Login Register Cart 0
Generic placeholder image

Infectious Disorders - Drug Targets

Editor-in-Chief

ISSN (Print): 1871-5265
ISSN (Online): 2212-3989

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

Proliferative Effect of FadA Recombinant Protein from Fusobacterium nucleatum on SW480 Colorectal Cancer Cell Line

Author(s): Masoud Dadashi, Bahareh Hajikhani, Ebrahim Faghihloo, Parviz Owlia, Somayeh Yaslianifard, Mehdi Goudarzi, Mohammad Javad Nasiri and Fatemeh Fallah*

Volume 21, Issue 4, 2021

Published on: 20 July, 2020

Page: [623 - 628] Pages: 6

DOI: 10.2174/1871526520666200720113004

Price: $65

Abstract

Background and Aim: Colorectal Cancer (CRC) is one of the most frequent cancers diagnosed in both men and women worldwide. Fusobacterium nucleatum adhesin A (FadA) has an important potential factor in the development or progression of CRC. The aim of the present study was to evaluate the proliferative effect of recombinant FadA on SW480 colorectal cancer cell line.

Materials and Methods: The recombinant pET21(b)-fadA plasmid was synthesized and transformed into competent E.coli DH5α. In the next step, induction and expression of recombinant FadA were carried out in E. coli BL21 (DE3) competent cells. Expression and purification of protein were successfully done and it was analyzed and confirmed by SDS-PAGE and western blotting. The proliferative effect of purified FadA on SW480 CRC cell line was evaluated using MTT assay and cell counting methods.

Results: Visualization of the specific band isolated from the linear plasmid on the agarose gel confirmed the presence of the desired gene. After electrophoresis and Coomassie blue staining, the protein of interest with an approximate molecular weight of 13KDa was detected. The MTT assay, similar to cell counting methods, revealed that FadA dose and time-dependently promoted SW480 cell growth and proliferation in 24, 48 and 72 hours.

Conclusion: The results showed that FadA stimulates proliferation of SW480 colorectal cancer cell line with a dose and time-dependent manner.

Keywords: Fusobacterium nucleatum, colorectal cancer, adhesin A, SW480 Cell line, FadA, electrophoresis.

Graphical Abstract

[1]
Bhandari, A.; Woodhouse, M.; Gupta, S. Colorectal cancer is a leading cause of cancer incidence and mortality among adults younger than 50 years in the USA: a SEER-based analysis with comparison to other young-onset cancers. J. Investig. Med., 2017, 65(2), 311-315.
[http://dx.doi.org/10.1136/jim-2016-000229] [PMID: 27864324]
[2]
Connell, L.C.; Mota, J.M.; Braghiroli, M.I.; Hoff, P.M. The rising incidence of younger patients with colorectal cancer: questions about screening, biology, and treatment. Curr. Treat. Options Oncol., 2017, 18(4), 23.
[http://dx.doi.org/10.1007/s11864-017-0463-3] [PMID: 28391421]
[3]
Torre, L; Bray, F; Siegel, R; Ferlay, J; Lortet-Tieulent, J; Jemal, A. Global cancer statistics, 2012. CA A Cancer J Clin, 2015, 65, 87-108.
[4]
Sameer, A.S.S. Colorectal cancer: molecular mutations and polymorphisms. Front. Oncol., 2013, 3, 114.
[http://dx.doi.org/10.3389/fonc.2013.00114] [PMID: 23717813]
[5]
Ma, H.; Brosens, L.A.A.; Offerhaus, G.J.A.; Giardiello, F.M.; de Leng, W.W.J.; Montgomery, E.A. Pathology and genetics of hereditary colorectal cancer. Pathology, 2018, 50(1), 49-59.
[http://dx.doi.org/10.1016/j.pathol.2017.09.004] [PMID: 29169633]
[6]
Tóth, C.; Sükösd, F.; Valicsek, E.; Herpel, E.; Schirmacher, P.; Tiszlavicz, L. Loss of CDX2 gene expression is associated with DNA repair proteins and is a crucial member of the Wnt signaling pathway in liver metastasis of colorectal cancer. Oncol. Lett., 2018, 15(3), 3586-3593.
[PMID: 29467879]
[7]
Brocardo, M.; Henderson, B.R. APC shuttling to the membrane, nucleus and beyond. Trends Cell Biol., 2008, 18(12), 587-596.
[http://dx.doi.org/10.1016/j.tcb.2008.09.002] [PMID: 18848448]
[8]
Lee, D.W.; Han, S.W.; Kang, J.K.; Bae, J.M.; Kim, H-P.; Won, J-K.; Jeong, S.Y.; Park, K.J.; Kang, G.H.; Kim, T.Y. Association between Fusobacterium nucleatum, pathway mutation, and patient prognosis in colorectal cancer. Ann. Surg. Oncol., 2018, 25(11), 3389-3395.
[http://dx.doi.org/10.1245/s10434-018-6681-5] [PMID: 30062471]
[9]
Ma, C.T.; Luo, H.S.; Gao, F.; Tang, Q.C.; Chen, W. Fusobacterium nucleatum promotes the progression of colorectal cancer by interacting with E-cadherin. Oncol. Lett., 2018, 16(2), 2606-2612.
[http://dx.doi.org/10.3892/ol.2018.8947] [PMID: 30013655]
[10]
Ghosh, S.K.; Feng, Z.; Fujioka, H.; Lux, R.; McCormick, T.S.; Weinberg, A. Conceptual perspectives: bacterial antimicrobial peptide induction as a novel strategy for symbiosis with the human host. Front. Microbiol., 2018, 9, 302.
[http://dx.doi.org/10.3389/fmicb.2018.00302] [PMID: 29535688]
[11]
Hung, S.C.; Huang, P.R.; Almeida-da-Silva, C.L.C.; Atanasova, K.R.; Yilmaz, O.; Ojcius, D.M. NLRX1 modulates differentially NLRP3 inflammasome activation and NF-κB signaling during Fusobacterium nucleatum infection. Microbes Infect., 2018, 20(9-10), 615-625.
[http://dx.doi.org/10.1016/j.micinf.2017.09.014] [PMID: 29024797]
[12]
Dai, Z.; Zhang, J.; Wu, Q. The role of microbiota in the development of colorectal cancer. Int. J. Cancer, 2018.
[PMID: 30474116]
[13]
Walker, M.Y.; Pratap, S.; Southerland, J.H.; Farmer-Dixon, C.M.; Lakshmyya, K.; Gangula, P.R. Role of oral and gut microbiome in nitric oxide-mediated colon motility. Nitric Oxide, 2018, 73, 81-88.
[http://dx.doi.org/10.1016/j.niox.2017.06.003] [PMID: 28602746]
[14]
Liu, L.; Tabung, F.K.; Zhang, X.; Nowak, J.A.; Qian, Z.R.; Hamada, T.; Nevo, D.; Bullman, S.; Mima, K.; Kosumi, K.; da Silva, A.; Song, M.; Cao, Y.; Twombly, T.S.; Shi, Y.; Liu, H.; Gu, M.; Koh, H.; Li, W.; Du, C.; Chen, Y.; Li, C.; Li, W.; Mehta, R.S.; Wu, K.; Wang, M.; Kostic, A.D.; Giannakis, M.; Garrett, W.S.; Hutthenhower, C.; Chan, A.T.; Fuchs, C.S.; Nishihara, R.; Ogino, S.; Giovannucci, E.L. Diets That Promote Colon Inflammation Associate With Risk of Colorectal Carcinomas That Contain Fusobacterium nucleatum. Clin. Gastroenterol. Hepatol., 2018, 16(10), 1622-1631.e3.
[http://dx.doi.org/10.1016/j.cgh.2018.04.030] [PMID: 29702299]
[15]
Okita, Y.; Koi, M.; Koeppe, E.; Stoffel, E.M.; Galanko, J.; McCoy, N. 152-Fusobacterium Necleatum Infection Correlates with two Molecular Phenotypes of Colorectal Cancer (CRC) Associated with Inflammation: MSI-High and MSI-Low/EMAST. Gastroenterology, 2018, 154(6), S-41-S-2.
[http://dx.doi.org/10.1016/S0016-5085(18)30610-3]
[16]
Koi, M.; Okita, Y.; Carethers, J.M. Fusobacterium nucleatum infection in colorectal cancer: linking inflammation, dna mismatch repair and genetic and epigenetic alterations. J Anus Rectum Colon, 2018, 2(2), 37-46.
[http://dx.doi.org/10.23922/jarc.2017-055] [PMID: 30116794]
[17]
Yang, Y; Weng, W; Peng, J Fusobacterium nucleatum increases proliferation of colorectal cancer cells and tumor development in mice by activating toll-like receptor 4 signaling to nuclear factor− κb, and up-regulating expression of MicroRNA-21. Gastroenterology., 2017, 152(4), 851-866. e24..
[18]
Bashir, A.; Miskeen, A.Y.; Bhat, A.; Fazili, K.M.; Ganai, B.A. Fusobacterium nucleatum: an emerging bug in colorectal tumorigenesis. Eur. J. Cancer Prev., 2015, 24(5), 373-385.
[http://dx.doi.org/10.1097/CEJ.0000000000000116] [PMID: 25569450]
[19]
Fardini, Y.; Wang, X.; Témoin, S.; Nithianantham, S.; Lee, D.; Shoham, M.; Han, Y.W. Fusobacterium nucleatum adhesin FadA binds vascular endothelial cadherin and alters endothelial integrity. Mol. Microbiol., 2011, 82(6), 1468-1480.
[http://dx.doi.org/10.1111/j.1365-2958.2011.07905.x] [PMID: 22040113]
[20]
Rubinstein, M.R.; Wang, X.; Liu, W.; Hao, Y.; Cai, G.; Han, Y.W. Fusobacterium nucleatum promotes colorectal carcinogenesis by modulating E-cadherin/β-catenin signaling via its FadA adhesin. Cell Host Microbe, 2013, 14(2), 195-206.
[http://dx.doi.org/10.1016/j.chom.2013.07.012] [PMID: 23954158]
[21]
Keku, T.O.; McCoy, A.N.; Azcarate-Peril, A.M. Fusobacterium spp. and colorectal cancer: cause or consequence? Trends Microbiol., 2013, 21(10), 506-508.
[http://dx.doi.org/10.1016/j.tim.2013.08.004] [PMID: 24029382]
[22]
Shang, F-M.; Liu, H-L. Fusobacterium nucleatum and colorectal cancer: A review. World J. Gastrointest. Oncol., 2018, 10(3), 71-81.
[http://dx.doi.org/10.4251/wjgo.v10.i3.71] [PMID: 29564037]
[23]
Kumar, A.; Thotakura, P.L.; Tiwary, B.K.; Krishna, R. Target identification in Fusobacterium nucleatum by subtractive genomics approach and enrichment analysis of host-pathogen protein-protein interactions. BMC Microbiol., 2016, 16(1), 84.
[http://dx.doi.org/10.1186/s12866-016-0700-0] [PMID: 27176600]
[24]
Zhang, S.; Cai, S.; Ma, Y. Association between Fusobacterium nucleatum and colorectal cancer: Progress and future directions. J. Cancer, 2018, 9(9), 1652-1659.
[http://dx.doi.org/10.7150/jca.24048] [PMID: 29760804]
[25]
Xu, M.; Yamada, M.; Li, M.; Liu, H.; Chen, S.G.; Han, Y.W. FadA from Fusobacterium nucleatum utilizes both secreted and nonsecreted forms for functional oligomerization for attachment and invasion of host cells. J. Biol. Chem., 2007, 282(34), 25000-25009.
[http://dx.doi.org/10.1074/jbc.M611567200] [PMID: 17588948]
[26]
Riki, M.; Farokhi, F.; Tukmechi, A. The best time of cytotoxicity for extracted cell wall from Lactobacillus casei and paracasei in K562 cell line. Tehran Univ. Med. J., 2013, 70(11), 691-699.
[27]
Haggar, F.A.; Boushey, R.P. Colorectal cancer epidemiology: incidence, mortality, survival, and risk factors. Clin. Colon Rectal Surg., 2009, 22(4), 191-197.
[http://dx.doi.org/10.1055/s-0029-1242458] [PMID: 21037809]
[28]
Ito, M.; Kanno, S.; Nosho, K.; Sukawa, Y.; Mitsuhashi, K.; Kurihara, H.; Igarashi, H.; Takahashi, T.; Tachibana, M.; Takahashi, H.; Yoshii, S.; Takenouchi, T.; Hasegawa, T.; Okita, K.; Hirata, K.; Maruyama, R.; Suzuki, H.; Imai, K.; Yamamoto, H.; Shinomura, Y. Association of Fusobacterium nucleatum with clinical and molecular features in colorectal serrated pathway. Int. J. Cancer, 2015, 137(6), 1258-1268.
[http://dx.doi.org/10.1002/ijc.29488] [PMID: 25703934]
[29]
McCoy, A.N.; Araújo-Pérez, F.; Azcárate-Peril, A.; Yeh, J.J.; Sandler, R.S.; Keku, T.O. Fusobacterium is associated with colorectal adenomas. PLoS One, 2013, 8(1), e53653.
[http://dx.doi.org/10.1371/journal.pone.0053653] [PMID: 23335968]
[30]
Leader, B.; Baca, Q.J.; Golan, D.E. Protein therapeutics: a summary and pharmacological classification. Nat. Rev. Drug Discov., 2008, 7(1), 21-39.
[http://dx.doi.org/10.1038/nrd2399] [PMID: 18097458]
[31]
Kostic, A.D.; Chun, E.; Robertson, L.; Glickman, J.N.; Gallini, C.A.; Michaud, M.; Clancy, T.E.; Chung, D.C.; Lochhead, P.; Hold, G.L.; El-Omar, E.M.; Brenner, D.; Fuchs, C.S.; Meyerson, M.; Garrett, W.S. Fusobacterium nucleatum potentiates intestinal tumorigenesis and modulates the tumor-immune microenvironment. Cell Host Microbe, 2013, 14(2), 207-215.
[http://dx.doi.org/10.1016/j.chom.2013.07.007] [PMID: 23954159]
[32]
Mima, K.; Nishihara, R.; Qian, Z.R.; Cao, Y.; Sukawa, Y.; Nowak, J.A.; Yang, J.; Dou, R.; Masugi, Y.; Song, M.; Kostic, A.D.; Giannakis, M.; Bullman, S.; Milner, D.A.; Baba, H.; Giovannucci, E.L.; Garraway, L.A.; Freeman, G.J.; Dranoff, G.; Garrett, W.S.; Huttenhower, C.; Meyerson, M.; Meyerhardt, J.A.; Chan, A.T.; Fuchs, C.S.; Ogino, S. Fusobacterium nucleatum in colorectal carcinoma tissue and patient prognosis. Gut, 2016, 65(12), 1973-1980.
[http://dx.doi.org/10.1136/gutjnl-2015-310101] [PMID: 26311717]

Rights & Permissions Print Cite
Article Metrics
39
1
© 2024 Bentham Science Publishers | Privacy Policy