應用酶切富集PCR 法檢測非小細胞肺癌患者胸腔積液表皮生長因子受體基因突變_《中國呼吸與危重監護雜志》

作者：

何臣 , 劉明 ,  徐軍

廣州醫學院第一附屬醫院廣州呼吸疾病研究所呼吸病國家重點實驗室（廣東廣州 510120）;

關鍵詞：

酶切富集PCR 法胸腔積液表皮生長因子受體點突變非小細胞肺癌表皮生長因子受體酪氨酸激酶抑制劑吉非替尼

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目的探討酶切富集PCR 法檢測非小細胞肺癌( NSCLC) 患者胸腔積液表皮生長因子受體基因( EGFR) 外顯子19 缺失和外顯子21 L858R 突變的臨床意義。方法采用EGFR 基因突變酶切富集及非酶切富集PCR 法對30 例NSCLC 患者胸腔積液游離核酸EGFR 基因外顯子19 缺失和外顯子21 L858R 突變進行分析。結果 30 例NSCLC 患者中, 酶切富集PCR 法分別檢出EGFR 基因外顯子19 缺失10 例( 33. 3% ) 和外顯子21 L858R 突變5 例( 1. 7% ) , 非酶切富集PCR 法則僅能分別檢出6 例( 20. 0%) 和1 例( 3. 3% ) ; 兩種方法檢出率差異有統計學意義( P = 0. 032) 。在接受吉非替尼治療的4 例患者中, 2 例檢出EGFR 基因突變患者治療后腫瘤均部分緩解。結論酶切富集PCR 法可以高效、經濟、準確地檢測NSCLC 患者胸腔積液游離核酸EGFR 基因突變, 可能成為臨床NSCLC 患者選擇EGFR 酪氨酸激酶抑制劑治療的預測方法。

引用本文： 何臣,劉明,徐軍. 應用酶切富集PCR 法檢測非小細胞肺癌患者胸腔積液表皮生長因子受體基因突變. 中國呼吸與危重監護雜志, 2009, 09(5): 451-455. doi: 復制

1.	Sharma SV, Bell DW, Settleman J, et al. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer, 2007, 7: 169-181.
2.	Benlloch S, Galbis-Caravajal JM,Martin C, et al. Potential diagnostic value of methylation profile in pleural fluid and serum from cancer patients with pleural effusion. Cancer, 2006, 107: 1859-1865.
3.	Nakamoto M, Teramoto H, Matsumoto S, et al. K-ras and rho A mutations in malignant pleural effusion. Int J Oncol, 2001, 19 : 971 - 976.
4.	Lee JH, Hong YS, Ryu JS, et al. P53 and FHIT mutations and microsatellite alterations in malignancy-associated pleural effusion. Lung Cancer, 2004, 44: 33-42.
5.	Zhang X, Zhao Y, Wang M, et al. Detection and comparison of epidermal growth factor receptor mutations in cells and fluid of malignant pleural effusion in non-small cell lung cancer. Lung Cancer, 2008, 60: 175-182.
6.	Kahn SM, Jiang W, Culbertson TA, et al. Rapid and sensitive nonradioactive detection of mutant K-ras genes via ‘enriched’PCR amplification. Oncogene, 1991 , 6: 1079 -1083 .
7.	Chen J, Viola MV. A method to detect ras point mutations in small subpopulations of cells. Anal Biochem, 1991, 195: 51-56.
8.	Toyooka S, Tsukuda K, OuchidaM, et al. Detection of codon 61 point mutations of the K-ras gene in lung and colorectal cancers by enriched PCR. Oncol Rep, 2003, 10: 1455-1459.
9.	Asano H, Toyooka S, Tokumo M, et al. Detection of EGFR gene mutation in lung cancer by mutant-enriched polymerase chain reaction assay. Clin Cancer Res, 2006, 12: 43-48.
10.	Soh J, Toyooka S, Aoe K, et al. Usefulness of EGFR mutation screening in pleural fluid to predict the clinical outcome of gefitinib treated patients with lung cancer. Int J Cancer, 2006 , 119: 2353 - 2358.
11.	Kimura H, Fujiwara Y, Sone T, et al. EGFR mutation status in tumour-derived DNA from pleural effusion fluid is a practical basis for predicting the response to gefitinib. Br J Cancer, 2006, 95: 1390- 1395.
12.	Kimura H, Fujiwara Y, Sone T, et al. High sensitivity detection of epidermal growth factor receptor mutations in the pleural effusion of non-small cell lung cancer patients. Cancer Sci, 2006, 97: 642-648.
13.	Wu SG, Gow CH, Yu CJ, et al. Frequent epidermal growth factor receptor gene mutations in malignant pleural effusion of lung adenocarcinoma. Eur Respir J, 2008, 32: 924-930.
14.	Kimura H, Kasahara K, Kawaishi M, et al. Detection of epidermal growth factor receptor mutations in serum as a predictor of the response to gefitinib in patients with non-small-cell lung cancer. Clin Cancer Res, 2006, 12: 3915-3921.
15.	Kimura H, Suminoe M, Kasahara K, et al. Evaluation of epidermal growth factor receptor mutation status in serum DNA as a predictor of response to gefitinib( IRESSA) . Br J Cancer, 2007, 97: 778-784.
16.	董強剛, 黃進肅, 韓寶惠, 等. 晚期肺癌患者血清游離DNA 中 EGFR 外顯子19 的基因突變研究. 腫瘤, 2006, 26: 59-63.

1. Sharma SV, Bell DW, Settleman J, et al. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer, 2007, 7: 169-181.
2. Benlloch S, Galbis-Caravajal JM,Martin C, et al. Potential diagnostic value of methylation profile in pleural fluid and serum from cancer patients with pleural effusion. Cancer, 2006, 107: 1859-1865.
3. Nakamoto M, Teramoto H, Matsumoto S, et al. K-ras and rho A mutations in malignant pleural effusion. Int J Oncol, 2001, 19 : 971 - 976.
4. Lee JH, Hong YS, Ryu JS, et al. P53 and FHIT mutations and microsatellite alterations in malignancy-associated pleural effusion. Lung Cancer, 2004, 44: 33-42.
5. Zhang X, Zhao Y, Wang M, et al. Detection and comparison of epidermal growth factor receptor mutations in cells and fluid of malignant pleural effusion in non-small cell lung cancer. Lung Cancer, 2008, 60: 175-182.
6. Kahn SM, Jiang W, Culbertson TA, et al. Rapid and sensitive nonradioactive detection of mutant K-ras genes via ‘enriched’PCR amplification. Oncogene, 1991 , 6: 1079 -1083 .
7. Chen J, Viola MV. A method to detect ras point mutations in small subpopulations of cells. Anal Biochem, 1991, 195: 51-56.
8. Toyooka S, Tsukuda K, OuchidaM, et al. Detection of codon 61 point mutations of the K-ras gene in lung and colorectal cancers by enriched PCR. Oncol Rep, 2003, 10: 1455-1459.
9. Asano H, Toyooka S, Tokumo M, et al. Detection of EGFR gene mutation in lung cancer by mutant-enriched polymerase chain reaction assay. Clin Cancer Res, 2006, 12: 43-48.
10. Soh J, Toyooka S, Aoe K, et al. Usefulness of EGFR mutation screening in pleural fluid to predict the clinical outcome of gefitinib treated patients with lung cancer. Int J Cancer, 2006 , 119: 2353 - 2358.
11. Kimura H, Fujiwara Y, Sone T, et al. EGFR mutation status in tumour-derived DNA from pleural effusion fluid is a practical basis for predicting the response to gefitinib. Br J Cancer, 2006, 95: 1390- 1395.
12. Kimura H, Fujiwara Y, Sone T, et al. High sensitivity detection of epidermal growth factor receptor mutations in the pleural effusion of non-small cell lung cancer patients. Cancer Sci, 2006, 97: 642-648.
13. Wu SG, Gow CH, Yu CJ, et al. Frequent epidermal growth factor receptor gene mutations in malignant pleural effusion of lung adenocarcinoma. Eur Respir J, 2008, 32: 924-930.
14. Kimura H, Kasahara K, Kawaishi M, et al. Detection of epidermal growth factor receptor mutations in serum as a predictor of the response to gefitinib in patients with non-small-cell lung cancer. Clin Cancer Res, 2006, 12: 3915-3921.
15. Kimura H, Suminoe M, Kasahara K, et al. Evaluation of epidermal growth factor receptor mutation status in serum DNA as a predictor of response to gefitinib( IRESSA) . Br J Cancer, 2007, 97: 778-784.
16. 董強剛, 黃進肅, 韓寶惠, 等. 晚期肺癌患者血清游離DNA 中 EGFR 外顯子19 的基因突變研究. 腫瘤, 2006, 26: 59-63.

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應用酶切富集PCR 法檢測非小細胞肺癌患者胸腔積液表皮生長因子受體基因突變

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應用酶切富集PCR 法檢測非小細胞肺癌患者胸腔積液表皮生長因子受體基因突變

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