Berberin inflamasyon baskılanması ve apoptoz indüksiyonu ile Caco-2 kolorektal adenokarsinom hücrelerinde 5-florourasil'in terapötik etkisini arttırır

Year 2023, Volume: 48 Issue: 4, 1238 - 1247, 29.12.2023

Seyma Oncu , Merve Becit-kizilkaya , Serkan Şen , Fatma Özlem Kargın Solmaz , Sefa Çelik

https://doi.org/10.17826/cumj.1344952

Cited By: 1

Abstract

Amaç: Bu çalışma, berberin (BBR) ile kemoterapinin omurgasını oluşturan 5-fluorourasil (5FU)’in Caco-2 kolorektal adenokarsinoma hücreleri üzerinde sinerjik etki gösterip göstermediğini ve olası mekanizmalarını araştırmayı amaçlamaktadır.
Gereç ve Yöntem: Hücre canlılığı MTT yöntemiyle ve kombinasyon indeksi (CI) Chou-Talalay yöntemiyle belirlendi. Apoptoz ve inflamasyonla ilişkili proteinlerin (Bcl-2, p53, IL-6, TNF-α, MMP-9) konsantrasyonları ELISA ile ölçüldü.
Bulgular: BBR ve 5FU’nun IC50 değerleri 24 saat için 280 µM ve 20 mM olarak bulundu. Kombinasyon tedavi, her ikisinin de IC50 konsantrasyonlarında daha belirgin olmak üzere, sinerjik sitotoksik etki gösterdi (CI= 0.143). 5FU+BBR, Bcl-2’yi azaltarak ve p53’ü artırarak sinerjistik apoptotik etki gösterdi (5FU ile karşılaştırıldığında, Bcl-2 için 0.712 kat azalış ve p53 için 2.650 kat artış). Ayrıca, 5FU ile artan IL-6 ve TNF-α düzeyleri, 5FU+BBR tedavisi ile belirgin şekilde baskılandı (0.733 kat ve 0.485 kat). Ek olarak, 5FU ile tedavi edilen hücrelerde kontrol grubuna göre MMP-9 düzeylerinde anlamlı bir fark yokken, kombine tedavi MMP-9 düzeylerini belirgin şekilde (0.601 kat) azalttı.
Sonuç: Bulgular, BBR’nin 5FU ile birlikte kolorektal kanseri kemoterapisi için apoptoz indüksiyonu, inflamasyon baskılaması ve metastaz inhibisyonuyla muazzam potansiyelini ortaya koymaktadır.

Keywords

Berberin (BBR), 5-fluorourasil (5FU), sinerjizm, Caco-2, apoptosis, MMP-9

Berberine enhances the therapeutic effect of 5-fluorouracil in Caco-2 colorectal adenocarcinoma cells by alleviating inflammation and inducing apoptosis

Abstract

Purpose: This study aims to investigate whether berberine (BBR) and 5-fluorouracil (5FU), which forms the backbone of chemotherapy, have a synergistic effect on colorectal adenocarcinoma cells (Caco-2) and what mechanisms might be behind this.
Materials and Methods: Cell viability was determined by MTT and the combination index (CI) by Chou-Talalay method. Apoptosis and inflammation-related proteins (Bcl-2, p53, IL-6, TNF-α, MMP-9) were measured by ELISA.
Results: The IC50 values of BBR and 5FU were found to be 280 µM and 20 mM for 24 h. The combination treatment showed synergistic cytotoxicity, both of which were more pronounced at IC50 (CI=0.143). 5FU+BBR showed a synergistic apoptotic effect by decreasing Bcl-2 and increasing p53 (0.712-fold decrease in Bcl-2 and 2.650-fold increase in p53 compared to 5FU). The increase in IL-6 and TNF-α by 5FU was significantly suppressed by 5FU+BBR (0.733-fold and 0.485-fold). Although there was no significant difference in MMP-9 in 5FU compared to control, 5FU+BBR significantly decreased MMP-9 (0.601-fold).
Conclusion: The results show the enormous potential of BBR in combination with 5FU for the chemotherapy of colorectal carcinoma through apoptosis induction, inflammation inhibition and metastasis inhibition.

Keywords

berberine (BBR), 5-fluorouracil (5FU), synergism, Caco-2, apoptosis, MMP-9

References

• Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021;71:209-49.
• Xi Y, Xu P. Global colorectal cancer burden in 2020 and projections to 2040. Transl Oncol. 2021;14:101174.
• Sałaga M, Zatorski H, Sobczak M, Chen C, Fichna J. Chinese herbal medicines in the treatment of IBD and colorectal cancer: a review. Curr Treat Options Oncol. 2014;15:405-20.
• Zhang Y, Chen Z, Li J. The current status of treatment for colorectal cancer in China: A systematic review. Medicine (Baltimore). 2017;96:e8242.
• Yau TO. Precision treatment in colorectal cancer: Now and the future. JGH Open. 2019;3:361-69.
• Entezar-Almahdi E, Mohammadi-Samani S, Tayebi L, Farjadian F. Recent advances in designing 5-Fluorouracil delivery systems: A stepping stone in the safe treatment of colorectal cancer. Int J Nanomedicine. 2020;15:5445-58.
• Lee CS, Ryan EJ, Doherty GA. Gastro-intestinal toxicity of chemotherapeutics in colorectal cancer: the role of inflammation. World J Gastroenterol. 2014;20:3751-61.
• Ciombor KK, Bekaii-Saab T. A comprehensive review of sequencing and combination strategies of targeted agents in metastatic colorectal cancer. Oncologist. 2018;23:25-34.
• Kong MY, Li LY, Lou YM, Chi HY, Wu JJ. Chinese herbal medicines for prevention and treatment of colorectal cancer: From molecular mechanisms to potential clinical applications. J Integr Med. 2020;18:369-84.
• Jiang X, Jiang Z, Jiang M, Sun Y. Berberine as a potential agent for the treatment of colorectal cancer. Front Med (Lausanne). 2022;9:886996.
• Hallajzadeh J, Maleki Dana P, Mobini M, Asemi Z, Mansournia MA, Sharifi M et al. Targeting of oncogenic signaling pathways by berberine for treatment of colorectal cancer. Med Oncol. 2020;37:49.
• Chen YX, Gao QY, Zou TH, Wang BM, Liu SD, Sheng JQ et al. Berberine versus placebo for the prevention of recurrence of colorectal adenoma: a multicentre, double-blinded, randomised controlled study. Lancet Gastroenterol Hepatol. 2020;5:267-75.
• Li P, Hao Z, Liu H, Zhu B, Dang L, Ma C et al. Quantitative proteomics analysis of berberine-treated colon cancer cells reveals potential therapy targets. Biology (Basel). 2021;10:250.
• Kwon S, Chan AT. Extracting the benefits of berberine for colorectal cancer. Lancet Gastroenterol Hepatol. 2020;5:231-33.
• Tong M, Liu H, Hao J, Fan D. Comparative pharmacoproteomics reveals potential targets for berberine, a promising therapy for colorectal cancer. Biochem Biophys Res Commun. 2020:S0006-291X(20)30320-X.
• Rauf A, Abu-Izneid T, Khalil AA, Imran M, Shah ZA, Emran TB et al. Berberine as a potential anticancer agent: a comprehensive review. Molecules. 2021;26:7368.
• Dai W, Mu L, Cui Y, Li Y, Chen P, Xie H et al. Berberine promotes apoptosis of colorectal cancer via regulation of the long non-coding RNA (lncRNA) Cancer Susceptibility Candidate 2 (CASC2)/AU-Binding Factor 1 (AUF1)/B-Cell CLL/lymphoma 2 (Bcl-2) Axis. Med Sci Monit. 2019;25:730-38.
• Gong C, Hu X, Xu Y, Yang J, Zong L, Wang C et al. Berberine inhibits proliferation and migration of colorectal cancer cells by downregulation of GRP78. Anticancer Drugs. 2020;31:141-49.
• Lü Y, Han B, Yu H, Cui Z, Li Z, Wang J. Berberine regulates the microRNA-21-ITGΒ4-PDCD4 axis and inhibits colon cancer viability. Oncol Lett. 2018;15:5971-76.
• Wang L, Cao H, Lu N, Liu L, Wang B, Hu T et al. Berberine inhibits proliferation and down-regulates epidermal growth factor receptor through activation of Cbl in colon tumor cells. PLoS One. 2013;8:e56666.
• Samad MA, Saiman MZ, Abdul Majid N, Karsani SA, Yaacob JS. Berberine inhibits telomerase activity and induces cell cycle arrest and telomere erosion in colorectal cancer cell line, HCT 116. Molecules. 2021;26:376.
• Liu X, Ji Q, Ye N, Sui H, Zhou L, Zhu H et al. Berberine inhibits invasion and metastasis of colorectal cancer cells via COX-2/PGE2 mediated JAK2/STAT3 signaling pathway. PLoS One. 2015;10:e0123478.
• Vivarelli S, Salemi R, Candido S, Falzone L, Santagati M, Stefani S et al. Gut microbiota and cancer: from pathogenesis to therapy. Cancers (Basel). 2019;11:38.
• Habtemariam S. Berberine pharmacology and the gut microbiota: A hidden therapeutic link. Pharmacol Res. 2020;155:104722.
• Yu M, Tong X, Qi B, Qu H, Dong S, Yu B et al. Berberine enhances chemosensitivity to irinotecan in colon cancer via inhibition of NF‑κB. Mol Med Rep. 2014;9:249-54.
• Devarajan N, Jayaraman S, Mahendra J, Venkatratnam P, Rajagopal P, Palaniappan H et al. Berberine-A potent chemosensitizer and chemoprotector to conventional cancer therapies. Phytother Res. 2021;35:3059-77.
• Bao J, Huang B, Zou L, Chen S, Zhang C, Zhang Y et al. Hormetic effect of berberine attenuates the anticancer activity of chemotherapeutic agents. PLoS One. 2015;10:e0139298.
• Becit-Kızılkaya M, Öncü Ş, Şen S, Çelik S (2023) Berberine synergizes with cisplatin via ınducing apoptosis on A549 non-small cell lung cancer cells. Eur J Ther. 2023;29:480-88.
• Pandey A, Vishnoi K, Mahata S, Tripathi SC, Misra SP, Misra V et al. Berberine and curcumin target survivin and STAT3 in gastric cancer cells and synergize actions of standard chemotherapeutic 5-Fluorouracil. Nutr Cancer. 2015;67:1293-304.
• Palmieri A, Iapichino A, Cura F, Scapoli L, Carinci F, Mandrone M et al. Pre-treatment with berberine enhances effect of 5-fluorouracil and cisplatin in HEP2 laryngeal cancer cell line. J Biol Regul Homeost Agents. 2018;32:167-177.
• He C, Rong R, Liu J, Wan J, Zhou K, Kang JX. Effects of Coptis extract combined with chemotherapeutic agents on ROS production, multidrug resistance, and cell growth in A549 human lung cancer cells. Chin Med. 2012;7:11.
• Cai Y, Xia Q, Luo R, Huang P, Sun Y, Shi Y et al. Berberine inhibits the growth of human colorectal adenocarcinoma in vitro and in vivo. J Nat Med. 2014;68:53-62.
• Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65:55-63.
• Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 2010;70:440-6.
• Bishayee A, Sethi G. Bioactive natural products in cancer prevention and therapy: Progress and promise. Semin Cancer Biol. 2016;40-41:1-3.
• Negrette-Guzmán M. Combinations of the antioxidants sulforaphane or curcumin and the conventional antineoplastics cisplatin or doxorubicin as prospects for anticancer chemotherapy. Eur J Pharmacol. 2019;859:172513.
• Liu Y, Hua W, Li Y, Xian X, Zhao Z, Liu C et al. Berberine suppresses colon cancer cell proliferation by inhibiting the SCAP/SREBP-1 signaling pathway-mediated lipogenesis. Biochem Pharmacol. 2020;174:113776.
• Ramesh P, Medema JP. BCL-2 family deregulation in colorectal cancer: potential for BH3 mimetics in therapy. Apoptosis. 2020;25:305-20.
• Huang D, Sun W, Zhou Y, Li P, Chen F, Chen H et al. Mutations of key driver genes in colorectal cancer progression and metastasis. Cancer Metastasis Rev. 2018;37:173-87.
• Wang N, Feng Y, Zhu M, Tsang CM, Man K, Tong Y et al. Berberine induces autophagic cell death and mitochondrial apoptosis in liver cancer cells: the cellular mechanism. J Cell Biochem. 2010;111:1426-36.
• Okubo S, Uto T, Goto A, Tanaka H, Nishioku T, Yamada K et al. Berberine Induces Apoptotic Cell Death via Activation of Caspase-3 and -8 in HL-60 Human Leukemia Cells: Nuclear Localization and Structure-Activity Relationships. Am J Chin Med. 2017;45:1497-1511.
• Li DX, Zhang J, Zhang Y, Zhao PW, Yang LM. Inhibitory effect of berberine on human skin squamous cell carcinoma A431 cells. Genet Mol Res. 2015;14:10553-68.
• Shi XZ, Zhao S, Wang Y, Wang MY, Su SW, Wu YZ et al. Antitumor activity of berberine by activating autophagy and apoptosis in CAL-62 and BHT-101 anaplastic thyroid carcinoma cell lines. Drug Des Devel Ther. 2023;17:1889-1906.
• Todoric J, Antonucci L, Karin M. Targeting inflammation in cancer prevention and therapy. Cancer Prev Res (Phila). 2016;9:895-905.
• Deng J, Zhao L, Yuan X, Li Y, Shi J, Zhang H et al. Pre-Administration of berberine exerts chemopreventive effects in AOM/DSS-induced colitis-associated carcinogenesis mice via modulating inflammation and intestinal microbiota. Nutrients. 2022;14:726.
• Chen J, Pitmon E, Wang K. Microbiome, inflammation and colorectal cancer. Semin Immunol. 2017;32:43–53.
• Francescone R, Hou V, Grivennikov SI. Cytokines, IBD, and colitis-associated cancer. Inflamm Bowel Dis. 2015;21:409-18.
• Lou Y, Song M, Han M, Zhong J, Tian X, Ren Y et al. Tumor Necrosis Factor-α-Induced Protein 8-Like 2 fosters Tumor-Associated Microbiota to promote the development of colorectal cancer. Cancer Immunol Res. 2022;10:354-67.
• Chen H, Zhang F, Li R, Liu Y, Wang X, Zhang X et al. Berberine regulates fecal metabolites to ameliorate 5-fluorouracil induced intestinal mucositis through modulating gut microbiota. Biomed Pharmacother. 2020;124:109829.
• Chen H, Ye C, Cai B, Zhang F, Wang X, Zhang J et al. Berberine inhibits intestinal carcinogenesis by suppressing intestinal pro-inflammatory genes and oncogenic factors through modulating gut microbiota. BMC Cancer. 2022;22:566.
• Jonsson A, Hjalmarsson C, Falk P, Ivarsson ML. Stability of matrix metalloproteinase-9 as biological marker in colorectal cancer. Med Oncol. 2018;35:50.
• Chu SC, Yu CC, Hsu LS, Chen KS, Su MY, Chen PN. Berberine reverses epithelial-to-mesenchymal transition and inhibits metastasis and tumor-induced angiogenesis in human cervical cancer cells. Mol Pharmacol. 2014;86:609-23.
• Liu JF, Lai KC, Peng SF, Maraming P, Huang YP, Huang AC et al. Berberine inhibits human melanoma A375.S2 cell öigration and invasion via affecting the FAK, uPA, and NF-κB signaling pathways and inhibits PLX4032 resistant A375.S2 cell öigration in vitro. Molecules. 2018;23:2019.
• Chen W, Miao YQ, Fan DJ, Yang SS, Lin X, Meng LK et al. Bioavailability study of berberine and the enhancing effects of TPGS on intestinal absorption in rats. AAPS PharmSciTech. 2011;12:705–11.
• Majidzadeh H, Araj-Khodaei M, Ghaffari M, Torbati M, Ezzati Nazhad Dolatabadi J, Hamblin MR. Nano-based delivery systems for berberine: A modern anti-cancer herbal medicine. Colloids Surf B Biointerfaces. 2020;194:111188