論文名稱:

口腔鱗狀細胞癌中FHIT基因的分子變異

 

Molecular Alterations Analysis of the FHIT Gene in Oral Squamous Cell Carcinomas

研究生:

曾若嘉  Ruo-Chia Tseng

 

(以作者名查詢陽明大學館藏系統)

 

(以作者名查詢全國圖書書目資訊網)

指導教授:

張國威  Kuo-Wei Chang

        學位類別:

碩士

        學校名稱:

國立陽明大學

系所名稱:

口腔生物研究所

            學號:

870H02

          學年度:

88

          語文別:

中文

          出版年:

91

關鍵字:

口腔鱗狀細胞癌  oral squamous cell carcinomas

 

FHIT基因  FHIT

 

突變  mutation

 

甲基化  methylation

        論文頁數:

64

摘要:

摘要 FHIT位於染色體3p14.2,在硬塊腫瘤及腫瘤細胞株中常有異常。為證明在口腔鱗狀細胞癌(OSCC)中,FHIT基因及其產物是否發生變異,我們利用聚合酶鏈反應-單股構形多型性,反轉錄-聚合酶鏈反應,亞硫酸基因定序分析及免疫組織化學染色法來偵測在口腔鱗狀細胞癌及非腫瘤配對組織(NCMT)FHIT基因之異常。聚合酶鏈反應-單股構形多型性分析結果顯示未發現FHIT密碼外子有同基因合子之明顯刪除或點突變。利用亞硫酸基因定序分析偵測FHIT基因5’ CpG島叢之甲基化。結果顯示有33% (8/24)口腔鱗狀細胞癌樣本有甲基化之表現。反轉錄-聚合酶鏈反應分析顯示54% (15/28)口腔鱗狀細胞癌樣本表現異常轉錄。在非腫瘤配對組織中未發現有FHIT基因突變,異常轉錄及甲基化情形。利用免疫組織化學染色法偵測Fhit蛋白質表現,分析結果顯示48% (16/33)口腔鱗狀細胞癌樣本其Fhit蛋白質表現量降低。我們的研究結果指出在口腔癌中FHIT基因變異是經常發生以及在口腔鱗狀細胞癌中啟動區之甲基化和轉錄異常可能是導致FHIT失活之重要因素。

   
 

Abstract    Alternations of the FHIT gene at 3p14.2 are frequent in a variety of solid tumors and cancer cell lines. To determine whether the Fhit gene is altered in oral squamous cell carcinomas (OSCC), we examined the gene and its product by RT-PCR, PCR-SSCP, bisulfite genomic sequencing analysis and IHC in OSCCs and non-cancerous match tissues (NCMT). The results indicated that neither homogenous deletion nor point mutation were present in FHIT coding exons by PCR-SSCP method. By bisulfite genomic sequencing, methylation of 5’ CpG islands of the FHIT gene was directly examined. There were 31% (8/26) subjects exhibiting methylation. In addition, 54% (15/28) subjects exhibited aberrant transcripts by RT-PCR method. No mutation, abnormal transcripts or methylation of FHIT was identified in NCMT analyzed. We examined 33 paired OSCCs and NCMT by immunohistochemistry. 48% (16/33) OSCCs exhibited reduction or complete loss of the Fhit expression. No correlation was found between the alterations of FHIT and clinicopathological parameters. Our data indicated that the alteration of FHIT is a frequent event associated with oral cancer formation and suggested methylation at the promoter region and aberrance of transcription were the major underlying causes for FHIT inactivation in OSCC.