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http://dx.doi.org/10.5352/JLS.2017.27.11.1290

A PCR Denaturing Gradient Gel Electrophoresis (DGGE) Analysis of Intestinal Microbiota in Gastric Cancer Patients Taking Anticancer Agents  

Yu, Sun Nyoung (Department of Microbiology and Immunology, Pusan National University School of Medicine)
Ahn, Soon Cheol (Department of Microbiology and Immunology, Pusan National University School of Medicine)
Publication Information
Journal of Life Science / v.27, no.11, 2017 , pp. 1290-1298 More about this Journal
Abstract
Intestinal microbiota is an important factor in the development of immune defense mechanisms in the human body. Treatments with anticancer agents, such as 5-Fluorouracil, Cisplatin, and Oxaliplatin, significantly change the temporal stability and environment of intestinal bacterial flora. The anticancer treatment chemotherapy often depresses the immune system and induces side effects, such as diarrhea. This study investigated the effects anticancer agents have on the intestinal microbial ecosystems of patients with gastric cancer. An exploration of the diversity and temporal stability of the dominant bacteria was undertaken using a DGGE with the 16S rDNA gene. Researchers collected stool samples from patients zero, two and eight weeks after the patients started chemotherapy. After the treatment with anticancer agents, the bacteria strains Sphingomonas paucimobilis, Lactobacillus gasseri, Parabacteroides distasonis and Enterobacter sp. increased. This study focused on the survival of the beneficial microorganisms Bifidobacterium and Lactobacillus in the intestines of cancer patients. The administration of antigastric cancer agents significantly decreased Lactobacillus and Bifidobacterium populations and only moderately affected the main bacterial groups in the patients' intestinal ecosystems. The results showed the versatility of a cultivation independent-PCR DGGE analysis regarding the visual monitoring of ecological diversity and anticancer agent-induced changes in patients' complex intestinal microbial ecosystems.
Keywords
Anticancer agent; diversity; intestinal microbiota; PCR-DGGE;
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1 Alvaro, E., Andrieux, C., Rochet, V., Rigottier-Gois, L., Lepercq, P., Sutren, M., Galan, P., Duval, Y., Juste, C. and Dore, J. 2007. Composition and metabolism of the intestinal microbiota in consumers and non-consumers of yogurt. Br. J. Nutr. 97, 126-133.   DOI
2 Barbut, F. and Meynard, J. L. 2002. Managing antibiotic associated diarrhoea. BMJ. 324, 1345-1346.   DOI
3 Dar, S. A., Kuenen, J. G. and Muyzer, G. 2005. Nested PCR-denaturing gradient gel electrophoresis approach to determine the diversity of sulfate-reducing bacteria in complex microbial communities. Appl. Environ. Microbiol. 71, 2325-2330.   DOI
4 Domann, E. 2003. Culture-independent identification of pathogenic bacteria and polymicrobial infections in the genitourinary tract of renal transplant recipients. J. Clin. Microbiol. 41, 5500-5510.   DOI
5 Donskey, C. J., Hujer, A. M., Das, S. M., Pultz, N. J., Bonomo, R. A. and Rice, L. B. 2003. Use of denaturing gradient gel electrophoresis for analysis of the stool microbiota of hospitalized patients. J. Microbiol. Methods 54, 249-256.   DOI
6 Kubota, Y. 1990. Fecal intestinal flora in patients with colon adenoma and colon cancer. Nihon. Shokakibyo. Gakkai. Zasshi. 87, 771-779.
7 Fuentes, S., Egert, M., Jimenez-Valera, M., Ramos-Cormenzana, A., Ruiz-Bravo, A., Smidt, H. and Monteoliva-Sanc hez, M. 2008. Administration of Lactobacillus casei and Lactobacillus plantarum affects the diversity of murine intestinal lactobacilli, but not the overall bacterial community structure. Res. Microbiol. 159, 237-243.   DOI
8 Gillan, D. C. 2004. The effect of an acute copper exposure on the diversity of a microbial community in North Sea sediments as revealed by DGGE analysis--the importance of the protocol. Mar. Pollut. Bull. 49, 504-513.   DOI
9 Kaufmann, P., Pfefferkorn, A., Teuber, M. and Meile, L. 1997. Identification and quantification of Bifidobacterium species isolated from food with genus-specific 16S rRNAtargeted probes by colony hybridization and PCR. Appl. Environ. Microbiol. 63, 1268-1273.
10 Li, M., Gong, J., Cottrill, M., Yu, H., de Lange. C., Burton, J. and Topp, E. 2003. Evaluation of QIAamp DNA Stool Mini Kit for ecological studies of gut microbiota. J. Microbiol. Methods 54, 13-20.   DOI
11 Manninen, T. J., Rinkinen, M. L., Beasley, S. S. and Saris, P. E. 2006. Alteration of the canine small-intestinal lactic acid bacterium microbiota by feeding of potential probiotics. Appl. Environ. Microbiol. 72, 6539-6543.   DOI
12 Martin, R., Heilig, H. G., Zoetendal, E. G., Jimenez, E., Fernandez, L., Smidt, H. and Rodriguez, J. M. 2007. Cultivation-independent assessment of the bacterial diversity of breast milk among healthy women. Res. Microbiol. 158, 31-37.   DOI
13 Picard, C., Fioramonti, J., Francois, A., Robinson, T., Neant, F. and Matuchansky, C. 2005. Review article: bifidobacteria as probiotic agents -- physiological effects and clinical benefits. Aliment Pharmacol. Ther. 22, 495-512.   DOI
14 Matto, J., Maunuksela, L., Kajander K, Palva, A., Korpela, R., Kassinen, A. and Saarela, M. 2005. Composition and temporal stability of gastrointestinal microbiota in irritable bowel syndrome--a longitudinal study in IBS and control subjects. FEMS Immunol. Med. Microbiol. 43, 213-222.   DOI
15 Murray, A. E., Hollibaugh, J. T. and Orrego, C. 1996. Phylogenetic compositions of bacterioplankton from two California estuaries compared by denaturing gradient gel electrophoresis of 16S rDNA fragments. Appl. Environ. Microbiol. 62, 2676-2680.
16 Nielsen, S., Nielsen, D. S., Lauritzen, L., Jakobsen, M. and Michaelsen, K. F. 2007. Impact of diet on the intestinal microbiota in 10-month-old infants. J. Pediatr. Gastroenterol Nutr. 44, 613-618.   DOI
17 Saxelin, M., Tynkkynen, S., Mattila-Sandholm, T. and de Vos, W. M. 2005. Probiotic and other functional microbes: from markets to mechanisms. Curr. Opin. Biotechnol. 16, 204-211.   DOI
18 Singh, J., Rivenson, A., Tomita, M., Shimamura, S., Ishibashi, N. and Reddy, B. S. 1997. Bifidobacterium longum, a lactic acid-producing intestinal bacterium inhibits colon cancer and modulates the intermediate biomarkers of colon carcinogenesis. Carcinogenesis 18, 833-841.   DOI
19 Smalla, K. 2007. Bacterial diversity of soils assessed by DGGE, T-RFLP and SSCP fingerprints of PCR-amplified 16S rRNA gene fragments: do the different methods provide similar results? J. Microbiol. Methods 69, 470-479.   DOI
20 Stebbings, S., Munro, K., Simon, M. A., Tannock, G., Highton, J., Harmsen, H., Welling, G., Seksik, P., Dore, J., Grame, G. and Tilsala-Timisjarvi, A. 2002 Comparison of the faecal microflora of patients with ankylosing spondylitis and controls using molecular methods of analysis. Rheumatology 41, 1395-1401.   DOI
21 Thompson-Chagoyan, O. C., Maldonado, J. and Gil, A. 2007. Colonization and impact of disease and other factors on intestinal microbiota. Dig. Dis. Sci. 52, 2069-2077.   DOI
22 Vannucci, L., Stepankova, R., Kozakova, H., Fiserova, A., Rossmann, P. and Tlaskalova-Hogenova, H. 2008. Colorectal carcinogenesis in germ-free and conventionally reared rats: different intestinal environments affect the systemic immunity. Int. J. Oncol. 32, 609-617.
23 Wang, M., Ahrne, S., Jeppsson, B. and Molin, G. 2005. Comparison of bacterial diversity along the human intestinal tract by direct cloning and sequencing of 16S rRNA genes. FEMS Microbiol. Ecol. 54, 219-231.   DOI