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Title: | 2''-5''-寡腺苷酸合成酶3 (OAS3)誘導口腔癌細胞EMT與幹細胞特性之研究 2''-5''-oligoadenylate synthase 3 (OAS3) induced epithelial mesenchymal transition and stemness in oral cancer |
Authors: | 吳庭妤 Ting-Yu Wu |
Advisor: | 鄭世榮 Shih-Jung Cheng |
Co-Advisor: | 郭彥彬 Mark Yen-Ping Kuo |
Keyword: | OAS3,癌幹細胞,檳榔鹼,口腔癌,Stemness,EMT, OAS3,Cancer Stem Cell,Arecoline,Oral Cancer,Stemness,EMT, |
Publication Year : | 2024 |
Degree: | 碩士 |
Abstract: | 根據111年衛生福利部公佈的十大癌症死因死亡率統計,口腔癌高居男性十大癌症死亡率的第四位。抽菸、喝酒與嚼檳榔為台灣口腔癌的主要致病危險因子,其中嚼檳榔比起抽菸更具統計意義。實驗室先前以微陣列分析口腔癌SAS細胞株及具有幹細胞特性的SAS聚球體細胞其基因表現差異,分別以Gene Ontology, KEGG和Protein-Protein Interaction的資料庫分析,篩選出過去並無任何文獻報導與口腔癌有相關性之2'-5'-oligoadenylate synthetase 3 ( OAS3)。我們初步利用OAS3免疫染色發現正常口腔上皮組織(n=12) OAS3陽性染色labeling index(LI)中位數為15%而OSCC患者(n=11)陽性染色中位數為77.5%。顯示OAS3在口腔癌組織有過度表現且具統計意義(p<0.0001)。進一步於細胞實驗中以OAS3質體及空載體轉殖入TW2.6細胞及SAS細胞。發現OAS3過表現的口腔癌細胞Migration和Invasion能力具明顯增強,OAS3過表現的TW2.6細胞及SAS細胞形成聚球體( sphere )的能力明顯提升,並增進口腔癌細胞生長的能力。此外,OAS3過表達會使上皮間質轉換相關蛋白N-Cadherin、Vimentin、Slug、Snail、Twist表現提升,E-Cadherin表現下降,同時增加癌幹細胞(Stemness)相關標誌OCT4、KLF4、SOX2及NANOG的表現。使用OAS3 siRNA knockdown OAS3表現量較多的SAS和FaDu聚球體細胞,發現可降低Stemness相關蛋白表現和形成聚球體的能力。PPI分析中亦發現OAS3與干擾素誘導基因(interferon-stimulated genes, ISGs)的IFIT1及IFIT3有高度相關。OAS3過表現或OAS3 siRNA knockdown的TW2.6細胞中,OAS3的表現與IFIT1和IFIT3的蛋白表現量呈正相關。小鼠頰黏膜異體口腔癌模式結果顯示,OAS3過表現TW2.6細胞腫瘤起始頻率是vector組的10倍以上(p<0.01)。顯示OAS3亦能夠增進口腔癌起始發生。 OAS3過表現TW2.6細胞腫瘤組織OAS3染色表現增加,相對的癌幹細胞及表皮間質標誌表現亦明顯上升,且IFIT1、IFIT3蛋白表現也都有明顯正相關的陽性染色增加。為了解嚼檳榔與OAS3表現的相關性,我們以檳榔鹼Arecoline處理人類口腔上皮SG細胞及TW2.6細胞,發現Arecoline可增加此兩株細胞OAS3的表現量。加入TGF-ß中和抗體、ALK5抑制劑、Smad3抑制劑,可抑制Arecoline誘導OAS3表現。顯示Arecoline經由TGF-ß訊息傳遞路徑誘導口腔上皮細胞OAS3的表現。 從上述結果顯示,Arecoline引發OAS3過表現,可促進頭頸上皮細胞癌化及癌細胞幹性化,其路徑可能經過下游的IFIT1及IFIT3之作用。 The chewing of areca nut (AN, Areca catechu) preparations has been associated with the high incidence of oral cancer observed in Taiwan. Previous studies in our laboratory found 2'-5'-oligoadenylate synthetase 3 (OAS3) is overexpressed in the stem cell-like SAS sphere cells. OAS3 expression in the oral squamous cell carcinoma (OSCC) tissues was higher than those of the normal oral mucosal samples. Human buccal SCC TW2.6 cells and tongue SCC SAS cells with OAS3 overexpression exhibited enhanced migration, invasion, epithelial–mesenchymal transition (EMT), cancer stem cell (CSC) phenotypes. OAS3 overexpression in TW2.6 cells increased tumor initiating frequency at least 10-folds in SCID mice. Furthermore, a positive correlation between OAS3 expression and IFIT1 and IFIT3 overexpression in TW2.6-OAS3 xenograft tissues was observed. Arecoline, a main alkaloid of areca nut, induced the expression of OAS3 protein in oral epithelial SG and TW2.6 cells. Pretreatment with TGF-β neutralizing antibody, SB431542 and smad3 inhibitor SIS3 inhibit the arecoline-induced OAS3 expression, indicating arecoline-induced OAS3 expression is mediated by TGF-β1. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/95074 |
DOI: | 10.6342/NTU202401604 |
Fulltext Rights: | 未授權 |
Appears in Collections: | 口腔生物科學研究所 |
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