臺灣博碩士論文加值系統

標靶治療是目前臨床上治療癌症重要的方法，目前已利用表皮生長因子接受器(epidermal growth factor receptor；EGFR)特異性抗體在大腸直腸癌的治療，但有高達 50％病人對這種療法反應不佳。會導致此種標靶治療失敗的原因，可能是因EGFR dependent訊息路徑之相關基因發生突變所導致。本研究於篩選的182位大腸直腸癌檢體，利用高解析度溶解曲線分析法(high resolution melting analysis)分析方法，針對EGFR dependent訊息路徑中可能會發生突變的相關基因位點 (BRAF和PIK3CA基因)，檢測其熱點突變 (hotspot mutation)，再利用直接核酸定序來確認 HRMA的結果。在這182位大腸直腸癌中檢測到28.6% 的KRAS (codon 12, 13)， 1.1% 的BRAF (V600E) ， 2.2% 的PIK3CA (exon 9, 20 ) 及4.9% KRAS與PIK3CA同時發現突變。結果顯示高解析度溶解曲線分析法為一種有效的方法，可快速、高敏感性、有效地檢測 BRAF及PIK3CA基因的熱點突變。在大腸直腸癌KRAS wild-type的病患，利用單株抗體進行治療失敗的原因，可能是BRAF或PIK3CA突變或是intratumoral heterogeneity造成。

Epidermal growth factor receptor (EGFR) monoclonal antibody therapy is established in patients with wild-type KRAS colorectal cancer ; however, up to 50% of these patients do not respond to this therapy. To identify the possible causes of this treatment failure, we searched for mutations of several oncogenes involving in the EGFR-dependent signaling pathways. In this study, high resolution melting analysis (HRMA) was used to screen hot-spot mutations in the BRAF and PIK3CA genes in 182 colorectal cancer specimens. Direct sequencing was used to confirm HRMA results. Activating mutations were detected in 28.6% of KRAS (codon 12, 13), 1.1% of BRAF (V600E), 4.9% of PIK3CA (exon 9, 20) and 4.9% (KRAS and PIK3CA) of the 182 colorectal cancer specimens. HRMA provides a fast, high sensitivity, and valid approach to efficiently detect these oncogene mutations. Failure of EGFR antibody therapy in patients with wild-type KRAS colorectal cancer may result from activating BRAF or PIK3CA mutations.

目錄
1.摘要---------------------------------------------3
2.Abstract-----------------------------------------4
3.背景---------------------------------------------5
4.研究材料與方法----------------------------------12
5.研究結果----------------------------------------33
6.討論--------------------------------------------35
7.附表與附圖--------------------------------------42
8.參考文獻----------------------------------------54

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