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Live and heat-killed Lactobacillus rhamnosus GG upregulate gene expression of pro-inflammatory cytokines in 5-fluorouracil-pretreated Caco-2 cells

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Abstract

Purpose

This study investigates whether post-chemotherapeutic use of live and heat-killed Lactobacillus rhamnosus GG can modulate the expression of three pro-inflammatory cytokines in 5-fluorouracil (5-FU)-induced intestinal mucositis in vitro.

Methods

Live L. rhamnosus GG and heat-killed L. rhamnosus GG were observed using scanning electron microscopy. To establish the duration required for optimal expression of tumor necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1), and interleukin-12 (IL-12), 5 μM of 5-FU was selected to treat 10-day-old Caco-2 cells for 4, 6, 8, and 24 h. Caco-2 cells were treated with 5-FU (5 μM) for 4 h, followed by the administration of live L. rhamnosus GG (multiplicity of infection = 25), and heat-killed L. rhamnosus GG for 2 and 4 h. Finally, total cellular RNA was isolated to quantify mRNA expression of TNF-α, MCP-1, and IL-12 using real-time PCR.

Results

The results demonstrated that heat-killed L. rhamnosus GG remained structurally intact with elongation. A biphasic upregulated expression of TNF-α, MCP-1, and IL-12 was observed in 5-FU-treated Caco-2 cells at 4 and 24 h. Compared to non-L. rhamnosus GG controls in 5-FU-pretreated Caco-2 cells, a 2-h treatment of heat-killed L. rhamnosus GG significantly upregulated the MCP-1 expression (p < 0.05), and both live and heat-killed L. rhamnosus GG treatments lasting 4 h upregulated the TNF-α and MCP-1 expression (p < 0.05). Only live L. rhamnosus GG upregulated the IL-12 expression (p < 0.05).

Conclusions

Post-chemotherapeutic use of live or heat-killed L. rhamnosus GG can upregulate the gene expression of 5-FU-induced pro-inflammatory cytokines in Caco-2 cells. Human intestinal epithelium may be vulnerable to the post-chemotherapeutic use of L. rhamnosus GG in 5-FU-induced mucositis that requires further in vivo studies for clarification.

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Acknowledgments

This study was funded by the Taipei Medical University (TMU101-AE1-B64), the National Health Research Institutes (ME-101-PP-12), and the National Taipei University of Technology (NTUT-MMH-101-07). We thank Professor Liang-Shun Wang, Dr. Wei-Ming Chi, and Dr. Chi-Tai Yeh for their assistance in the laboratory.

Conflict of interest

I, Shiuh-Bin Fang, state that there is no conflict of interest associated with this manuscript. I declare that I have no financial relationship with the organizations that funded the research. The authorship of this manuscript is as described in the attached Authorship/Disclosure form, with myself as the first author. I have full control of all primary data related to this manuscript and agree to allow the journal to review the data if necessary.

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Authors and Affiliations

  1. Division of Pediatric Gastroenterology and Hepatology, Department of Pediatrics, Shuang Ho Hospital, Taipei Medical University, 291, Jhong Jheng Road, Jhong Ho District, Taipei, 23561, Taiwan

    Shiuh-Bin Fang

  2. Department of Pediatrics, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan

    Shiuh-Bin Fang

  3. Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology, 1, Section 3, Chung Hsiao East Road, Taipei 106, Taiwan

    Hsin-Yu Shih, Chih-Hung Huang, Li-Ting Li, Chia-Chun Chen & Hsu-Wei Fang

  4. Division of Medical Engineering Research, National Health Research Institutes, Miaoli, Taiwan

    Hsu-Wei Fang

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  1. Shiuh-Bin Fang
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Correspondence to Shiuh-Bin Fang or Hsu-Wei Fang.

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Fang, SB., Shih, HY., Huang, CH. et al. Live and heat-killed Lactobacillus rhamnosus GG upregulate gene expression of pro-inflammatory cytokines in 5-fluorouracil-pretreated Caco-2 cells. Support Care Cancer 22, 1647–1654 (2014). https://doi.org/10.1007/s00520-014-2137-z

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  • DOI: https://doi.org/10.1007/s00520-014-2137-z

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