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ONCOLOGY / BASIC RESEARCH
Cinobufagin disrupts the stability of lipid rafts by inhibiting the expression of caveolin-1 to promote non-small cell lung cancer cell apoptosis
1
Department of Gerontology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
Submission date: 2023-08-02
Final revision date: 2023-10-25
Acceptance date: 2023-10-27
Online publication date: 2024-06-28
Corresponding author
Arch Med Sci 2024;20(3):887-908
KEYWORDS
TOPICS
ABSTRACT
Introduction:
The study was designed to explore how cinobufagin (CB) regulates the development of non-small cell lung cancer (NSCLC) cells through lipid rafts.
Material and methods:
The effects of CB at gradient concentrations (0, 0.5, 1 and 2 µM) on NSCLC cell viability, apoptosis, reactive oxygen species (ROS) level, phosphorylation of Akt, and apoptosis- and lipid raft-related protein expression were assessed by MTT assay, flow cytometry and Western blot. Cholesterol and sphingomyelin were labeled with BODIPY to evaluate the effect of CB (2 µM) on them. Sucrose density gradient centrifugation was used to extract lipid rafts. The effect of CB on the expression and distribution of caveolin-1 was determined by immunofluorescence, quantitative reverse transcription polymerase chain reaction and Western blot. After overexpression of caveolin-1, the above experiments were performed again to observe whether the regulatory effect of CB was reversed.
Results:
CB inhibited NSCLC cell viability while promoting apoptosis and ROS level. CB redistributed the lipid content on the membrane surface and reduced the content of caveolin-1 in the cell membrane. In addition, CB repressed the activation of AKT. However, caveolin-1 overexpression reversed the effects of CB on apoptosis, AKT activation and lipid raft.
Conclusions:
CB regulates the activity of Akt in lipid rafts by inhibiting caveolin-1 expression to promote NSCLC cell apoptosis.
The study was designed to explore how cinobufagin (CB) regulates the development of non-small cell lung cancer (NSCLC) cells through lipid rafts.
Material and methods:
The effects of CB at gradient concentrations (0, 0.5, 1 and 2 µM) on NSCLC cell viability, apoptosis, reactive oxygen species (ROS) level, phosphorylation of Akt, and apoptosis- and lipid raft-related protein expression were assessed by MTT assay, flow cytometry and Western blot. Cholesterol and sphingomyelin were labeled with BODIPY to evaluate the effect of CB (2 µM) on them. Sucrose density gradient centrifugation was used to extract lipid rafts. The effect of CB on the expression and distribution of caveolin-1 was determined by immunofluorescence, quantitative reverse transcription polymerase chain reaction and Western blot. After overexpression of caveolin-1, the above experiments were performed again to observe whether the regulatory effect of CB was reversed.
Results:
CB inhibited NSCLC cell viability while promoting apoptosis and ROS level. CB redistributed the lipid content on the membrane surface and reduced the content of caveolin-1 in the cell membrane. In addition, CB repressed the activation of AKT. However, caveolin-1 overexpression reversed the effects of CB on apoptosis, AKT activation and lipid raft.
Conclusions:
CB regulates the activity of Akt in lipid rafts by inhibiting caveolin-1 expression to promote NSCLC cell apoptosis.
REFERENCES (42)
1.
Hirsch FR, Scagliotti GV, Mulshine JL, et al. Lung cancer: current therapies and new targeted treatments. Lancet 2017; 389: 299-311.
2.
Evison M. The current treatment landscape in the UK for stage III NSCLC. Br J Cancer 2020; 123 (Suppl 1): 3-9.
3.
So TH, Chan SK, Lee VH, et al. Chinese medicine in cancer treatment – how is it practised in the East and the West? Clin Oncol (R Coll Radiol) 2019; 31: 578-88.
4.
Liu Y, Yang S, Wang K, et al. Cellular senescence and cancer: focusing on traditional Chinese medicine and natural products. Cell Prolif 2020; 53: e12894.
5.
Li X, Bi Z, Liu S, et al. Antifibrotic mechanism of cinobufagin in bleomycin-induced pulmonary fibrosis in mice. Front Pharmacol 2019; 10: 1021.
6.
Li F J, Hu J H, Ren X, et al. Toad venom: a comprehensive review of chemical constituents, anticancer activities, and mechanisms. Arch Pharm (Weinheim) 2021; 354: e2100060.
7.
Cao Y, Yu L, Dai G, et al. Cinobufagin induces apoptosis of osteosarcoma cells through inactivation of Notch signaling. Eur J Pharmacol 2017; 794: 77-84.
8.
Pan Z, Luo Y, Xia Y, et al. Cinobufagin induces cell cycle arrest at the S phase and promotes apoptosis in nasopharyngeal carcinoma cells. Biomed Pharmacother 2020; 122: 109763.
9.
Kim GH, Fang XQ, Lim WJ, et al. Cinobufagin suppresses melanoma cell growth by inhibiting LEF1. Int J Mol Sci 2020; 21: 6706.
10.
Li X, Chen C, Dai Y, et al. Cinobufagin suppresses colorectal cancer angiogenesis by disrupting the endothelial mammalian target of rapamycin/hypoxia-inducible factor 1 axis. Cancer Sci 2019; 110: 1724-34.
11.
Xie M, Chen X, Qin S, et al. Clinical study on thalidomide combined with cinobufagin to treat lung cancer cachexia. J Cancer Res Ther 2018; 14: 226-32.
12.
Levental I, Levental KR, Heberle FA. Lipid rafts: controversies resolved, mysteries remain. Trends Cell Biol 2020; 30: 341-53.
13.
Sezgin E, Levental I, Mayor S, et al. The mystery of membrane organization: composition, regulation and roles of lipid rafts. Nat Rev Mol Cell Biol 2017; 18: 361-74.
14.
Jeon JH, Kim SK, Kim HJ, et al. Lipid raft modulation inhibits NSCLC cell migration through delocalization of the focal adhesion complex. Lung Cancer 2010; 69: 165-71.
15.
Qu X, Liu Y, Ma Y, et al. Up-regulation of the Cbl family of ubiquitin ligases is involved in ATRA and bufalin-induced cell adhesion but not cell differentiation. Biochem Biophys Res Commun 2008; 367: 183-9.
16.
Zhang G, Wang C, Sun M, et al. Cinobufagin inhibits tumor growth by inducing intrinsic apoptosis through AKT signaling pathway in human nonsmall cell lung cancer cells. Oncotarget 2016; 7: 28935-46.
17.
Dong C, Gongora R, Sosulski ML, et al. Regulation of transforming growth factor-beta1 (TGF-1)-induced pro-fibrotic activities by circadian clock gene BMAL1. Respir Res 2016; 17: 4.
18.
Kumar P, Nagarajan A, Uchil PD. Analysis of cell viability by the MTT assay. Cold Spring Harb Protoc 2018; 2018(6).
19.
Kumar R, Saneja A, Panda AK. An annexin V-FITC-propidium iodide-based method for detecting apoptosis in a non-small cell lung cancer cell line. Methods Mol Biol 2021; 2279: 213-23.
20.
Pillai-Kastoori L, Schutz-Geschwender AR, Harford JA. A systematic approach to quantitative Western blot analysis. Anal Biochem 2020; 593: 113608.
21.
Ding J, Yao Y, Li J, et al. A reactive oxygen species scavenging and O(2) generating injectable hydrogel for myocardial infarction treatment in vivo. Small 2020; 16: e2005038.
22.
Mishra SK, Bae YS, Lee YM, et al. Sesquiterpene alcohol cedrol chemosensitizes human cancer cells and suppresses cell proliferation by destabilizing plasma membrane lipid rafts. Front Cell Dev Biol 2020; 8: 571676.
23.
Niu J, Wang J, Zhang Q, et al. Cinobufagin-induced DNA damage response activates G(2)/M checkpoint and apoptosis to cause selective cytotoxicity in cancer cells. Cancer Cell Int 2021; 21: 446.
24.
Ediriweera MK, Moon JY, Nguyen YT, et al. 10-gingerol targets lipid rafts associated PI3K/Akt signaling in radio-resistant triple negative breast cancer cells. Molecules 2020; 25: 3164.
25.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 2001; 25: 402-8.
26.
Luo X, Cheng C, Tan Z, et al. Emerging roles of lipid metabolism in cancer metastasis. Mol Cancer 2017; 16: 76.
27.
Li B, Qin Y, Yu X, et al. Lipid raft involvement in signal transduction in cancer cell survival, cell death and metastasis. Cell Prolif 2022; 55: e13167.
28.
Badana A, Chintala M, Varikuti G, et al. Lipid raft integrity is required for survival of triple negative breast cancer cells. J Breast Cancer 2016; 19: 372-84.
29.
Ketteler J, Klein D. Caveolin-1, cancer and therapy resistance. Int J Cancer 2018; 143: 2092-104.
30.
Leiser D, Samanta S, Eley J, et al. Role of caveolin-1 as a biomarker for radiation resistance and tumor aggression in lung cancer. PLoS One 2021; 16: e0258951.
31.
Wang Y, Song Y, Che X, et al. Caveolin 1 enhances RANKL induced gastric cancer cell migration. Oncol Rep 2018; 40: 1287-96.
32.
Philips BJ, Kumar A, Burki S, et al. Triptolide-induced apoptosis in non-small cell lung cancer via a novel miR204-5p/Caveolin-1/Akt-mediated pathway. Oncotarget 2020; 11: 2793-806.
33.
Zhao X, Liu Y, Ma Q, et al. Caveolin-1 negatively regulates TRAIL-induced apoptosis in human hepatocarcinoma cells. Biochem Biophys Res Commun 2009; 378: 21-6.
34.
Zinnah KMA, Park SY. Sensitizing TRAIL resistant A549 lung cancer cells and enhancing TRAIL induced apoptosis with the antidepressant amitriptyline. Oncol Rep 2021; 46: 144.
35.
Tan AC. Targeting the PI3K/Akt/mTOR pathway in non-small cell lung cancer (NSCLC). Thorac Cancer 2020; 11: 511-8.
36.
Calay D, Vind-Kezunovic D, Frankart A, et al. Inhibition of Akt signaling by exclusion from lipid rafts in normal and transformed epidermal keratinocytes. J Invest Dermatol 2010; 130: 1136-45.
37.
Pan Z, Zhang X, Yu P, et al. Cinobufagin induces cell cycle arrest at the G2/M phase and promotes apoptosis in malignant melanoma cells. Front Oncol 2019; 9: 853.
38.
Yang AL, Wu Q, Hu ZD, et al. A network pharmacology approach to investigate the anticancer mechanism of cinobufagin against hepatocellular carcinoma via downregulation of EGFR-CDK2 signaling. Toxicol Appl Pharmacol 2021; 431: 115739.
39.
Huang G, Lou T, Pan J, et al. MiR-204 reduces cisplatin resistance in non-small cell lung cancer through suppression of the caveolin-1/AKT/Bad pathway. Aging (Albany NY) 2019; 11: 2138-50.
40.
Li L, Ren CH, Tahir SA, et al. Caveolin-1 maintains activated Akt in prostate cancer cells through scaffolding domain binding site interactions with and inhibition of serine/threonine protein phosphatases PP1 and PP2A. Mol Cell Biol 2003; 23: 9389-404.
41.
Gauthier-Rouvière C, Bodin S, Comunale F, et al. Flotillin membrane domains in cancer. Cancer Metastasis Rev 2020; 39: 361-74.
42.
Zhu M, Shi W, Chen K, et al. Pulsatilla saponin E suppresses viability, migration, invasion and promotes apoptosis of NSCLC cells through negatively regulating Akt/FASN pathway via inhibition of flotillin-2 in lipid raft. J Recept Signal Transduct Res 2022; 42: 23-33.
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