Journal of Shanghai Jiao Tong University (Medical Science) >

2021 , Vol. 41 >Issue 6: 717 - 723

DOI: https://doi.org/10.3969/j.issn.1674-8115.2021.06.003

Basic research

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
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  • 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 published: 2021-06-29

Supported by

National Natural Science Foundation of China(81872185);Program of Innovative Research Team of High-level Local Universities in Shanghai(SSMU-ZLCX20180502)

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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).

Methods

·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.

Results

·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).

Conclusion

·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

Cite this article

Yu-sheng LU , Wen-yi YANG , Hai-long MA , Jing-zhou HU . Impact of Ras association domain family 5 on cell migration and invasion of head and neck squamous cell carcinoma[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2021 , 41(6) : 717 -723 . DOI: 10.3969/j.issn.1674-8115.2021.06.003

References

1 Chow LQM. Head and neck cancer[J]. N Engl J Med, 2020, 382(1): 60-72.
2 Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020[J]. CA A Cancer J Clin, 2020, 70(1): 7-30.
3 Cohen EEW, Soulières D, Le Tourneau C, et al. Pembrolizumab versus methotrexate, docetaxel, or cetuximab for recurrent or metastatic head-and-neck squamous cell carcinoma (KEYNOTE-040): a randomised, open-label, phase 3 study[J]. The Lancet, 2019, 393(10167): 156-167.
4 Barnoud T, Schmidt ML, Donninger H, et al. The role of the NORE1A tumor suppressor in oncogene-induced senescence[J]. Cancer Lett, 2017, 400: 30-36.
5 Donninger H, Schmidt ML, Mezzanotte J, et al. Ras signaling through RASSF proteins[J]. Semin Cell Dev Biol, 2016, 58: 86-95.
6 Zhou XH, Yang CQ, Zhang CL, et al. RASSF5 inhibits growth and invasion and induces apoptosis in osteosarcoma cells through activation of MST1/LATS1 signaling[J]. Oncol Rep, 2014, 32(4): 1505-1512.
7 Liu LL, Zhang MF, Pan YH, et al. NORE1A sensitises cancer cells to sorafenib-induced apoptosis and indicates hepatocellular carcinoma prognosis[J]. Tumor Biol, 2014, 35(3): 1763-1774.
8 Steinmann K, Sandner A, Schagdarsurengin U, et al. Frequent promoter hypermethylation of tumor-related genes in head and neck squamous cell carcinoma[J]. Oncol Rep, 2009, 22(6): 1519-1526.
9 Han L, Dong Z, Wang C, et al. Decreased expression and aberrant methylation of RASSF5A correlates with malignant progression of gastric cardia adenocarcinoma[J]. Mol Carcinog, 2015, 54(12): 1722-1733.
10 Liu G, Liu B, Zheng S, et al. Aberrant RASSF5 gene transcribed region hypermethylation in pediatric hepatoblastomas[J]. Int J Clin Exp Pathol, 2018, 11(7): 3612-3617.
11 Guo W, Wang C, Guo YL, et al. RASSF5A, a candidate tumor suppressor, is epigenetically inactivated in esophageal squamous cell carcinoma[J]. Clin Exp Metastasis, 2015, 32(1): 83-98.
12 Zhou BP, Deng J, Xia WY, et al. Dual regulation of Snail by GSK-3β-mediated phosphorylation in control of epithelial-mesenchymal transition[J]. Nat Cell Biol, 2004, 6(10): 931-940.
13 Sun J, Meng D, Yu T, et al. N-terminal truncated carboxypeptidase E represses E-cadherin expression in lung cancer by stabilizing the Snail-HDAC complex[J]. Am J Cancer Res, 2020, 10(3): 925-938.
14 An P, Chen F, Li Z, et al. HDAC8 promotes the dissemination of breast cancer cells via AKT/GSK-3β/Snail signals[J]. Oncogene, 2020, 39(26): 4956-4969.
15 Domoto T, Pyko IV, Furuta T, et al. Glycogen synthase kinase-3β is a pivotal mediator of cancer invasion and resistance to therapy[J]. Cancer Sci, 2016, 107(10): 1363-1372.
16 Jin F, Wu Z, Hu X, et al. The PI3K/Akt/GSK-3β/ROS/eIF2B pathway promotes breast cancer growth and metastasis via suppression of NK cell cytotoxicity and tumor cell susceptibility[J]. Cancer Biol Med, 2019, 16(1): 38-54.
17 Liao TJ, Tsai CJ, Jang H, et al. RASSF5: an MST activator and tumor suppressor in vivo but opposite in vitro[J]. Curr Opin Struct Biol, 2016, 41: 217-224.
18 Donninger H, Calvisi DF, Barnoud T, et al. NORE1A is a Ras senescence effector that controls the apoptotic/senescent balance of p53 via HIPK2[J]. J Cell Biol, 2015, 208(6): 777-789.
19 Li BT, Yu C, Xu Y, et al. TET1 inhibits cell proliferation by inducing RASSF5 expression[J]. Oncotarget, 2017, 8(49): 86395-86409.
20 Sun HY, Yang D, Mi J, et al. Histone demethylase Jmjd3 modulates osteoblast apoptosis induced by tumor necrosis factor-α through directly targeting RASSF5[J]. Connect Tissue Res, 2020, 61(6): 517-525.
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