Huijuan Liu1, Jie Wang1, Guiyun Ren2, Yanning Zhang1, Fusheng Dong2. 1. Department of Oral Pathology, College and Hospital of Stomatology, Key Laboratory of Stomatology, Hebei Medical University, Hebei Province, China. 2. Department of Oral & Maxillofacial Surgery, College and Hospital of Stomatology, Key Laboratory of Stomatology, Hebei Medical University, Hebei Province, China.
Abstract
OBJECTIVE: To study the relationship between proteoglycan inhibition and the implantation of salivary pleomorphic adenoma (SPA). METHODS: SPA fresh tissue was primitively cultured and identified. The Ad-shRNA-XT-II adenovirus vector was constructed and transfected into SPA cells to silence the XT-II gene. The expression of the XT-II gene and protein was detected using real-time PCR and Western blotting, respectively. The proteoglycan content of the cells was determined 48 h after transfection. The invasion and migration of SPA cells were observed using Matrigel invasion and wound-healing assays. Fibroblasts from the tumour capsule of the same patient were obtained, cultured and seeded onto an acellular dermal matrix (ADM) scaffold. Tumour cells were seeded onto the scaffold with the fibroblasts and then transferred to BALB/C-nu nude mice and allowed to grow in vivo for 3 months. RESULTS: The SPA cells cultures were positive for human calponin, S-100 protein, α-SMA and CK. XT-II gene and protein expression was decreased by 42.72% and 34%, respectively. The proteoglycan content was downregulated by 41.15%. XT-II gene silencing decreased the invasion and migration abilities of SPA cells. The assessment of SPA growth in nude mice indicated an absence of tumour growth in the SPA-XT-II group (in which the XT-II gene was silenced), whereas SPA growth was observed in the other two groups (in which the XT-II gene was not silenced), and the tumour tissue was positive for the human S-100 protein, α-SMA and CK8&18. CONCLUSION: Proteoglycan inhibition induced via XT-II gene silencing inhibited the implantation of SPA.
OBJECTIVE: To study the relationship between proteoglycan inhibition and the implantation of salivary pleomorphic adenoma (SPA). METHODS: SPA fresh tissue was primitively cultured and identified. The Ad-shRNA-XT-II adenovirus vector was constructed and transfected into SPA cells to silence the XT-II gene. The expression of the XT-II gene and protein was detected using real-time PCR and Western blotting, respectively. The proteoglycan content of the cells was determined 48 h after transfection. The invasion and migration of SPA cells were observed using Matrigel invasion and wound-healing assays. Fibroblasts from the tumour capsule of the same patient were obtained, cultured and seeded onto an acellular dermal matrix (ADM) scaffold. Tumour cells were seeded onto the scaffold with the fibroblasts and then transferred to BALB/C-nu nude mice and allowed to grow in vivo for 3 months. RESULTS: The SPA cells cultures were positive for human calponin, S-100 protein, α-SMA and CK. XT-II gene and protein expression was decreased by 42.72% and 34%, respectively. The proteoglycan content was downregulated by 41.15%. XT-II gene silencing decreased the invasion and migration abilities of SPA cells. The assessment of SPA growth in nude mice indicated an absence of tumour growth in the SPA-XT-II group (in which the XT-II gene was silenced), whereas SPA growth was observed in the other two groups (in which the XT-II gene was not silenced), and the tumour tissue was positive for the humanS-100 protein, α-SMA and CK8&18. CONCLUSION: Proteoglycan inhibition induced via XT-II gene silencing inhibited the implantation of SPA.