Commentary Open Access
Volume 4 | Issue 2 | DOI: https://doi.org/10.33696/cancerimmunol.4.064

Can Butein be a Future Candidate for the Treatment of Advance Metastatic Thyroid Cancer?

  • 1Radiation Medicine Centre, Bhabha Atomic Research Centre, c/o TMH Annexe, Parel, Mumbai, India
  • 2Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai, India
+ Affiliations - Affiliations

*Corresponding Author

Savita Kulkarni, savita.kulkarni1@gmail.com; savitapk@barc.gov.in

Received Date: April 19, 2022

Accepted Date: August 04, 2022


The incidence and prevalence of papillary thyroid cancer (PTC) are increasing worldwide and it is the 5th most common endocrine cancer in females [1]. In addition to this, the frequency of resistance toward radio-iodine therapy is also increasing in PTC patients (Advanced metastatic thyroid cancer). External beam radiation therapy (EBRT) and chemotherapy are used for the treatment of such patients. EBRT and Chemotherapy are associated with serious side effects and toxicity. US-FDA has also approved two drugs (Sorafenib and Lenvatinib) for the treatment of advanced thyroid cancer patients. However, the efficacy of both drugs is limited in terms of overall survival and disease-free survival and associated with severe toxicities [2]. Hence, the treatment of patients with advanced metastatic thyroid cancer represents a major challenge for clinicians and oncologists. In such patients, tumor cells show invasion in local neck regions, lungs, and bones [3]. Metastasis is the most dangerous aspect of cancer and is responsible for 90% of deaths of cancer. Epithelial -mesenchymal transition (EMT) and cancer stem cells (CSC) are the driving forces of metastasis and therapeutic resistance [4]. Hence, this axis of EMT and CSCs is a major target from the new therapy point of view.

One approach for novel cancer therapeutics is to look for plant-based phytochemicals, especially flavonoids. Extensive studies were performed to explore the cytotoxic and antimetastatic effects of flavonoids. Flavonoids may exert an anti-metastatic effect by reversing EMT or modulation of EMT governing signaling pathways. Several reviews have shown that different subgroups of flavonoids reduce the expression of mesenchymal markers like vimentin, N-cadherin, matrix metalloproteinase, increase E-cadherin expression, and target CSCs [5,6]. Chalcones, α, and β unsaturated ketones, are one of the most important bioactive flavonoids with therapeutic potential linked to a variety of biological effects studied in various in-vitro and in-vivo investigations [7]. From ancient times plant-derived chalcones especially butein is used for the treatment of several diseases. Butein is extracted from Buteamonosperma, Rhus-verniciflua, and Semecarpus- anacardium. It has been reported to have antimicrobial, antioxidant, antiviral, anti-inflammatory, cytotoxic, anti-malarial, antidiabetic, and vasodilatory effects and is used as herb in South East Asia region. Further, reports have also shown the anticancerous effect of butein in various cancer models [8]. It was also demonstrated that in thyroid cancer, butein induces an intrinsic pathway of apoptosis, proteolysis of vimentin, and inhibits cancer stem cells in the human papillary thyroid cancer cell line. Additionally, in the paper by Tripathi & Kulkarni [7], the effect of butein at low and high doses was also demonstrated. At low doses, butein induces inhibition of cell migration and cancer stem cells through suppression of enhanced glycolysis and vimentin phosphorylation. While butein induces caspase activation and downregulates anti-apoptotic protein in the NPA cell line for the execution of the intrinsic pathway of apoptosis and vimentin proteolysis at high concentration [9].

The study has further raised important questions which are discussed sequentially: to be addressed. 1) mechanism behind the low and high doses response of butein, 2) long term exposure of butein and its relation with EMT and CSCs mediated cancer resistance, 3) Butein and cancer cell metabolism with reference to EMT and CSCs, 4) Safety of butein to normal cells and 5) Probability of utilization of butein as an anti-cancer therapeutic agent for the thyroid cancer or to increase the efficacy of present therapeutic regime.

The differential dose-response is a dynamic process or mechanism and depends upon the concentration and time duration. For example- curcumin exerts anti-oxidant properties like reactive oxygen species scavenging, activation of antioxidant enzymes, anti-apoptotic proteins, and heat shock proteins at low dose while induces DNA damage, apoptosis and pro-oxidant property at high dose. One of the mechanisms of the differential response of curcumin based on metabolic profiling of MCF-7 and MDA-MB-231 is the biphasic levels of glutathione in cancer cells (high level of glutathione at low dose and low level of glutathione at high dose) [10]. Butein also increases total GSH levels at 5, 10 and 25 μM in hepatocytes and protects them from tert-butylhydroperoxide (tBHP) mediated oxidative stress through ERK/Nrf2 pathway [11]. Similarly, butein increases heme oxygenase activity through Nrf2/ARE pathway to prevent H2O2 induced cell death in dental pulpal cells at 2.5, 5, 10 and 20 μM concentrations [12]. Butein may influence the redox status of cells in a differential way depending upon the dose and time duration. Hence, it is important to explore and decipher the effect of different doses of butein in suitable thyroid cancer models for the assessment of redox status, cell-protective signaling pathways (ARE/ Nrf2 pathways), and cell-protective proteins to further understand the mechanism of action at low and high doses of butein.

Long-term exposure to chemotherapeutic agents especially at low doses is associated with the development of resistant clones in cancer cells which finally leads to decreased response toward standard therapies [13]. Studies have further shown that exposure to chemotherapy leads to EMT activation and an increase in CSCs [14,15]. EMT and CSCs axis is the major reason behind the metastasis, recurrence, and therapeutic resistance [16]. EMT plays an important role in the acquisition of mesenchymal characteristics like vimentin and CSCs emergence. In PTC, vimentin is linked with invasion, migration, and poor prognosis [17]. In directional migration, vimentin interacts with microtubules and actin filaments and co-ordinate their dynamics. Microtubules contribute to the polarity in the migratory cells and Microtubules have a shorter life in comparison to vimentin. During cell migration, vimentin disassembles from the periphery and undergoes retrograde transport to incorporate into the mature filaments. This retrograde flow of vimentin helps in restricting actin retrograde flow and prevents nucleus collapse. Disassembly of vimentin from the periphery of the cell act as template for the growing microtubules and maintaining the cell polarity. Thus, vimentin acts as a template and guide the growing microtubules for the maintenance of cell polarity [18,19]. In a cancer thyroid cell, Merca cell line (A murine cell line developed from-Braf V600E mice), ectopic expression of Snail (EMT transcription factor) induces in vimentin and CSCs upregulation. This induction of EMT and stemness was significantly inhibited by Celastrol a natural inhibitor of neoplastic cells [20].

Vimentin’s role in cancer depends upon the structure and post-translational modification (PTM) status of the vimentin. Phosphorylation of vimentin is the most important PTM. Eriksson et al. characterized the 32P labeled phosphovimentin filament and showed the critical serine and threonine residues are important for phosphorylation. They further reported in BHK-1 fibroblast cells that protein phosphatase inhibition with calyculin A resulted in fast vimentin phosphorylation and phosphorylation induced disassembly into the soluble tetrameric vimentin oligomer [21]. Similarly, in serum-starved fibroblast, vimentin filaments were shown to get assembled in the periphery region. Activation of Rac1 in serum-starved fibroblast phosphorylates vimentin at the ser-38 residue. Phosphorylation induces depolymerization and retraction of vimentin from the cell surface where lamellipodia formation take place [22]. Various protein kinases like PKA, PKB, PKC, p21 activated kinase, Aurora B, RhoA-binding kinase α, and CaMkII; phosphorylate vimentin protein at various serine residues and influence the assembly of the vimentin filaments [21,23]. Ivaska et al. had shown that PKC mediated vimentin phosphorylation is important for the integrin trafficking and cell migration towards the matrix [24]. Hence, vimentin is suitable target for the development of an effective anti-metastatic agent.

Our study demonstrated that Butein induces inhibition of vimentin phosphorylation and further induces caspase -mediated proteolysis in the NPA cell line [7]. Zhu et al., had shown that protein kinase B (Akt) directly interacts with vimentin and induces serine 39 phosphorylation. Inhibition of vimentin phosphorylation or substitution of serine with alanine not only suppresses cancer cell migration but also induces caspase-mediated proteolysis [25]. Further, vimentin is important for cell division and stemness. A small molecule like FiVe1 binds with vimentin and targets stemness in mesenchymal cancer cells [26]. Thus, the development of small molecule inhibitors which directly target the activity of vimentin is a good approach for the development of antimetastatic therapeutic agents. Hence, it is important to study the interaction between vimentin and butein to decipher the mechanism of the anti-metastatic effect. Butein may directly interact with vimentin domains to inhibit its phosphorylation or butein may inhibit activities of several protein kinases for the inhibition of vimentin phosphorylation. However, in silico and in vitro studies are required to establish the interaction between vimentin and butein in more depth to decipher the mechanism of butein at low and high concentrations in more kinetic and structural approach.

An altered metabolism plays an important role in cancer cell proliferation, migration, invasion, EMT, drug resistance and CSC maintenance. Cancer cells displayed altered glucose metabolism, glutamine metabolism and also changes the regulation of key enzymes for the metastasis, drug resistance and CSCs [27,28]. Several oncogenes play an important role in cancer metabolism. In thyroid cancer, epidermal growth factor receptor (EGFR) alternation is linked with dedifferentiation, EMT and metastasis [29]. EGFR signaling also influence various metabolic functions like glucose utilization to fatty acids and nucleotide synthesis [30]. Hence, it is important to inspect effect of butein on cancer cell metabolism and genetic alternations. In hepatocellular carcinoma, butein inhibits EGFR and hexokinase-2 interaction to inhibit the glycolysis and proliferation [31]. Similarly, butein inhibits the glucose induced cell proliferation, induces ROS, DNA damage and p38 activation to induce anti-cancerous effect on non-small lung cancer cells [32]. Our study on NPA cell line also showed that butein inhibits glycolysis at low concentration [9]. Hence, it is important to investigate effect of butein on metabolic changes, EGFR and its link with genetic changes in thyroid cancer to reveal the mechanism through metabolomics approach.

Activation of EMT leads to the process of dedifferentiation in cancer cells. Dedifferentiation of thyroid cancer leads to loss of sodium iodide transporter and decreased radioiodine uptake [33,34]. Hence, long-term exposure to butein should be explored in suitable thyroid cancer models especially in radioiodine concentrating and radioiodine non-concentrating cells for evaluation of EMT markers and CSCs. This type of study also will reveal the effect of butein on iodine uptake and redifferentiation in advanced metastatic thyroid cancer cells. It was already known that quercetin treatment increases NIS expression in NPA cells and in anaplastic thyroid cancer cells, iodine-131 labeled quercetin was accumulated more in comparison to iodine 131 alone [35,36]. For long-term exposure studies, it is also important to study the effect of butein when used in combination with EBRT and chemotherapy to increase the efficacy of these therapies.

The most important aspect of any novel chemotherapeutics is not only the anti-cancerous activity but also its toxicity towards normal cells. Chemotherapy, radiotherapy, and various protein receptors or protein kinase inhibitors are associated with strong side effects. Hence, toxicological aspects of butein are important from a therapy point of view. Toxicological studies have shown that butein is safe for the normal cells and an oral administration of butein into the rats was found to be safe. Butein had no cytotoxicity towards monocytes, normal lymphocytes, and oral cells like periodontal ligament cells, pulp cells, and gingival fibroblast cells [37]. However, its effects on normal thyrocytes, hormone synthesis by thyroid, and its metabolism are not known. Hence, the effect of butein needs to be investigated in a proper animal thyroid model to explore the effect on the normal thyroid gland.

In summary, the progression and promotion of resistance to therapies, resulting from EMT activation and CSCs emergence has led to the importance of EMT and CSCs promoting signaling pathways in thyroid cancer. Further investigation in the mechanisms by which butein inhibits EMT and CSCs in thyroid cancer will contribute to the mechanistic aspect involved especially when the use of butein is considered as a thyroid cancer therapeutic agent.


1. 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: a Cancer Journal for Clinicians. 2021 May;71(3):209-49.

2. Fullmer T, Cabanillas ME, Zafereo M. Novel therapeutics in radioactive iodine-resistant thyroid cancer. Frontiers in Endocrinology. 2021:836.

3. Nervo A, Ragni A, Retta F, Gallo M, Piovesan A, Liberini V, Gatti M, et al. Bone metastases from differentiated thyroid carcinoma: current knowledge and open issues. Journal of Endocrinological Investigation. 2021 Mar;44(3):403-19.

4. Shibue T, Weinberg RA. EMT, CSCs, and drug resistance: the mechanistic link and clinical implications. Nature reviews Clinical Oncology. 2017 Oct;14(10):611-29.

5. Liskova A, Koklesova L, Samec M, Smejkal K, Samuel SM, Varghese E, et al. Flavonoids in cancer metastasis. Cancers. 2020 Jun 8;12(6):1498.

6. Meerson A, Khatib S, Mahajna J. Flavonoids Targeting Cancer Stem Cells for Augmenting Cancer Therapeutics. International Journal of Molecular Sciences. 2021 Dec 2;22(23):13044.

7. Salehi B, Quispe C, Chamkhi I, El Omari N, Balahbib A, Sharifi-Rad J, et al. Pharmacological properties of chalcones: a review of preclinical including molecular mechanisms and clinical evidence. Frontiers in Pharmacology. 2021 Jan 18;11:592654.

8. Padmavathi G, Roy NK, Bordoloi D, Arfuso F, Mishra S, Sethi G, et al. Butein in health and disease: A comprehensive review. Phytomedicine. 2017 Feb 15;25:118-27.

9. Tripathi D, Kulkarni S. Butein induces intrinsic pathway of apoptosis, vimentin proteolysis, and inhibition of cancer stem cell population in a human papillary thyroid cancer cell line. Toxicology in Vitro. 2021 Dec 1;77:105244.

10. Bayet-Robert M, Morvan D. Metabolomics reveals metabolic targets and biphasic responses in breast cancer cells treated by curcumin alone and in association with docetaxel. PloS One. 2013 Mar 5;8(3):e57971.

11. Yang YC, Lii CK, Lin AH, Yeh YW, Yao HT, Li CC, et al. Induction of glutathione synthesis and heme oxygenase 1 by the flavonoids butein and phloretin is mediated through the ERK/Nrf2 pathway and protects against oxidative stress. Free Radical Biology and Medicine. 2011 Dec 1;51(11):2073-81.

12. Lee DS, Li B, Kim KS, Jeong GS, Kim EC, Kim YC. Butein protects human dental pulp cells from hydrogen peroxide-induced oxidative toxicity via Nrf2 pathway-dependent heme oxygenase-1 expressions. Toxicology In Vitro. 2013 Mar 1;27(2):874-81.

13. Arun R, Dhivya S, Abraham SK, Premkumar K. Low-dose chemotherapeutic drugs induce reactive oxygen species and initiate apoptosis-mediated genomic instability. Toxicology Research. 2016 Mar 1;5(2):547-56.

14. Shah PP, Dupre TV, Siskind LJ, Beverly LJ. Common cytotoxic chemotherapeutics induce epithelial-mesenchymal transition (EMT) downstream of ER stress. Oncotarget. 2017 Apr 4;8(14):22625.

15. Phi LT, Sari IN, Yang YG, Lee SH, Jun N, Kim KS, Lee YK, Kwon HY. Cancer stem cells (CSCs) in drug resistance and their therapeutic implications in cancer treatment. Stem cells international. 2018 Oct;2018.

16. Shibue T, Weinberg RA. EMT, CSCs, and drug resistance: the mechanistic link and clinical implications. Nature reviews Clinical Oncology. 2017 Oct;14(10):611-29.

17. Calangiu CM, Simionescu CE, Stepan AE, Cernea D, Zăvoi RE, Mărgăritescu CL. The expression of CK19, vimentin and E-cadherin in differentiated thyroid carcinomas. Rom J Morphol Embryol. 2014 Jan 1;55(3):919-25.

18. Battaglia RA, Delic S, Herrmann H, Snider NT. Vimentin on the move: new developments in cell migration. F1000Research. 2018;7.

19. Gan Z, Ding L, Burckhardt CJ, Lowery J, Zaritsky A, Sitterley K, et al. Vimentin intermediate filaments template microtubule networks to enhance persistence in cell polarity and directed migration. Cell Systems. 2016 Sep 28;3(3):252-63.

20. Ma R, Bonnefond S, Morshed SA, Latif R, Davies TF. Stemness is derived from thyroid cancer cells. Frontiers in Endocrinology. 2014 Jul 15;5:114.

21. Eriksson JE, He T, Trejo-Skalli AV, Härmälä-Braskén AS, Hellman J, Chou YH, et al. Specific in vivo phosphorylation sites determine the assembly dynamics of vimentin intermediate filaments. Journal of Cell Science. 2004 Feb 22;117(6):919-32.

22. Helfand BT, Mendez MG, Murthy SP, Shumaker DK, Grin B, Mahammad S, et al. Vimentin organization modulates the formation of lamellipodia. Molecular Biology of the Cell. 2011 Apr 15;22(8):1274-89.

23. Goto H, Yasui Y, Kawajiri A, Nigg EA, Terada Y, Tatsuka M, et al. Aurora-B regulates the cleavage furrow-specific vimentin phosphorylation in the cytokinetic process. Journal of Biological Chemistry. 2003 Mar 7;278(10):8526-30.

24. Ivaska J, Vuoriluoto K, Huovinen T, Izawa I, Inagaki M, Parker PJ. PKCε‐mediated phosphorylation of vimentin controls integrin recycling and motility. The EMBO journal. 2005 Nov 16;24(22):3834-45.

25. Zhu QS, Rosenblatt K, Huang KL, Lahat G, Brobey R, Bolshakov S, et al. Vimentin is a novel AKT1 target mediating motility and invasion. Oncogene. 2011 Jan;30(4):457-70.

26. Bollong MJ, Pietilä M, Pearson AD, Sarkar TR, Ahmad I, Soundararajan R, et al. A vimentin binding small molecule leads to mitotic disruption in mesenchymal cancers. Proceedings of the National Academy of Sciences. 2017 Nov 14;114(46):E9903-12.

27. Hammoudi N, Ahmed KB, Garcia-Prieto C, Huang P. Metabolic alterations in cancer cells and therapeutic implications. Chinese Journal of Cancer. 2011 Aug;30(8):508.

28. Sciacovelli M, Frezza C. Metabolic reprogramming and epithelial‐to‐mesenchymal transition in cancer. The FEBS Journal. 2017 Oct;284(19):3132-44.

29. Mitsiades CS, Kotoula V, Poulaki V, Sozopoulos E, Negri J, Charalambous E, et al. Epidermal growth factor receptor as a therapeutic target in human thyroid carcinoma: mutational and functional analysis. The Journal of Clinical Endocrinology & Metabolism. 2006 Sep 1;91(9):3662-6.

30. Sigismund S, Avanzato D, Lanzetti L. Emerging functions of the EGFR in cancer. Molecular Oncology. 2018 Jan;12(1):3-20.

31. Liao W, Liu J, Zhang D, Huang W, Chen R. Butein inhibited in vitro hexokinase-2-mediated tumor glycolysis in hepatocellular carcinoma by blocking epidermal growth factor receptor (EGFR). Medical Science Monitor: International Medical Journal of Experimental and Clinical Research. 2018;24:3283.

32. Zhang C, Zong X, Han Y. Effect of butein and glucose on oxidative stress and p38 activation marker in non-small cell lung cancer cell. Human & Experimental Toxicology. 2019 Oct;38(10):1155-67.

33. Suzuki K, Mitsutake N, Saenko V, Suzuki M, Matsuse M, Ohtsuru A, et al. Dedifferentiation of human primary thyrocytes into multilineage progenitor cells without gene introduction. PLoS One. 2011 Apr 27;6(4):e19354.

34. Yu XM, Jaskula-Sztul R, Ahmed K, Harrison AD, Kunnimalaiyaan M, Chen H. Resveratrol Induces Differentiation Markers Expression in Anaplastic Thyroid Carcinoma via Activation of Notch1 Signaling and Suppresses Cell GrowthResveratrol Induces Redifferentiation in ATC through Notch1. Molecular Cancer Therapeutics. 2013 Jul 1;12(7):1276-87.

35. Kang HJ, Youn YK, Hong MK, Kim LS. Antiproliferation and redifferentiation in thyroid cancer cell lines by polyphenol phytochemicals. Journal of Korean Medical Science. 2011 Jul 1;26(7):893-9.

36. Xie Q, Li X, Wang G, Hou X, Wang Y, Yu H, et al. Preparation and evaluation of 131I-quercetin as a novel radiotherapy agent against dedifferentiated thyroid cancer. Journal of Radioanalytical and Nuclear Chemistry. 2017 Mar;311(3):1697-708.

37. Semwal RB, Semwal DK, Combrinck S, Viljoen A. Butein: From ancient traditional remedy to modern nutraceutical. Phytochemistry Letters. 2015 Mar 1;11:188-201.

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