Comparative Study on Detection of Peritoneal Free Cancer Cells of Gastric Surgery_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LIU Zhenfang , HAN Xiaopeng , LIU Hongbin , ZHU Wankun , SU Lin.

Department of General Surgery, Lanzhou General Hospital, Lanzhou Command PLA, Lanzhou 730050, China;

Corresponding author：

Keywords：

Gastric cancer; Peritoneal lavage fluid; Cytologic examination; Flow cytometry; Realtime fluorescent quantitative reverse transcriptionpolymerase chain reaction

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Objective To investigate the detection of peritoneal free cancer cells and its clinical significance.
Methods The peritoneal free cancer cells, the positive rates of CK20 protein and CK20 mRNA expressions of peritoneal lavage fluid were detected by peritoneal lavage cytology (PLC), flow cytometry (FCM) and real-time fluorescent quantitative RT-PCR in 50 cases of gastric cancer patients, respectively. The sensitivity of three kinds of detection method to peritoneal free cancer cells was compared.
Results The positive rates of peritoneal free cancer cells, CK20 protein and mRNA expression of peritoneal lavage fluid were 20.0% (10/50), 36.0% (18/50) and 58.0% (29/50), respectively. The positive rate of CK20 mRNA expression detected by real-time fluorescencequantitative RT-PCR in peritoneal lavage fluid was significantly higher than those of the CK20 protein expression detected by FCM and peritoneal free cancer cells detected by PLC (P lt;0.05 or P lt;0.001). The difference of positive rate of CK20 protein expression and peritoneal free cancer cells was not significant (P gt;0.05). The positive rate of CK20 mRNA expression of peritoneal lavage fluid was related to the tumor invasion depth, differentiation degree, TNM stage, and lymph node metastasis (P lt;0.05). Conclusion Real-time fluorescence quantitative RT-PCR is an effective method for the detection of peritoneal free cancer cells.

Citation： LIU Zhenfang,HAN Xiaopeng,LIU Hongbin,ZHU Wankun,SU Lin.. Comparative Study on Detection of Peritoneal Free Cancer Cells of Gastric Surgery. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2010, 17(8): 803-807. doi: Copy

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13.	李乐, 伍严安, 黄源茂. 胸腹水脱落细胞流式细胞法DNA倍体分析的临床诊断价值 [J]. 福建医科大学学报, 2007; 41(2): 159162.
14.	Mitas M, Mikhitarian K, Walters C, et al. Quantitative realtime RTPCR detection of breast cancer micrometastasis using a multigene marker panel [J]. Int J Cancer, 2001; 93(2): 162171.
15.	Bustin SA. Absolute quantification of mRNA using realtime reverse transcription polymerase chain reaction assays [J]. J Mol Endocrinol, 2000; 25(2): 169193.
16.	Klein D. Quantification using realtime PCR technology: applications and limitations [J]. Trends Mol Med, 2002; 8(6): 257260.
17.	Chen XM, Chen GY, Wang ZR, et al. Detection of micrometastasis of gastric carcinoma in peripheral blood circulation [J]. World J Gastroenterol, 2004; 10(6): 804808.
18.	熊兵红, 程勇, 王严庆. 细胞角蛋白 20检测在大肠癌微转移中的临床意义 [J]. 世界华人消化杂志, 2006; 14(14): 13941402.
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20.	Osaka H, Yashiro M, Sawada T, et al. Is a lymph node detected by the dyeguided method a true sentinel node in gastric cancer? [J]. Clin Cancer Res, 2004; 10(20): 69126918.
21.	叶凯, 许建华, 郭启祥. 胃癌腹腔微转移的检测及其预后意义 [J]. 实用癌症杂志, 2004; 19(4): 415417.

1. Yokota T, Ishiyama S, Saito T, et al. Lymph node metastasis as a significant prognosis factor in gastric cancer: a multiple logistic regression analysis [J]. Scand J Gastroenterol, 2004; 39(4): 380384.
2. 王舒宝, 王俊. 胃癌复发与转移的有关问题及综合治疗 [J]. 中国普外基础与临床杂志, 2006; 13(1): 911.
3. Fleming ID, Cooper JS, Henson DE, et al. American Joint Committee on Cancer Staging Manual [M]. 5th ed. Philadelphia: LippincottRaven, 1997: 71.
4. Crew KD, Neugut AI. Epidemiology of gastric cancer [J]. World J Gastroenterol, 2006; 12(3): 354362.
5. Lee CC, Lo SS, Wu CW, et al. Peritoneal recurrence of gastric adenocarcinoma after curative resection [J]. Hepatogastroenterology, 2003; 50(53): 17201722.
6. McCulloch P. The role of surgery in patients with advanced gastric cancer [J]. Best Pract Res Clin Gastroenterol, 2006; 20(4): 767787.
7. Oyama K, Terashima M, Takagane A, et al. Prognostic significance of peritoneal minimal residual disease in gastric cancer detected by reverse transcriptionpolymerase chain reaction [J]. Br J Surg, 2004; 91(4): 435443.
8. 王昭, 詹文华, 何裕隆, 等. 胃癌患者N2淋巴结转移和腹膜扩散的相关因素和预后分析 [J]. 中国普通外科杂志, 2006; 15(9): 645649.
9. Hohenberger P, Gretschel S. Gastric cancer [J]. Lancet, 2003; 362(9380): 305315.
10. 达明绪, 赵紫罡, 罗婷, 等. 胃癌细胞增殖与腹腔灌洗液中端粒酶活性及腹膜转移的相关性研究 [J]. 中国普外基础与临床杂志, 2006; 13(1): 2528.
11. Yamamoto M, Matsuyama A, Kameyama T, et al. Prognostic reevaluation of peritoneal lavage cytology in Japanese patients with gastric carcinoma [J]. Hepatogastroenterology, 2009; 56(89): 261265.
12. Homma Y, Ushida S, Yamada M, et al. Positive peritoneal washing cytology in multiple cavities can predict poor prognosis of advanced gastric cancer patients [J]. Ann Surg Oncol, 2010; 17(2): 455460.
13. 李乐, 伍严安, 黄源茂. 胸腹水脱落细胞流式细胞法DNA倍体分析的临床诊断价值 [J]. 福建医科大学学报, 2007; 41(2): 159162.
14. Mitas M, Mikhitarian K, Walters C, et al. Quantitative realtime RTPCR detection of breast cancer micrometastasis using a multigene marker panel [J]. Int J Cancer, 2001; 93(2): 162171.
15. Bustin SA. Absolute quantification of mRNA using realtime reverse transcription polymerase chain reaction assays [J]. J Mol Endocrinol, 2000; 25(2): 169193.
16. Klein D. Quantification using realtime PCR technology: applications and limitations [J]. Trends Mol Med, 2002; 8(6): 257260.
17. Chen XM, Chen GY, Wang ZR, et al. Detection of micrometastasis of gastric carcinoma in peripheral blood circulation [J]. World J Gastroenterol, 2004; 10(6): 804808.
18. 熊兵红, 程勇, 王严庆. 细胞角蛋白 20检测在大肠癌微转移中的临床意义 [J]. 世界华人消化杂志, 2006; 14(14): 13941402.
19. Mori K, Aoyagi K, Ueda T, et al. Highly specific marker genes for detecting minimal gastric cancer cells in cytology negative peritoneal washings [J]. Biochem Biophys Res Commun, 2004; 313(4): 931937.
20. Osaka H, Yashiro M, Sawada T, et al. Is a lymph node detected by the dyeguided method a true sentinel node in gastric cancer? [J]. Clin Cancer Res, 2004; 10(20): 69126918.
21. 叶凯, 许建华, 郭启祥. 胃癌腹腔微转移的检测及其预后意义 [J]. 实用癌症杂志, 2004; 19(4): 415417.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Comparative Study on Detection of Peritoneal Free Cancer Cells of Gastric Surgery

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