JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE) ›› 2021, Vol. 41 ›› Issue (6): 717-723.doi: 10.3969/j.issn.1674-8115.2021.06.003

• Basic research • Previous Articles     Next Articles

Impact of Ras association domain family 5 on cell migration and invasion of head and neck squamous cell carcinoma

Yu-sheng LU(), Wen-yi YANG, Hai-long MA(), Jing-zhou HU()   

  1. Department of Oral Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology, Shanghai 200011, China
  • Online:2021-06-28 Published:2021-06-29
  • Contact: Hai-long MA,Jing-zhou HU;;
  • Supported by:
    National Natural Science Foundation of China(81872185);Program of Innovative Research Team of High-level Local Universities in Shanghai(SSMU-ZLCX20180502)

Abstract: Objective

·To investigate the effect of Ras association domain family 5 (RASSF5) on migration and invasion of head and neck squamous cell carcinoma (HNSCC).


·The expression of RASSF5 gene in 563 cases of HNSCC from the Cancer Genome Atlas (TCGA) database and the effect of RASSF5 expression on prognosis and survival of patients were analyzed by Gene Expression Profiling Interactive Analysis (GEPIA) platform. The lentivirus overexpressing RASSF5 (LV RASSF5) and control lentivirus (LV vector) were constructed by CMV-MCS-PGK-Puro vector. The wound-healing assay and the Transwell assay were performed to test the invasion and metastasis capacity of overexpressed-RASSF5 HNSCC cells. In addition, the expressions of epithelial cadherin (E-cadherin), Snail, glycogen synthase kinase 3β (GSK-3β) and phosphorylated GSK-3β (p-GSK-3β) were determined by Western blotting in RASSF5-overexpressed HNSCC cells. The lung metastasis model of HNSCC in 10 BALB/c nude mice were established. Cal27, which were transfected with LV RASSF5 or LV vector, were respectively injected into 5 BALB/c nude mice via tail vein, and the quantity of metastasis nodules was counted to evaluate the effect of RASSF5 overexpression on tumor metastasis ability.


·In HNSCC, the gene expression level of RASSF5 was lower than that in normal tissues (P=0.001). Patients with lower gene expression of RASSF5 had worse overall survival (P=0.005) and disease-free survival (P=0.004). The relative healing areas of Cal27 and HN30 transfected with LV RASSF5 were higher than those of the control group (transfected with LV vector) at the experimental endpoint of the wound-healing assay (P=0.015, P=0.003). The number of cells that had traversed the cell-permeable membrane was higher in Cal27 and HN30 cells transfected with LV RASSF5 than those in the control group in the Transwell migration assay (P=0.005, P=0.001). And in the Transwell invasion assay, the number of invaded cells that had traversed the matrigel-coated membrane was also higher in Cal27 and HN30 cells transfected with LV RASSF5 than those in the control group (P=0.001, P=0.001). Western blotting showed an increased level of E-cadherin, and decreased level of Snail and p-GSK-3β/GSK-3β ratio in RASSF5 overexpression Cal27 and HN30 cells. And in the metastasis assay of BALB/c nude mice, the quantity of metastases nodes was lower in the RASSF5-overexpressed group than that in the control group (P=0.049).


·RASSF5 may inhibit epithelial-mesenchymal transition through GSK-3β/Snail pathway, and then inhibits the invasion and metastasis ability of HNSCC cells.

Key words: head and neck cancer, Ras association domain family 5 (RASSF5), cell migration, invasion, epithelial-mesenchymal transition

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