Detecting Free Cancer Cells in Peritoneal Cavity of Colorectal Cancer Patients by Branched-Chain DNA and SqRT-PCR_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

WANG Haobin ¹ , WANG Chongshu ¹ , ZHANG Caiquan ² , XIE Xianyong ³

1.The First Department of General Surgery, The Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China;;
2.The Third Department of General Surgery, The First Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400016, China;;
3.Department of Pathology, The Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China;

Corresponding author：

Keywords：

Colorectal cancer; Tumor micrometastasis; Branched-chain DNA; semi-quantitative real time-polymerase chain reaction; Carcinoembryonic antigen

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Abstract Full text Figures/Tables Video References Cited by

Objective To evaluate branched-chain DNA (b-DNA) signal amplification and semi-quantitative (Sq) RT-PCR in detection of free cancer cells in peritoneal flushing fluid of colorectal cancer patients during surgery.
Methods The CEA mRNA in peritoneal flushing fluid in 48 cases of colorectal cancer were detected by b-DNA and SqRT-PCR. Peritoneal flushing fluid cytology (PLC) was conformed simultaneously to detect the free cancer cells. The peritoneal flushing fluid of 12 cases with colorectal benign disease were taken as negative control, GAPDH mRNA as internal control.
Results In colorectal cancer patients, positive rate of free cancer cells by bDNA and SqRT-PCR （43.8%, 31.3%） was higher than that by PLC （4.2%）. The relative quantitative expressions of CEA mRNA were related to the Dukes staging， depth invasion and differentiation degree (P lt;0.05)， but irrelevant to tumor size，the patients’ age and gender (P gt;0.05).
Conclusion Both b-DNA and SqRT-PCR technologies have advantages and disadvantages to detect free cancer cells in peritoneal flushing fluid， which are related to clinicopathological factors.

Citation： WANG Haobin ,WANG Chongshu,ZHANG Caiquan,XIE Xianyong. Detecting Free Cancer Cells in Peritoneal Cavity of Colorectal Cancer Patients by Branched-Chain DNA and SqRT-PCR. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2010, 17(2): 165-169. doi: Copy

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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. 结直肠癌外科应用技术的规范与创新（十五） [J]. 中国普外基础与临床杂志, 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, 周宁新, 高丽杰, 等. 半定量检测胆管癌组织中人fxyd6基因的表达 [J]. 中国普外基础与临床杂志, 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. 骆成玉, 赵丹宁, 李世拥, 等. 大肠癌患者腹腔微量游离癌细胞的研究 [J]. 中华外科杂志, 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.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Detecting Free Cancer Cells in Peritoneal Cavity of Colorectal Cancer Patients by Branched-Chain DNA and SqRT-PCR

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