SYBR GreenⅠ Real-Time Fluorescence Quantitative PCR Analysis of Variation of Intestinal Microflora in Patients with Colorectal Cancer_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

GUO Shikui ¹ , BAO Weimin ¹ , GONG Kunmei ¹ , SHAO Jianchun ² , CHEN Di ² , WANG Kunhua ¹

1.The First Department of General Surgery, The Kunhua Affiliated Hospital of Kunming Medical College (The First People’s Hospital of Yunnan Province), Kunming 650032, China;;
2.Department of Clinical Laboratory, The First People’s Hospital of Kunming, Kunming 650011, China;

Corresponding author：

Keywords：

Colorectal cancer; Intestinal microflora; Real-time fluorescence quantitative PCR; SYBR Green Ⅰ

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To analyze the variation of intestinal microflora in patients with colorectal cancer by SYBR GreenⅠreal-time fluorescence quantitative PCR and reveal the role and significance of intestinal microflora in the colorectal cancer-associated molecular pathogenesis.
Methods A set of 16S rRNA gene group of species-specific primers for Bifidobacterium spp., Lactobacillus group, Escherichia coli, and ddl gene-targeted species-specific primers for Enterococcus faecalis and feces Enterococcus were designed. Patients with colorectal cancer (colorectal cancer group, n=30) and healthy volunteers (normal control group, n=30) were included and whose feces were collected to extract bacterial genome DNA. SYBR GreenⅠ real-time fluorescence quantitative PCR was used to analyze the five mentioned bacterial amounts.
Results Level of Bifidobacterium spp. (4.52±0.49) and Lactobacillus group (5.46±0.12) in colorectal cancer group were significantly lower than those （9.25±0.83 and 7.45±0.37） of normal control group (P lt;0.05), whereas levels of Escherichia coli (5.82±0.47), Enterococcus faecalis (10.6±0.30) and feces Enterococcus (5.74±0.16) in colorectal cancer group were significantly higher than those （4.68±0.32, 4.95±0.24, and 5.03±0.43） of normal control group (P lt;0.05).
Conclusions The fecal microflora composition of patients with colorectal cancer is significantly decreased in Bifidobacterium spp. and Lactobacillus group, whereas increased in Escherichia coli, Enterococcus faecalis, and feces Enterococcus. These data underline that the occurrence and progress of colorectal cancer may be related to intestinal microflora.

Citation： GUO Shikui ,BAO Weimin,GONG Kunmei,SHAO Jianchun,CHEN Di,WANG Kunhua. SYBR GreenⅠ Real-Time Fluorescence Quantitative PCR Analysis of Variation of Intestinal Microflora in Patients with Colorectal Cancer. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2010, 17(5): 463-468. doi: Copy

1.	Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics, 2002 [J]. CA Cancer J Clin, 2005; 55(2): 74-108.
2.	徐富星．大肠癌研究现状 [J]．国际消化病杂志， 2006； 26(6)： 365-366, 424．.
3.	Sung JJ, Lau JY, Goh KL, et al. Increasing incidence of colorectal cancer in Asia: implications for screening [J]. Lancet Oncol, 2005; 6(11): 871-876.
4.	Winawer SJ. Screening of colorectal cancer: progress and problems [J]． Recent Results Cancer Res, 2005; 166: 231-244.
5.	朱宝成，王磊. 大肠癌筛查技术的研究进展 [J]. 吉林医学, 2009； 30(5)： 459-462.
6.	白鹏，吕愈敏，顾芳. 细菌16S rDNA荧光定量PCR法分析溃疡性结肠炎患者肠道菌群变化 [J]. 胃肠病学和肝病学杂志， 2008； 17(7)： 566-571.
7.	Guarner F. Enteric flora in health and disease [J]. Digestion, 2006; 73 Suppl 1: 5-12.
8.	McGarr SE, Ridlon JM, Hylemon PB. Diet, anaerobic bacterial metabolism, and colon cancer: a review of the literature [J]. J Clin Gastroenterol, 2005; 39(2): 98-109.
9.	任月，袁杰利，卢行安，等. 粪便中肠球菌SYBR GreenⅠ荧光定量PCR检测方法的建立 [J]. 中国微生态学杂志， 2007; 19(6): 499-501.
10.	Byun R, Nadkarni MA, Chhour KL, et al. Quantitative analysis of diverse Lactobacillus species present in advanced dental caries [J]. J Clin Microbiol, 2004; 42(7): 3128-3136.
11.	Wong ML, Medrano JF. Real-time PCR for mRNA quantit-ation [J]. Biotechniques, 2005; 39(1): 75-85．.
12.	范莹，吴硕东，于宏，等. 腹腔镜结直肠癌根治术与同期开腹手术对比的单中心回顾性研究 [J]. 中国普外基础与临床杂志， 2009； 16(5)： 384-388.
13.	刘国伟，蔡全才. 大肠癌筛查现状及早期诊断进展 [J]. 现代肿瘤医学， 2009； 17(4)： 770-774.
14.	张洪伟，同李平，孙力. 结直肠癌的个体化治疗进展 [J]. 中国普外基础与临床杂志， 2009； 16(9)： 689-692.
15.	郑树．结直肠癌的预防 [J]．中华肿瘤防治杂志， 2006； 13(1)： 1-2.
16.	郑树．我国的结直肠癌筛查方案 [J]．医学研究杂志， 2006； 35(2)： 8-9．.
17.	Sung J. Does fecal occult blood test have a place for colorectal cancer screening in China in 2006? [J]. Am J Gastroenterol, 2006; 101(2): 213-215.
18.	Ravva SV, Stanker LH. Real-time quantitative PCR detection of Mycobacterium avium subsp. paratuberculosis and differentiation from other mycobacteria using SYBR Green and TaqMan assays [J]. J Microbiol Methods, 2005; 63(3): 305-317.
19.	Sghir A, Gramet G, Suau A, et al． Quantification of bacterial groups within human fecal flora by oligonucleotide probe hybridization [J]． Appl Environ Microbiol, 2000; 66(5): 2263-2266．.
20.	Guarner F, Malagelada JR． Gut flora in health and disease, [J]． Lancet, 2003; 361(9356): 512-519.
21.	Ott SJ, Musfeldt M, Ullmann U, et al． Quantification of intestinal bacterial populations by real-time PCR with a universal primer set and minor groove binder probes: a global approach to the enteric flora [J]． J Clin Microbiol, 2004; 42(6): 2566-2572.
22.	张纪伟，杜鹏，杨宝仁，等. 补充双歧三联活菌对大肠癌术后肠道菌群的影响 [J]. 上海交通大学学报(医学版), 2008；28(10): 1285-1287, 1300.
23.	Rafter J, Bennett M, Caderni G, et al. Dietary synbiotics reduce cancer risk factors in polypectomized and colon cancer patients [J]． Am J Clin Nutr, 2007; 85(2): 488-496.
24.	包维民，王昆华，唐映梅，等. 结直肠癌患者手术前后肠道菌群与正常人群的比较 [J]. 结直肠肛门外科， 2009； 15(4): 224-226.
25.	Iwashita J, Sato Y, Sugaya H, et al．mRNA of MUC2 is stimulated by IL-4, IL-13 or TNF-alpha through a mitogen-activated protein kinase pathway in human colon cancer cells [J]. Immunol Cell Biol, 2003; 81(4): 275-282.
26.	Rescigno M. The pathogenic role of intestinal flora in IBD and colon cancer [J]. Curr Drug Targets, 2008; 9(5): 395-403.

1. Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics, 2002 [J]. CA Cancer J Clin, 2005; 55(2): 74-108.
2. 徐富星．大肠癌研究现状 [J]．国际消化病杂志， 2006； 26(6)： 365-366, 424．.
3. Sung JJ, Lau JY, Goh KL, et al. Increasing incidence of colorectal cancer in Asia: implications for screening [J]. Lancet Oncol, 2005; 6(11): 871-876.
4. Winawer SJ. Screening of colorectal cancer: progress and problems [J]． Recent Results Cancer Res, 2005; 166: 231-244.
5. 朱宝成，王磊. 大肠癌筛查技术的研究进展 [J]. 吉林医学, 2009； 30(5)： 459-462.
6. 白鹏，吕愈敏，顾芳. 细菌16S rDNA荧光定量PCR法分析溃疡性结肠炎患者肠道菌群变化 [J]. 胃肠病学和肝病学杂志， 2008； 17(7)： 566-571.
7. Guarner F. Enteric flora in health and disease [J]. Digestion, 2006; 73 Suppl 1: 5-12.
8. McGarr SE, Ridlon JM, Hylemon PB. Diet, anaerobic bacterial metabolism, and colon cancer: a review of the literature [J]. J Clin Gastroenterol, 2005; 39(2): 98-109.
9. 任月，袁杰利，卢行安，等. 粪便中肠球菌SYBR GreenⅠ荧光定量PCR检测方法的建立 [J]. 中国微生态学杂志， 2007; 19(6): 499-501.
10. Byun R, Nadkarni MA, Chhour KL, et al. Quantitative analysis of diverse Lactobacillus species present in advanced dental caries [J]. J Clin Microbiol, 2004; 42(7): 3128-3136.
11. Wong ML, Medrano JF. Real-time PCR for mRNA quantit-ation [J]. Biotechniques, 2005; 39(1): 75-85．.
12. 范莹，吴硕东，于宏，等. 腹腔镜结直肠癌根治术与同期开腹手术对比的单中心回顾性研究 [J]. 中国普外基础与临床杂志， 2009； 16(5)： 384-388.
13. 刘国伟，蔡全才. 大肠癌筛查现状及早期诊断进展 [J]. 现代肿瘤医学， 2009； 17(4)： 770-774.
14. 张洪伟，同李平，孙力. 结直肠癌的个体化治疗进展 [J]. 中国普外基础与临床杂志， 2009； 16(9)： 689-692.
15. 郑树．结直肠癌的预防 [J]．中华肿瘤防治杂志， 2006； 13(1)： 1-2.
16. 郑树．我国的结直肠癌筛查方案 [J]．医学研究杂志， 2006； 35(2)： 8-9．.
17. Sung J. Does fecal occult blood test have a place for colorectal cancer screening in China in 2006? [J]. Am J Gastroenterol, 2006; 101(2): 213-215.
18. Ravva SV, Stanker LH. Real-time quantitative PCR detection of Mycobacterium avium subsp. paratuberculosis and differentiation from other mycobacteria using SYBR Green and TaqMan assays [J]. J Microbiol Methods, 2005; 63(3): 305-317.
19. Sghir A, Gramet G, Suau A, et al． Quantification of bacterial groups within human fecal flora by oligonucleotide probe hybridization [J]． Appl Environ Microbiol, 2000; 66(5): 2263-2266．.
20. Guarner F, Malagelada JR． Gut flora in health and disease, [J]． Lancet, 2003; 361(9356): 512-519.
21. Ott SJ, Musfeldt M, Ullmann U, et al． Quantification of intestinal bacterial populations by real-time PCR with a universal primer set and minor groove binder probes: a global approach to the enteric flora [J]． J Clin Microbiol, 2004; 42(6): 2566-2572.
22. 张纪伟，杜鹏，杨宝仁，等. 补充双歧三联活菌对大肠癌术后肠道菌群的影响 [J]. 上海交通大学学报(医学版), 2008；28(10): 1285-1287, 1300.
23. Rafter J, Bennett M, Caderni G, et al. Dietary synbiotics reduce cancer risk factors in polypectomized and colon cancer patients [J]． Am J Clin Nutr, 2007; 85(2): 488-496.
24. 包维民，王昆华，唐映梅，等. 结直肠癌患者手术前后肠道菌群与正常人群的比较 [J]. 结直肠肛门外科， 2009； 15(4): 224-226.
25. Iwashita J, Sato Y, Sugaya H, et al．mRNA of MUC2 is stimulated by IL-4, IL-13 or TNF-alpha through a mitogen-activated protein kinase pathway in human colon cancer cells [J]. Immunol Cell Biol, 2003; 81(4): 275-282.
26. Rescigno M. The pathogenic role of intestinal flora in IBD and colon cancer [J]. Curr Drug Targets, 2008; 9(5): 395-403.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

SYBR GreenⅠ Real-Time Fluorescence Quantitative PCR Analysis of Variation of Intestinal Microflora in Patients with Colorectal Cancer

Abstract Full text Figures/Tables Video References Cited by

Format

Content