Sprouty2於人類口腔鱗狀細胞癌癌化表現之研究

A Study of Sprouty2 Expression in Human Oral Squamous Cell Carcinogenesis

廖倍顯(Pai-Shien Liao)

指導教授 ： 陳玉昆

共同指導教授 ： 陳丙何(Ping-Ho Chen)

高雄醫學大學/口腔醫學院/牙醫學系博士班/博士(2018年)

摘要

目的 : 本研究探討SPRY2在人類口腔潛在惡性疾病和口腔鱗狀細胞癌中的表現。
方法： 75例口腔鱗狀細胞癌，23例無惡性轉變的口腔潛在惡性疾病，17例有惡性轉變的口腔潛在惡性疾病，8例正常口腔粘膜進行免疫組織化學染色實驗；將具有正常組織對照的3個口腔鱗狀細胞癌組織進行西方墨點法分析。將3種口腔鱗狀細胞癌細胞株，口腔癌前細胞株和正常口腔黏膜初代培養細胞進行西方墨點法分析; 人類口腔鱗狀細胞癌細胞株和正常口腔黏膜初代培養細胞進行即時定量反轉錄聚合酶連鎖反應分析。評估人類口腔鱗狀細胞癌細胞株的增殖，遷移，入侵分析和BRAF V600E點突變檢測。
結果： 口腔鱗狀細胞癌組織中SPRY2蛋白的表現與正常口腔黏膜相比明顯增加，口腔鱗狀細胞癌患者有淋巴結轉移和無淋巴結轉移的SPRY2表現也有差異。與正常口腔黏膜初代培養細胞相比，SPRY2蛋白和mRNA表現顯著增加。在口腔鱗狀細胞癌細胞株中觀察到與正常口腔黏膜初代培養細胞相比增加的磷酸-ERK表現。進行SPRY2 siRNA轉染的口腔鱗狀細胞癌細胞株的增殖率，遷移和入侵的程度與未轉染SPRY2 siRNA的細胞株相比SPRY2蛋白顯著降低。與正常口腔黏膜初代培養細胞相比，在口腔鱗狀細胞癌細胞株中沒有觀察到BRAF V600E點突變。與無惡性轉變的口腔潛在惡性疾病相比，有惡性轉變的口腔潛在惡性疾病患者的SPRY2蛋白水平顯著增加，而與正常口腔黏膜相比，有惡性轉變的口腔潛在惡性質疾病的患者SPRY2蛋白水平顯著增加。與正常口腔黏膜初代培養細胞相比，口腔癌前細胞株SPRY2蛋白水平也有明顯增加。
結論： 實驗結果顯示，SPRY2過度表現與人類口腔鱗狀細胞癌癌化有相關。

關鍵字

SPRY2蛋白 ； 口腔癌癌化 ； 口腔鱗狀細胞癌 ； 口腔潛在惡性疾病 ； 人類

並列摘要

Objective: This study investigated SPRY2 expression in human oral potentially malignant disorders (OPMDs) and oral squamous cell carcinomas (OSCCs).
Methods: 75 OSCCs, 23 OPMDs without malignant transformation (MT), 17 OPMDs with MT, and eight normal oral mucosa (NOM) tissues were used for immunohistochemical staining; three OSCC tissues with normal tissue counterparts were used for western blotting. Three human oral cancer cell lines (OCCLs), an oral precancer cell line (DOK), and a NOM primary culture (NOMPC) were used for western blotting; OCCLs and NOMPC were employed for real-time quantitative reverse transcription-polymerase chain reaction. OCCLs were evaluated in terms of proliferation, migration, invasion and BRAF V600E point mutation assays.
Results: Significantly increased SPRY2 protein expression was observed in OSCCs as compared with NOM, and SPRY2 expression also differed between OSCC patients with and without lymph-node metastasis. SPRY2 protein and mRNA expressions were significantly enhanced as compared with NOMPC. Increased phospho-ERK expression was observed in OCCLs as compared with NOMPC. Significant decreases in the proliferation rate, degrees of migration and invasion were noted in OCCLs with SPRY2 siRNA transfection as compared with those without SPRY2 siRNA transfection. No BRAF V600E point mutation was observed for OCCLs as compared with NOMPC. A significantly increased SPRY2 protein level was noted in OPMDs with MT as compared to those without MT, and was also found in OPMDs with MT in comparison with NOM, as well as in DOK in comparison with NOMPC.
Conclusion: Experimental results indicated that SPRY2 overexpression is associated with human oral squamous-cell carcinogenesis.

並列關鍵字

Sprouty2 ； Oral carcinogenesis ； Oral squamous cell carcinoma ； Oral potentially malignant disorder ； Human

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