• 1.川北醫(yī)學(xué)院附屬醫(yī)院普外一科（四川南充 637000）;;
  • 2.重慶醫(yī)科大學(xué)附屬第一醫(yī)院普外三科（重慶 400016）;;
  • 3.川北醫(yī)學(xué)院附屬醫(yī)院病理科（四川南充 637000）;

目的  探討分支DNA（b-DNA）及半定量RT-PCR（SqRT-PCR)在結(jié)直腸癌術(shù)中腹腔沖洗液中游離癌細(xì)胞檢測中的應(yīng)用。
方法  分別采用基于b-DNA信號(hào)放大的基因表達(dá)定量檢測技術(shù)及SqRT-PCR方法檢測48例結(jié)直腸癌患者術(shù)中腹腔沖洗液中CEA mRNA的表達(dá)，同時(shí)行腹腔沖洗液細(xì)胞學(xué)檢查（peritoneal lavage cytology，PLC）,收集12例結(jié)直腸良性病變患者的腹腔沖洗液為陰性對(duì)照，GAPDH mRNA為內(nèi)參對(duì)照。
結(jié)果  b-DNA技術(shù)和SqRT-PCR方法檢測游離癌細(xì)胞的陽性率（43.8%，31.3%）較PLC的檢出率（4.2%）高（P lt;0.01）。結(jié)直腸癌患者腹腔沖洗液中CEA mRNA的相對(duì)表達(dá)量均與腫瘤分化程度、漿膜侵犯程度和Dukes分期有關(guān)（P lt;0.05），而與腫瘤大小、患者性別、年齡無關(guān)（P gt;0.05）。
結(jié)論  b-DNA技術(shù)和SqRT-PCR方法檢測游離癌細(xì)胞各有優(yōu)缺點(diǎn); 腹腔內(nèi)游離癌細(xì)胞的存在與結(jié)直腸癌臨床病理因素有關(guān)。

引用本文： 王浩斌,王崇樹,張才全,謝賢雍. 應(yīng)用分支DNA 技術(shù)及SqRT-PCR方法檢測結(jié)直腸癌患者腹腔沖洗液中游離癌細(xì)胞. 中國普外基礎(chǔ)與臨床雜志, 2010, 17(2): 165-169. doi: 復(fù)制

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3. Barber RD, Harmer DW, Coleman RA, et al. GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues [J]. Physiol Genomics, 2005; 21(3): 389395.
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13. 駱成玉, 趙丹寧, 李世擁, 等. 大腸癌患者腹腔微量游離癌細(xì)胞的研究 [J]. 中華外科雜志, 2001; 39（6）: 433436.
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15. Knudsen BS, Allen AN, Mclerran DF, et al. Evalluation of branchedchain DNA assay for measurement of RNA in formalinfoxed tissures [J]. J Mol Diaqn, 2008; 10(2): 169176.
16. Berrodin TJ, Jelinsky SA, Graciani N, et al. Novel progesterone receptor modulators with gene selective and contextdependent partial agonism [J]. Biochem Pharmacol, 2009; 77(2): 204215.
17. Lee AC, Dai Z, Chen B, et al. Electrochemical branchedDNA assay for polymerase chain reactionfree detection and quantification of oncogenes in messenger RNA [J]. Anal Chem, 2008; 80(24): 94029410.
  1. 1. Akasu T， Sugihara K. Solitary peritoneal metastasis from colon carcinoma [J]. Surgery, 1998; 124(5): 938940.
  2. 2. Suzuki T, Higgins PJ, Crawford DR. Control selection for RNA quantitation [J]. Biotechniques, 2000; 29(2): 332337.
  3. 3. Barber RD, Harmer DW, Coleman RA, et al. GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues [J]. Physiol Genomics, 2005; 21(3): 389395.
  4. 4. 李立. 結(jié)直腸癌外科應(yīng)用技術(shù)的規(guī)范與創(chuàng)新（十五） [J]. 中國普外基礎(chǔ)與臨床雜志, 2008; 15(3): 215217.
  5. 5. Yamamoto S, Akasu T, Fujita S, et al. Longterm prognostic value of conventional peritoneal cytology after curative resection for colorectal carcinoma [J]. Jpn J Clin Oncol, 2003; 33(1): 3337.
  6. 6. Noura S, Ohue M, Seki Y, et al. Longterm prognostic value of conventional peritoneal lavage cytology in patients undergoing curative colorectal cancer resection [J]. Dis Colon Rectum, 2009; 52(7): 13121320.
  7. 7. Japink D, Leers MP, Sosef MN, et al. CEA in activated macrophages. New diagnostic possibilities for tumor markers in early colorectal cancer [J]. Anticancer Res, 2009; 29(8): 32453251.
  8. 8. Prochotsky A, Okolicany R, Sekac J, et al. Diagnosis and management of local and locoregional recurrence of colorectal carcinoma [J]. Bratisl Lek Listy, 2009; 110(9): 569573.
  9. 9. Katoh H, Yamashita K, Sato T, et al. Prognostic significance of peritoneal tumour cells identified at surgery for colorectal cancer [J]. Br J Surg, 2009; 96(7): 769777.
  10. 10. 肖梅, 周寧新, 高麗杰, 等. 半定量檢測膽管癌組織中人fxyd6基因的表達(dá) [J]. 中國普外基礎(chǔ)與臨床雜志, 2008; 15(2): 8387.
  11. 11. Freeman WM, Walker SJ, Vrana KE. Quantitative RTPCR: pitfalls and potential [J]. Biotechniques, 1999; 26(1): 112122.
  12. 12. Nakanishi H, Kodera Y, Yamamura Y, et al. Rapid quantitative detection of carcinoembryonic antigenexpressing free tumor cells in the peritoneal cavity of gastriccancer patients with realtime RTPCR on the lightcycler [J]. Int J Cancer, 2000; 89(5): 411417.
  13. 13. 駱成玉, 趙丹寧, 李世擁, 等. 大腸癌患者腹腔微量游離癌細(xì)胞的研究 [J]. 中華外科雜志, 2001; 39（6）: 433436.
  14. 14. Canales, RD, Luo Y, Willey JC, et al. Evaluation of DNA microarray results with quantitative gene expression platforms [J].Nat Biotechnol, 2006; 24(9): 11151122.
  15. 15. Knudsen BS, Allen AN, Mclerran DF, et al. Evalluation of branchedchain DNA assay for measurement of RNA in formalinfoxed tissures [J]. J Mol Diaqn, 2008; 10(2): 169176.
  16. 16. Berrodin TJ, Jelinsky SA, Graciani N, et al. Novel progesterone receptor modulators with gene selective and contextdependent partial agonism [J]. Biochem Pharmacol, 2009; 77(2): 204215.
  17. 17. Lee AC, Dai Z, Chen B, et al. Electrochemical branchedDNA assay for polymerase chain reactionfree detection and quantification of oncogenes in messenger RNA [J]. Anal Chem, 2008; 80(24): 94029410.