Jinfeng Shi1, Shulian Tan2, Liangmei Song3, Liangsong Song4, Yusheng Wang1. 1. Department of Otolaryngology Head and Neck surgery, The First Hospital of Jilin University, 130021, Jilin, Changchun, China. 2. Department of Institute of Immunology, The First Hospital of Jilin University, 130021, Jilin, Changchun, China. 3. Department of Operation Room, The First Hospital of Jilin University, 130021, Jilin, Changchun, China. 4. Department of Hand and Foot Surgery, The First Hospital of Jilin University, 130021, Jilin, Changchun, China. Electronic address: xaqxij@163.com.
Abstract
BACKGROUND: Long non-coding RNA (lncRNA) X inactivate-specific transcript (XIST) has been verified as an oncogenic gene in human cancers, including nasopharyngeal carcinoma (NPC). However, the role of XIST in NPC remains to be largely uncovered, as well as its underlying mechanism. METHODS: Expression of XIST, miR-148a-3p and ADAM17 was detected using qPCR and western blot assay. Cell proliferation and apoptosis assay were measured with MTT and flow cytometry, separately. Migration and invasion abilities were examined by transwell assays. Epithelial-mesenchymal transition (EMT) was assessed by western blot analyzing levels of E-cadherin, N-cadherin and vimentin. The potential binding between miR-148a-3p and XIST/ADAM17 was validated by luciferase reporter assay, Ago2-RNA immunoprecipitation and RNA pull-down assay. Xenograft experiments were conducted to measure tumor growth. RESULTS: XIST was upregulated and miR-148a-3p was downregulated in NPC tissues and cell lines. Both XIST knockdown and miR-148a-3p overexpression promoted apoptosis, suppressed cell proliferation, migration, invasion, and EMT of NPC cells in vitro. In addition, miR-148a-3p was validated as a target of XIST, and silencing of miR-148a-3p could reverse XIST knockdown-mediated functions in SUNE-1 and CNE2 cells. Furthermore, miR-148a-3p was identified to target ADAM17, and ectopic expression of ADAM17 could abate miR-148a-3p-induced effects as well. Notably, ADAM17 was downregulated by XIST knockdown through upregulating miR-148a-3p. In vivo, XIST knockdown resulted in a slower tumor growth. CONCLUSION: Knockdown of XIST suppresses the malignant progression of NPC cells through targeting miR-148a-3p/ADAM17 axis both in vitro and in vivo.
BACKGROUND: Long non-coding RNA (lncRNA) X inactivate-specific transcript (XIST) has been verified as an oncogenic gene in humancancers, including nasopharyngeal carcinoma (NPC). However, the role of XIST in NPC remains to be largely uncovered, as well as its underlying mechanism. METHODS: Expression of XIST, miR-148a-3p and ADAM17 was detected using qPCR and western blot assay. Cell proliferation and apoptosis assay were measured with MTT and flow cytometry, separately. Migration and invasion abilities were examined by transwell assays. Epithelial-mesenchymal transition (EMT) was assessed by western blot analyzing levels of E-cadherin, N-cadherin and vimentin. The potential binding between miR-148a-3p and XIST/ADAM17 was validated by luciferase reporter assay, Ago2-RNA immunoprecipitation and RNA pull-down assay. Xenograft experiments were conducted to measure tumor growth. RESULTS:XIST was upregulated and miR-148a-3p was downregulated in NPC tissues and cell lines. Both XIST knockdown and miR-148a-3p overexpression promoted apoptosis, suppressed cell proliferation, migration, invasion, and EMT of NPC cells in vitro. In addition, miR-148a-3p was validated as a target of XIST, and silencing of miR-148a-3p could reverse XIST knockdown-mediated functions in SUNE-1 and CNE2 cells. Furthermore, miR-148a-3p was identified to target ADAM17, and ectopic expression of ADAM17 could abate miR-148a-3p-induced effects as well. Notably, ADAM17 was downregulated by XIST knockdown through upregulating miR-148a-3p. In vivo, XIST knockdown resulted in a slower tumor growth. CONCLUSION: Knockdown of XIST suppresses the malignant progression of NPC cells through targeting miR-148a-3p/ADAM17 axis both in vitro and in vivo.
Authors: Huimin Liu; Dongxu Wang; Shaoning Kan; Ming Hao; Lu Chang; Pengxu Lu; Yangyang Liu; Ye Jin; Weiwei Liu Journal: Front Cell Dev Biol Date: 2022-08-10