate cell adhesion, and CYTO was found to be essential for NGX6 involved in the regulation of growth, proliferation, and migration of 5-8F cells.