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題名 | Application of Methylation-Specific Polymerase Chain Reaction to Investigate Methylation Status of the P16 Promoter CpG Island in Nasopharyngeal Carcinoma=利用甲基化特異巢式反應來偵測鼻咽癌P16基因起始段內CpG小島甲基化情形 |
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作者 | 黃立志; 許永祥; 高瑞和; Huang, Li-chih; Hsu, Yung Hsiang; Kao, Ruey Ho; |
期刊 | 中華民國癌症醫學會雜誌 |
出版日期 | 20010900 |
卷期 | 17:3 2001.09[民90.09] |
頁次 | 頁1-6 |
分類號 | 416.879 |
語文 | eng |
關鍵詞 | 甲基化特異巢式反應; 鼻咽癌; P16基因; CpG小島甲基化; Nasopharyngeal carcinoma; p16; Methylation; Polymerase chain reaction; |
中文摘要 | 目的:本研究利用一種新的技術,甲基化特異巢式反應來測試鼻咽癌組織內p16基因起始段內CpG小島因基化情形,來探討此一機轉在降低p16表現率之角色。 材料與方向:我們採集十五位鼻咽癌病人的石臘組織切片以及三種乳癌細胞株,粹取其DNA,經過重度硫酸鈉處理後,利用特殊之引子進行甲基化特異巢式反應,所得產物跑在膠片上,在紫外光下觀察。 結果:在十五個石臘組織切片中有七個成功的粹取出DNA,其中有五個在甲基化特異巢式反應中跑出產物:兩個(40%)在甲基化巢式反應,三個(60%)在無甲基化巢式反應。剩下兩個在兩種反應中都無產物。 結論:甲基化特異巢式反應確實可應用在鼻咽癌石臘組織切片上。但未來的方向應朝向如何增加鼻咽癌的DNA產量上,使實驗能更完整進行。 |
英文摘要 | Objective: The current study utilized methylation-specific polymerase chain reaction to investigate methylation status of CpG islands on p16 promoter for nasopharyngeal carcinoma (NPC) in order to verify the role of the mechanism in silencing the p16 gene. Material and Methods: Fifteen cases of NPC paraffin-embedded tissue sections along with 3 breast cancer cell lines were subjected to DNA extraction, sodium bisulfite modification and PCR amplification with primers specific for methylated and unmethylated CpG islands on the p16 promoter. The PCR products were run on an agarose gel in the presence of ethidium bromide and visualised under UV illumination. Results: In 15 cases of NPC paraffin-embedded tissue sections, 7 cases were able to be extreated DNAs. Successful methylation-specific-PCR was achieved from 5 of them, among which 2 (40%) showed methylation pattern, 3 (60%) showed unmethylation patterns on CpG islands of the p16 promoter. Conclusion: Methylation-specific PCR is applicable in studying methylation status of CpG islands on p16 promoter for NPC paraffin-embedded tissue section. However effort must be further focused on increasing the throughput of DNA extraction from the preserved tissue material in a study of NPC. |
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