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研究生(外文):Hsin-Lan Chen
論文名稱(外文):MIC-A/B expression and regulation in Oral Squamous Cell Carcinoma
指導教授(外文):Kuo-Wei Chang
外文關鍵詞:MICAMICBbiology stressNKG2DMajor histocompatibilitypolymorphism
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近十年來癌症在國人死因中一直名列前矛,癌症的控制仍然有許多問題無法克服,例如抗藥性或癌轉移等問題。因此,發現更有效的癌標誌抗癌藥物或具有癌症預防效果的藥物是敝人感興趣的課題。所以本實驗的目的是想找出口腔鱗狀上皮細胞癌 (Oral squamous cell carcinoma) 之可能診斷的標誌,並探討出具癌症預防效果的藥物對其之抑制。
MICA血中濃度似乎是一種指標。因從腫瘤細胞所脫落的MICA似可調控NKG2D的參與免疫監視反應。MICA及MICB ( Major Histocompatibility Complex class I related Chain A/B ) 是MHC Class I的homologous,而在具有上皮來源的細胞發生壓力(stress) 時產生,而MIC-A/B則為自然殺手細胞(NK cell) 或T細胞的受體NKG2D的ligand。本實驗室先前研究指出MICA之基因多型性與口腔鱗狀上皮細胞癌感受度有關。
利用RT-PCR及費雪氏分析發現,人類正常角化細胞 (NHOK) 與口腔鱗狀上皮癌細胞株及所有的癌細胞株比較,內生性的MICA mRNA的表現並沒有顯著差異。但口腔鱗狀上皮癌細胞株較NHOK於內生性MICB mRNA的表現,顯著的升高 (P = 0.04)。所有的癌細胞株之總表現比NHOK亦有顯著升高 (P = 0.01)。
分析36個口腔鱗狀上皮細胞癌病人的癌組織及癌旁組織發現, MICB的表現是癌組織大於癌旁組織共佔56%,MICA佔50%,利用paired-t test分析的結果,MICB的癌組織大於癌旁組織於統計上是有意義的 (P = 0.009)。在與臨床與病理發現交叉分析顯示,MICA的mRNA表現量與患者嚼食檳榔,有顯著相關 (P = 0.001)。與腫瘤細胞分化有顯著相關 (P = 0.02)。分析患者年齡、腫瘤發生部位、淋巴結轉移以及癌症分期等並無明顯相關。這種臨床意義首次在口腔鱗狀上皮細胞癌內的發現。
以可有效抑制口腔癌細胞生長的綠茶多酚物質主成份EGCG,處理口腔癌細胞株SAS,24、48、72小時,MICA及MICB mRNA表現, 在24及72小時有明顯抑制的效果。薑黃素來處理SAS亦發現5μM薑黃素於48小時,可抑制MICA及MICB mRNA表現。
以檳榔子水萃取液5μg/ml處理SAS 24小時,則發現可使得MICA及MICB mRNA表現皆輕度上升。
本研究初步結論MICA及MICB mRNA表現之升高可為口腔鱗狀上皮細胞癌之可能標誌,且MICA及MICB mRNA亦可為抗癌物質調降,具探討其應用之價值。
Malignancy has been the top cause of mortality in our country. Problems of diagnosis and resolution remain to be overcomed. Therefore, it is very important to study more efficient anti-cancer drugs. The aims of the present study was to identify molecular alteration and efficant anti-cancer modalities.The aim was to find out a useful molecular marker and its inhibitor.
The concentration of MICA in serum seems be a good candidate for marker, because the soluble MICA derived from tumor can regulate the immune surveillance involvement of NKG2D. MICA and MICB (Major Histocompatibility Complex class I related Chain A/B) were homologous of MHC Class I. They are induced when epithelial cells evoked by stress. MICA and MICB when binding to NK cell or T cell receptor, is NKG2D’s ligand。Previous studies from our laboratory have shown that the polymorphic of MICA gene might confer the risk of oral squamous cell carcinoma (OSCC)
RT-PCR analysis and Fisher’s exact test indicated that endogenos MICA mRNA expression is not significantly different when comparing NHOK to OSCC cell lines、breast and lung cancer cell lines.
Endogenos MICB mRNA expression in OSCC cell lines was significantly higher than that in NHOK (P = 0.04)。OSCC cell lines、breast and lung cancer cell lines also have MICB mRNA expression higher than NHOK (P = 0.01)。Analyses of 36 paired OSCCs and NCMTs showed that the MICB mRNA expression in OSCC is higher than NCMT in 56% cases。Paired t-test also showed that MICB mRNA expression in OSCC (0.39 ± 0.08) was significant higher than that in NCMT (P = 0.009). Interestingly, MICA mRAN expression is associated with chewing area nut (P = 0.001) and tumor differentiation (P = 0.02). Age, site, lymph node metastasis and clinical stage were not significantly correlated MICA and MICB mRNA expression. This is the first study of MICA and MICB mRNA expression in OSCC.
EGCG can effectively inhibit OSCC cell growth and to treat SAS cells, the MICA and MICB mRNA expression is significant down-regulated at 24 and 72 h. Curcumin inhibits MICA and MICB mRNA expression. Using 5μg/ml ANE to treat SAS cells for 24 h, MICA and MICB mRNA expression was slightly increased.
Our findings suggest increase of MICB mRNA expression OSCC could be used as a marker.The regulation of EGCG, Curcumin and ANE on MICA and MICB mRNA expression could suggest a therapeutic application.
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