Antiviral activity of Bifidobacterium adolescentis SPM1005-A on human papillomavirus type 16

BMC Medicine

Nam-Joo Ha³  Kyung-Jae Kim³  Jeong-Hyun Kwon²  Seon-Hee Shin¹  Si-Won Lee³  Hyang-Mi An³  Do-Kyung Lee³  Min-Kyeong Cha³ 

[1] Department of Pediatrics, College of Medicine, Hallym University, Chuncheon, Republic of Korea;Department of Sport Medicine, Jung Won University, Chungbuk, Republic of Korea;College of Pharmacy, Sahmyook University, Seoul, Republic of Korea

关键词: quantitative real-time PCR (qRT-PCR);    human papillomavirus (HPV) type 16;    Bifidobacterium adolescentis SPM1005-A;    antiviral activity;   

Others  :  1125989
DOI  :  10.1186/1741-7015-10-72

 received in 2012-04-20, accepted in 2012-07-12,  发布年份 2012

【 摘 要 】

Background

Probiotic lactic acid bacteria (LAB) support a functional and balanced immune system, and contribute to immune modulatory effects in combatting microbial pathogens, including viruses. Most cervical cancers are associated with anogenital region infection with high-risk (HR) human papillomavirus (HPV). In this study, we analyzed the antiviral activity of Bifidobacterium adolescentis SPM1005-A in the SiHa cervical cancer cell line expressing HPV type 16.

Methods

We assessed the cellular toxicity of B. adolescentis SPM1005-A in SiHa cells by the Trypan blue dye exclusion assay. Cells (3.6 × 10⁵) in culture plates with or without B. adolescentis SPM1005-A in the same type of medium, were incubated with HPV type 16 at a concentration of 5.1 × 10⁷ cfu/ml. For antiviral analysis, we performed quantitative real-time PCR (qRT-PCR) for E6 and E7 oncogene expressions and observed protein levels by immunoblotting.

Results

The qRT-PCR results showed that E6 and E7 mRNA levels decreased simultaneously. Western blot analysis revealed that the E6 protein expression slightly decreased after 24 and 48 h, but the level of E7 protein expression appear unaffected compared with that in the control. Decreased HPV16 E6 and E7 mRNA transcript and protein levels were not associated with cell morphology or significant cytotoxic effects.

Conclusions

This study showed that B. adolescentis SPM1005-A had antiviral activity through suppression E6 and E7 oncogene expression. The results suggest that B. adolescentis SPM1005-A could be potential applications of HPV-associated cervical cancer prevention.

【 授权许可】

   
2012 Cha et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Arbyn M, Castellsagué X, de Sanjosé S, Bruni L, Saraiya M, Bray F, Ferlay J: Worldwide burden of cervical cancer in 2008. Ann Oncol 2011, 22:2675-2686.
• [2]Campisi G, Panzarella V, Giuliani M, Lajolo C, Di Fede O, Falaschini S, Di Liberto C, Scully C, Lo Muzio L: Human papillomavirus: its identity and controversial role in oral oncogenesis, premalignant and malignant lesions. Int J Oncol 2007, 30:813-823.
• [3]Münger K, Baldwin A, Edwards KM, Hayakawa H, Nguyen CL, Owens M, Grace M, Huh K: Mechanisms of human papillomavirus-induced oncogenesis. J Virol 2004, 78:11451-11460.
• [4]Walboomers JM, Jacobs MV, Manos MM, Bosch FX, Kummer JA, Shah KV, Snijders PJ, Peto J, Meijer CJ, Muñoz N: Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 1999, 189:12-19.
• [5]Um SJ, Lee SY, Kim EJ, Myoung J, Namkoong SE, Park JS: Down-regulation of human papillomavirus E6/E7 oncogene by arsenic trioxide in cervical carcinoma cells. Cancer Lett 2002, 181:11-22.
• [6]Thomas M, Pim D, Banks L: The role of the E6-p53 interaction in the molecular pathogenesis of HPV. Oncogene 1999, 18:7690-7700.
• [7]Huh KW, DeMasi J, Ogawa H, Nakatani Y, Howley PM, Münger K: Association of the human papillomavirus type 16 E7 oncoprotein with the 600-kDa retinoblastoma protein-associated factor, p600. Proc Natl Acad Sci USA 2005, 102:11492-11497.
• [8]Huh K, Zhou X, Hayakawa H, Cho JY, Libermann TA, Jin J, Harper JW, Munger K: Human papillomavirus type 16 E7 oncoprotein associated with the cullin 2 ubiquitin ligase complex, which contributes to degradation of the retinoblastoma tumor suppressor. J Virol 2007, 81:9737-9747.
• [9]Seo BJ, Mun MR, J RK, Kim CJ, Lee I, Chang YH, Park YH: Bile tolerant Lactobacillus reuteri isolated from pig feces inhibits enteric bacterial pathogens and porcine rotavirus. Vet Res Commun 2010, 34:323-333.
• [10]Ivec M, Botić T, Koren S, Jakobsen M, Weingartl H, Cencic A: Interactions of macrophages with probiotic bacteria lead to increased antiviral response against vesicular stomatitis virus. Antiviral Res 2007, 75:266-274.
• [11]Zhang W, Azevedo MS, Wen K, Gonzalez A, Saif LJ, Li G, Yousef AE, Yuan L: Probiotic Lactobacillus acidophilus enhances the immunogenicity of an oral rotavirus vaccine in gnotobiotic pigs. Vaccine 2008, 26:3655-3661.
• [12]Martín V, Maldonado A, Fernández L, Rodríguez JM, Connor RI: Inhibition of human immunodeficiency virus type 1 by lactic acid bacteria from human breastmilk. Breastfeed Med 2010, 5:153-158.
• [13]Yasui H, Kiyoshima J, Hori T: Reduction of influenza virus titer and protection against influenza virus infection in infant mice fed Lactobacillus casei Shirota. Clin Diagn Lab Immunol 2004, 11:675-679.
• [14]Yin YN, Yu QF, Fu N, Liu XW, Lu FG: Effects of four Bifidobacteria on obesity in high-fat diet induced rats. World J Gastroenterol 2010, 16:3394-3401.
• [15]Xiao JZ, Kondo S, Takahashi N, Miyaji K, Oshida K, Hiramatsu A, Iwatsuki K, Kokubo S, Hosono A: Effects of milk products fermented by Bifidobacterium longum on blood lipids in rats and healthy adult male volunteers. J Dairy Sci 2003, 86:2452-2461.
• [16]Le Leu RK, Hu Y, Brown IL, Woodman RJ, Young GP: Synbiotic intervention of Bifidobacterium lactis and resistant starch protects against colorectal cancer development in rats. Carcinogenesis 2010, 31:246-251.
• [17]Sekine K, Ohta J, Onishi M, Tatsuki T, Shimokawa Y, Toida T, Kawashima T, Hashimoto Y: Analysis of antitumor properties of effector cells stimulated with a cell wall preparation (WPG) of Bifidobacterium infantis. Biol Pharm Bull 1995, 18:148-153.
• [18]Scardovi V: Genus Bifidobacterium. In Bergey's Manual of Systemic Bacteriology. Edited by Krieg NR, Holt JG. Philadelphia, PA: Lippincott, Williams & Wilkins; 1986:1418-1434.
• [19]Ahn JB: Isolation and characterization of Bifidobacterium producing exopolysaccharide. Food Eng Prog 2005, 9:291-296.
• [20]Scheffner M, Romanczuk H, Münger K, Huibregtse JM, Mietz JA, Howley PM: Functions of human papillomavirus proteins. Curr Top Microbiol Immunol 1994, 186:83-99.
• [21]Hausen H: Papillomaviruses causing cancer: evasion from host-cell control in early events in carcinogenesis. J Natl Cancer Inst 2000, 92:690-698.
• [22]Luczak MW, Jagodzinski PP: Apicidin down-regulates human papillomavirus type 16 E6 and E7 transcripts and proteins in SiHa cervical cancer cells. Cancer Lett 2008, 272:53-60.
• [23]Kenneth JL, Thomas DS: Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 2001, 25:402-408.
• [24]González MA: Molecular biology of cervical cancer. Clin Transl Oncol 2007, 9:347-354.
• [25]Richardson H, Kelsall G, Tellier P, Voyer H, Abrahamowicz M, Ferenczy A, Coutlée F, Franco EL: The natural history of type-specific human papillomavirus infections in female university students. Cancer Epidemiol Biomarkers Prev 2003, 12:485-490.
• [26]Clifford GM, Rana RK, Franceschi S, Smith JS, Gough G, Pimenta JM: Human papillomavirus genotype distribution in low-grade cervical lesions: comparison by geographic region and with cervical cancer. Cancer Epidemiol Biomarkers Prev 2005, 14:1157-1164.
• [27]Vrese M, Rautenberg P, Laue C, Koopmans M, Herremans T, Schrezenmeir J: Probiotic bacteria stimulate virus-specific neutralizing antibodies following a booster polio vaccination. Eur J Nutr 2005, 44:406-413.
• [28]Cross ML, Ganner A, Teilab D, Fray LM: Patterns of cytokine induction by gram-positive and gram-negative probiotic bacteria. FEMS Immunol Med Microbiol 2004, 42:173-180.
• [29]Pohjavuori E, Viljanen M, Korpela R, Kuitunen M, Tiittanen M, Vaarala O, Savilahti E: Lactobacillus GG effect in increasing INF-gamma production in infants with cow's milk allergy. J Allergy Clin Immunol 2004, 114:131-136.
• [30]Dugas B, Mercenier A, Lenoir-Wijnkoop I, Arnaud C, Dugas N, Postaire E: Immunity and probiotics. Immunol Today 1999, 20:387-390.
• [31]Jagtar S, Shweta S: Classification, regulatory acts and applications of nutraceuticals for health. Int J Pharm Biol Sci 2012, 2:177-187.
• [32]Romond MB, Haddou Z, Mie lcareck C, Romond C: Bifidobacteria and human health: regulatory effect of indigenous bifidobacteria on Escherichia coli intestinal colonization. Anaerobe 1997, 3:131-136.
• [33]Wang KY, Li SN, Liu CS, Perng DS, Su YC, Wu DC, Jan CM, Lai CH, Wang TN, Wang WM: Effects of ingesting Lactobacillus- and Bifidobacterium containing yogurt in subjects with colonized Helicobacter pylori. Am J Clin Nutr 2004, 80:737-741.
• [34]Silva AM, Barbosa FH, Duarte R, Vieira LQ, Arantes RM, Nicoli JR: Effect of Bifidobacterium longum ingestion on experimental in salmonellosis in mice. J Appl Microbiol 2004, 97:29-37.
• [35]Woodman CB, Collins S, Winter H, Bailey A, Ellis J, Prior P, Yates M, Rollason TP, Young LS: Natural history of cervical human papillomavirus infection in young women: a longitudinal cohort study. Lancet 2001, 357:1831-1836.
• [36]Li GL, Jiang W, Xia Q, Chen SH, Ge XR, Gui SQ, Xu CJ: HPV E6 down-regulation and apoptosis induction of human cervical cancer cells by a novel lipid-soluble extract (PE) from Pinellia pedatisecta Schott in vitro. J Ethnopharmacol 2010, 132:56-64.