應用分支DNA 技術及SqRT-PCR方法檢測結直腸癌患者腹腔沖洗液中游離癌細胞_《中國普外基礎與臨床雜志》

作者：

王浩斌 ¹ , 王崇樹 ¹ , 張才全 ² , 謝賢雍 ³

1.川北醫學院附屬醫院普外一科（四川南充 637000）;;
2.重慶醫科大學附屬第一醫院普外三科（重慶 400016）;;
3.川北醫學院附屬醫院病理科（四川南充 637000）;

關鍵詞：

結直腸腫瘤腫瘤微轉移分支DNA 半定量反轉錄-聚合酶鏈反應癌胚抗原

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目的探討分支DNA（b-DNA）及半定量RT-PCR（SqRT-PCR)在結直腸癌術中腹腔沖洗液中游離癌細胞檢測中的應用。
方法分別采用基于b-DNA信號放大的基因表達定量檢測技術及SqRT-PCR方法檢測48例結直腸癌患者術中腹腔沖洗液中CEA mRNA的表達，同時行腹腔沖洗液細胞學檢查（peritoneal lavage cytology，PLC）,收集12例結直腸良性病變患者的腹腔沖洗液為陰性對照，GAPDH mRNA為內參對照。
結果 b-DNA技術和SqRT-PCR方法檢測游離癌細胞的陽性率（43.8%，31.3%）較PLC的檢出率（4.2%）高（P lt;0.01）。結直腸癌患者腹腔沖洗液中CEA mRNA的相對表達量均與腫瘤分化程度、漿膜侵犯程度和Dukes分期有關（P lt;0.05），而與腫瘤大小、患者性別、年齡無關（P gt;0.05）。
結論 b-DNA技術和SqRT-PCR方法檢測游離癌細胞各有優缺點; 腹腔內游離癌細胞的存在與結直腸癌臨床病理因素有關。

引用本文： 王浩斌,王崇樹,張才全,謝賢雍. 應用分支DNA 技術及SqRT-PCR方法檢測結直腸癌患者腹腔沖洗液中游離癌細胞. 中國普外基礎與臨床雜志, 2010, 17(2): 165-169. doi: 復制

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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胞的研究 [J]. 中華外科雜志, 2001; 39（6）: 433436.
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.	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.
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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. Akasu T， Sugihara K. Solitary peritoneal metastasis from colon carcinoma [J]. Surgery, 1998; 124(5): 938940.
2. Suzuki T, Higgins PJ, Crawford DR. Control selection for RNA quantitation [J]. Biotechniques, 2000; 29(2): 332337.
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. 李立. 結直腸癌外科應用技術的規范與創新（十五） [J]. 中國普外基礎與臨床雜志, 2008; 15(3): 215217.
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. 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. 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. 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. 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. 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 [J]. 中國普外基礎與臨床雜志, 2008; 15(2): 8387.
11. Freeman WM, Walker SJ, Vrana KE. Quantitative RTPCR: pitfalls and potential [J]. Biotechniques, 1999; 26(1): 112122.
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. 駱成玉, 趙丹寧, 李世擁, 等. 大腸癌患者腹腔微量游離癌細胞的研究 [J]. 中華外科雜志, 2001; 39（6）: 433436.
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. 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.

《中國普外基礎與臨床雜志》

應用分支DNA 技術及SqRT-PCR方法檢測結直腸癌患者腹腔沖洗液中游離癌細胞

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《中國普外基礎與臨床雜志》

應用分支DNA 技術及SqRT-PCR方法檢測結直腸癌患者腹腔沖洗液中游離癌細胞

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

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