Investigation of bone invasion and underlying mechanisms of oral cancer using a cell line-derived xenograft model

Qiusheng Shan; Kiyofumi Takabatake; Haruka Omori; Hotaka Kawai; May Wathone Oo; Shintaro Sukegawa; Masae Fujii; Yasunori Inada; Sho Sano; Keisuke Nakano; Hitoshi Nagatsuka

doi:10.3892/ol.2022.13502

Investigation of bone invasion and underlying mechanisms of oral cancer using a cell line-derived xenograft model

Oncol Lett. 2022 Sep 13;24(5):382. doi: 10.3892/ol.2022.13502. eCollection 2022 Nov.

Authors

Qiusheng Shan^{1

2}, Kiyofumi Takabatake¹, Haruka Omori¹, Hotaka Kawai¹, May Wathone Oo¹, Shintaro Sukegawa^{1

3}, Masae Fujii¹, Yasunori Inada¹, Sho Sano^{1

2}, Keisuke Nakano¹, Hitoshi Nagatsuka¹

Affiliations

¹ Department of Oral Pathology and Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Okayama 700-8525, Japan.
² Department of Oral and Maxillofacial Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Okayama 700-8525, Japan.
³ Department of Oral and Maxillofacial Surgery, Kagawa Prefectural Central Hospital, Takamatsu, Kagawa 760-8557, Japan.

Abstract

The cancer stroma regulates bone invasion in oral squamous cell carcinoma (OSCC). However, data on normal stroma are limited. In the present study, the effects of gingival and periodontal ligament tissue-derived stromal cells (G-SCs and P-SCs, respectively) and human dermal fibroblasts (HDFs) on bone resorption and osteoclast activation were assessed using hematoxylin and eosin and tartrate-resistant acid phosphatase staining in a cell line-derived xenograft model. The results demonstrated that G-SCs promoted bone invasion and osteoclast activation and inhibited osteoclast proliferation following crosstalk with the human OSCC HSC-3 cell line, whereas P-SCs inhibited bone resorption and promoted osteoclast proliferation in vitro but had a minimal effect on osteoclast activation both in vitro and in vivo following crosstalk with HSC-3 cells. Furthermore, the effects of G-SCs, P-SCs and HDFs on protein expression levels of matrix metalloproteinase (MMP)-9, membrane type 1 MMP (MT1-MMP), Snail, parathyroid hormone-related peptide (PTHrP) and receptor activator of NF-κB ligand (RANKL) in HSC-3 cells in OSCC bone invasion regions were assessed using immunohistochemistry. The results demonstrated that G-SCs had a more prominent effect on the expression of MMP-9, MT1-MMP, Snail, PTHrP, and RANKL, whereas P-SCs only promoted RANKL and PTHrP expression and exerted a minimal effect on MMP-9, MT1-MMP and Snail expression. The potential genes underlying the differential effects of G-SCs and P-SCs on bone invasion in OSCC were evaluated using a microarray, which indicated that cyclin-dependent kinase 1, insulin, aurora kinase A, cyclin B1 and DNA topoisomerase II alpha underlaid these differential effects. Therefore, these results demonstrated that G-SCs promoted bone invasion in OSCC by activating osteoclasts on the bone surface, whereas P-SCs exerted an inhibitory effect. These findings could indicate a potential regulatory mechanism for bone invasion in OSCC.

Keywords: bone invasion; gingival ligament tissue-derived stromal cell; microarray; oral squamous cell carcinoma; periodontal ligament tissue-derived stromal cell; xenograft model.

Grants and funding

The present study was funded by the Japan Society for Promotion of Science KAKENHI Grants-in-Aid for Scientific Research (grant nos. JP20K10094, JP21K10043, JP21K17089, JP19K19159, JP20H03888 and JP22K10170).