• 廣州醫(yī)學(xué)院第一附屬醫(yī)院廣州呼吸疾病研究所 呼吸病國(guó)家重點(diǎn)實(shí)驗(yàn)室（廣東廣州 510120）;

目的 探討酶切富集PCR 法檢測(cè)非小細(xì)胞肺癌( NSCLC) 患者胸腔積液表皮生長(zhǎng)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(duì)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(jìn)行分析。結(jié)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(tǒng)計(jì)學(xué)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(jié)論 酶切富集PCR 法可以高效、經(jīng)濟(jì)、準(zhǔn)確地檢測(cè)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(yù)測(cè)方法。

引用本文： 何臣,劉明,徐軍. 應(yīng)用酶切富集PCR 法檢測(cè)非小細(xì)胞肺癌患者胸腔積液表皮生長(zhǎng)因子受體基因突變. 中國(guó)呼吸與危重監(jiān)護(hù)雜志, 2009, 09(5): 451-455. doi: 復(fù)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(qiáng)剛, 黃進(jìn)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. 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. 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. 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. 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. 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. 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. 7. Chen J, Viola MV. A method to detect ras point mutations in small subpopulations of cells. Anal Biochem, 1991, 195: 51-56.
  8. 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. 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. 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. 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. 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. 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. 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. 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. 16. 董強(qiáng)剛, 黃進(jìn)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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