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Apoptotic morphology of A549 cells treated with bufalin. A549 cells were treated with vehicle (a) or 100 nmol/L bufalin (b) for 48 h and stained by Giemsa staining. cytoplasmic shrinkage, nuclear condensation and the formation of apoptotic bodies were observed in (b). Shown are representative images of three independent experiments. Scale bar: 40 μm.
Source publication
Bufalin is a class of toxic steroids which could induce the differentiation and apoptosis of leukemia cells, and induce the apoptosis of gastric, colon and breast cancer cells. However, the anti-tumor effects of bufalin have not been demonstrated in lung cancer. In this study we used A549 human lung adenocarcinoma epithelial cell line as the experi...
Context in source publication
Context 1
... confirm that bufalin induces the morphology of apoptosis in A549 cells, we treated A549 cells with 100 nmol/L bufalin for 48 h and then performed Giemsa staining. The apoptotic morphology was obviously observed in bufalin treated cells under a microscope at 200× magnification, including cytoplasmic shrinkage, nuclear condensation and the formation of apoptotic bodies (Figure 2). However, no apoptotic morphology was observed in control treated cells. ...Similar publications
Cholangiocarcinoma (CCA) is an aggressive disease with limited effective treatment options. The PI3K/Akt/mTOR pathway represents an attractive therapeutic target due to its frequent dysregulation in CCA. MK2206, an allosteric Akt inhibitor, has been shown to reduce cellular proliferation in other cancers. We hypothesized that MK2206 mediated inhibi...
The prognosis of gastric cancer (GC) remains poor due to clinical drug resistance, and novel drugs are urgently needed. Apoptin-derived peptide (AdP) is an antitumor polypeptide constructed in our laboratory that has been used to combat cisplatin (CDDP) resistance in GC cells. MTT and colony-formation assays and Hoechst 33342 staining were used to...
Background and objective:
In the clinical treatment of patients with non-small cell lung cancer (NSCLC), the primary and acquired resistance of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) limits its clinical application, this need to explore new strategy or method to overcome this problem. Recently, some literatures have...
Background
Tubeimoside-1 (TBMS1) is a natural compound isolated from tubeimoside, which has been widely used as a traditional Chinese herbal medicine. The purpose of the present study is to investigate the anti-tumor effect and the underling mechanism of TBMS1 on glioma cancer cells.
Methods
The MTT assay was performed to evaluate the effect of TB...
Citations
... In this review, we would like to have an overview of bufalin, the most studied bufadienolide, in treating refractory and resistant cancers. Bufalin has been shown significant therapeutic effects in lung cancer (Zhu et al., 2012), bladder cancer (Hong and Choi, 2012), breast cancer (Yan et al., 2012a), oral cancer (Tsai et al., 2012), colon cancer (Xie et al., 2011), gastric cancer (Li et al., 2009), ovarian cancer (Takai et al., 2008), suggesting it is a broad-spectrum anticancer agent. Of note, bufalin has been extensively studied after 2012, especially in drug-resistant or refractory cancers. ...
Profound progress has been made in cancer treatment in the past three decades. However, drug resistance remains prevalent and a critical challenge. Drug resistance can be attributed to oncogenes mutations, activated defensive mechanisms, ATP-bind cassette transporters overexpression, cancer stem cells, etc. Chinese traditional medicine toad venom has been used for centuries for different diseases, including resistant cancers. Bufalin is one of the bufadienolides in toad venom that has been extensively studied for its potential in refractory and drug-resistant cancer treatments in vitro and in vivo . In this work, we would like to critically review the progress made in the past decade (2013–2022) of bufalin in overcoming drug resistance in cancers. Generally, bufalin shows high potential in killing certain refractory and resistant cancer cells via multiple mechanisms. More importantly, bufalin can work as a chemo-sensitizer that enhances the sensitivity of certain conventional and targeted therapies at low concentrations. In addition, the development of bufalin derivatives was also briefly summarized and discussed. We also analyzed the obstacles and challenges and provided possible solutions for future perspectives. We hope that the collective information may help evoke more effort for more in-depth studies and evaluation of bufalin in both lab and possible clinical trials.
... sensitive and/or resistant to commercially available drugs in vitro and in vivo [1,2,9]. Mechanistic studies have shown that bufadienolides induce various cell death mechanisms such as apoptosis, autophagy and ferroptosis [10][11][12][13]. They also act as antiangiogenic and immunosuppressants [14]. ...
... The cytotoxic effect of bufadienolides on A549 cells has been extensively studied [6,7,10,13,50]. Bufotalin induces ferroptosis in A549 cells by accelerating the degradation of GPX4 and raising the intracellular Fe 2+ ; however, the IC 50 value for cell viability/metabolism [13] is orders of magnitude higher than that obtained in the present study; the reason for this discrepancy is unknown. Other bufadienolides also induce cell death in A549 cells through apoptosis [10][11][12]. ...
... Bufotalin induces ferroptosis in A549 cells by accelerating the degradation of GPX4 and raising the intracellular Fe 2+ ; however, the IC 50 value for cell viability/metabolism [13] is orders of magnitude higher than that obtained in the present study; the reason for this discrepancy is unknown. Other bufadienolides also induce cell death in A549 cells through apoptosis [10][11][12]. On the other hand, the combination of bufalin with sorafenib results in a greater cytotoxic effect on cell viability/metabolism and cell death in the lung cell line H292 [52]; these results are the consequence of enhanced chromatin condensation and increased apoptosis, compared to individual treatments. ...
Bufadienolides are steroids that inhibit the Na ⁺ /K ⁺ ATPase pump. Recent studies show that members of the bufadienolide family also inhibit the activity of aminopeptidase N (APN). APN is upregulated in different pathologies, including cancer and is a current target for drug development. Bufadienolides are cytotoxic in tumor cells, but there is no enough evidences that inhibition of APN activity contributes to their effect. In the present contribution we investigated the effect of another member of the bufadienolide family, bufotalin, on porcine APN (pAPN) activity. Bufotalin inhibited pAPN activity with K i values in the submicromolar range and an uncompetitive inhibition mechanism; it also inhibited porcine aminopeptidase A (pAPA) activity, but with a classical reversible competitive inhibition mechanism. In addition, we determined the effect of bufotalin on the viability/metabolism of APN+ A549, H292, MeWo and CT26 cancer cells. Bufotalin was cytotoxic in a dose dependent manner; the highest cytotoxicity was detected in A549 cells, the cells with the highest APN activity. Thus, tumor cell line sensitivity to the cytotoxic effect of bufotalin correlates with cell surface APN activity.
... In the aspect of antitumor, Bufalin can inhibit the growth of tumor cells, promote the differentiation of tumor cells and induce the apoptosis of tumor cells (53). Bufalin can inhibit tumor cell proliferation by blocking phosphatidylinositol-3kinase/serine protein kinase signal pathway, and induce tumor cell apoptosis through mitochondrial or death receptor-mediated pathway (54), which can reverse the drug resistance of hepatocellular carcinoma cells to fluorouracil (22 (56). It has been found that Bufalin can effectively inhibit the metastasis of lung cancer NCI-H460 cells, which may be related to the inhibition of matrix metalloproteinase-9 (MMP-9) activity, the decrease of cytoplasmic NF-κB production and the reduction of NF-κB nuclear translocation (57). ...
Background:
Esophageal cancer, especially esophageal squamous cell carcinoma (ESCC), is a common malignant tumor of the digestive tract. Bufalin is an effective anti-tumor compound. However, little is known about the regulatory mechanisms of Bufalin in ESCC. To investigate the effect and molecular mechanism of Bufalin on the proliferation, migration and invasion of ESCC cells will provide a more reliable basis for the application of Bufalin in clinical tumor therapy.
Methods:
First, the half-inhibitory concentration (IC50) of Bufalin was evaluated by Cell Counting Kit-8 (CCK-8) assays. In vitro, the effects of Bufalin on the proliferation of the ECA109 cells was measured using CCK-8 and 5-ethynyl-2'-deoxyuridine assays. Wound-healing and transwell assays were used to evaluate the effects of Bufalin on the migration and invasion of the ECA109 cells. Further, to determine the mechanisms underlying the Bufalin-mediated suppression of cell progression in ESCC, total RNA was extracted from negative control (NC) and Bufalin treated cells to perform RNA-sequencing (RNA-seq) to screen for abnormally expressed genes. In vivo, the ECA 109 cells were subcutaneously injected into BALB/c nude mice to determine the effects of Bufalin on tumor cell proliferation. The protein inhibitor of activated signal transducer and activator of transcription 3 (PIAS3), signal transducer and activator of transcription 3 (STAT3), and phosphorylated STAT3 (p-STAT3) protein expression levels in the ECA109 cells were detected by Western blot.
Results:
The CCK-8 assays showed that the IC50 of Bufalin was 200 nM. The proliferation, migration, and invasion ability of the ECA109 cells was significantly inhibited in the Bufalin group in a concentration-dependent manner. In vivo, the Xenograft tumor model showed that Bufalin decreased the tumor volume and weight of the subcutaneous tumors. The RNA-seq results showed that the expression of PIAS3 was upregulated in the Bufalin group. Additionally, down-regulation of PIAS3 decreased the inhibition of STAT3, thereby increasing p-STAT3 expression. Finally, PIAS3 knockdown reversed the inhibitory effects of Bufalin on the proliferation, migration, and invasion of the ECA109 cells.
Conclusions:
Bufalin may inhibit the proliferation, migration, and invasion of the ECA109 cells through the PIAS3/STAT3 signaling pathway.
... Besides, cinobufacini injection also effectively protects liver function in moderate and advanced primary liver cancer. Experimental studies have shown that bufalin exerts antitumor activities in various cancer cells including HL-60, A549 and HCT116 [25][26][27] . In particular, bufalin induces apoptosis and inhibits migration and invasion in human HCC cell lines [28][29][30][31] . ...
Syndecan-4 (SDC4) functions as a major endogenous membrane-associated receptor and widely regulates cytoskeleton, cell adhesion, and cell migration in human tumorigenesis and development, which represents a charming anti-cancer therapeutic target. Here, SDC4 was identified as a direct cellular target of small-molecule bufalin with anti-hepatocellular carcinoma (HCC) activity. Mechanism studies revealed that bufalin directly bond to SDC4 and selectively increased SDC4 interaction with substrate protein DEAD-box helicase 23 (DDX23) to induce HCC genomic instability. Meanwhile, pharmacological promotion of SDC4/DDX23 complex formation also inactivated matrix metalloproteinases (MMPs) and augmented p38/JNK MAPKs phosphorylation, which are highly associated with HCC proliferation and migration. Notably, specific knockdown of SDC4 or DDX23 markedly abolished bufalin-dependent inhibition of HCC proliferation and migration, indicating SDC4/DDX23 signaling axis is highly involved in the HCC process. Our results indicate that membrane-spanning proteoglycan SDC4 is a promising druggable target for HCC, and pharmacological regulation of SDC4/DDX23 signaling axis with small-molecule holds great potential to benefit HCC patients.
... The mixed methods research that will connect the active substance, signaling pathway and apoptotic death of cancer cells is the most important. Bufalin and PI3K / Akt Pathway, or Sesamin and Akt / p53 pathway in the induction of apoptosis are examples of multi-method works that give the complete picture of the subject of research [53,54]. ...
Epidemiological data indicate that in the last period there has been an increase in the number of
malignancies and among them lung cancer is one of the most common forms. In vitro studies based on the usage of
immortalized cell lines are an important source of scientific knowledge for understanding of the mechanism of cell
growth, proliferation and cell death. In this paper, the most commonly used methods for in vitro research in NSCLC
(non-small cell lung cancer) based on testing the effects of new compounds to determine the degree of apoptosis,
necrosis, cell proliferation as well as their significance are discussed. So far, techniques of working with monolayer
cultures have been mainly used. In the future, it is recommended to use a 3D system, knockout cell line and to conduct
additional studies regarding the use of organoids or spheroids, as well as the application of new techniques to better
understand the complex processes of carcinogenesis and the action of biologically active compounds.
... AKT1 is involved in various biological processes, including cell metabolism, survival, and growth, and its abnormal expression correlated with poor prognosis of tumor patients [50,51]. Zhu et al. [52] found that bufalin, the main compound of cinobufotalin injection, induces apoptosis of lung cancer cells in a dose-and time-dependent manner by upregulating Bax expression and downregulating BCL-2, thereby blocking the activation of AKT. We found that in lung squamous cell carcinoma, CCND1, the main potential therapeutic target of cinobufotalin injection, was associated with patient survival. ...
Cinobufotalin injection, extracted from the skin of Chinese giant salamander or black sable, has good clinical effect against lung cancer. However, owing to its complex composition, the pharmacological mechanism of cinobufotalin injection has not been fully clarified. This study aimed to explore the mechanism of action of cinobufotalin injection against lung cancer using network pharmacology and bioinformatics. Compounds of cinobufotalin injection were determined by literature retrieval, and potential therapeutic targets of cinobufotalin injection were screened from Swiss Target Prediction and STITCH databases. Lung-cancer-related genes were summarized from GeneCards, OMIM, and DrugBank databases. The pharmacological mechanism of cinobufotalin injection against lung cancer was determined by enrichment analysis of gene ontology and Kyoto Encyclopedia of Genes and Genomes, and protein-protein interaction network was constructed. We identified 23 compounds and 506 potential therapeutic targets of cinobufotalin injection, as well as 70 genes as potential therapeutic targets of cinobufotalin injection in lung cancer by molecular docking. The antilung cancer effect of cinobufotalin injection was shown to involve cell cycle, cell proliferation, antiangiogenesis effect, and immune inflammation pathways, such as PI3K-Akt, VEGF, and the Toll-like receptor signaling pathway. In network analysis, the hub targets of cinobufotalin injection against lung cancer were identified as VEGFA, EGFR, CCND1, CASP3, and AKT1. A network diagram of “drug-compounds-target-pathway” was constructed through network pharmacology to elucidate the pharmacological mechanism of the antilung cancer effect of cinobufotalin injection, which is conducive to guiding clinical medication.
... Bufalin, one of the major active components in VB, is regarded as a vital marker compound for the quality control of VB, along with its related TCM preparations [5]. Arrays of studies have shown its powerful cytotoxic activities against various tumor cells, such as human leukemia cells, lung cancer cells, human hepatocellular carcinoma HepG2 cells, human malignant melanoma A375.S2 cells, etc. [6][7][8]. Meanwhile, bufalin possesses powerful cardiotoxicity and neurotoxicity, and it can result in convulsion, diarrhea, arrhythmia, and other symptoms [9,10]. ...
Characterization and determination of metabolites to monitor metabolic pathways play a paramount role in evaluating the efficacy and safety of medicines. However, the separation and quantification of metabolites are rather difficult due to their limited contents in vivo, especially in the case of Chinese medicine, due to its complexity. In this study, an effective and convenient method was developed to simultaneously quantify bufalin and its nine metabolites (semi-quantitation) in rat plasma after an oral administration of 10 mg/kg to rats. The prototype and metabolites that were identified were subsequently quantified using positive electrospray ionization in multiple reaction monitoring (MRM) mode with transitions of m/z 387.4→369.6 and 387.4→351.3 for bufalin, m/z 513.7→145.3 for IS, and 387.4→369.6, 419.2→365.2, and 403.2→349.2 for the main metabolites (3-epi-bufalin, dihydroxylated bufalin, and hydroxylated bufalin, respectively). The method was validated over the calibration curve range of 1.00–100 ng/mL with a limit of quantitation (LOQ) of 1 ng/mL for bufalin. No obvious matrix effect was observed, and the intra- and inter-day precisions, as well as accuracy, were all within the acceptable criteria in this method. Then, this method was successfully applied in metabolic profiling and a pharmacokinetic study of bufalin after an oral administration of 10 mg/kg to rats. The method of simultaneous determination of bufalin and its nine metabolites in rat plasma could be useful for pharmacokinetic–pharmacodynamic relationship research of bufalin, providing experimental evidence for explaining the occurrence of some adverse effects of Venenum Bufonis and its related preparations.
... For example, oleandrin induces apoptosis by the activation of caspases and upregulation of Bax expression in colon cancer cells [13]. In addition, digoxin, bufalin, and ouabain have been reported to inhibit cell growth by regulating multiple signaling pathways including topoisomerases I and II [15], hypoxia-inducible factor 1 [16], PI3K/Akt [17], and Bcl-2/Bax [18]. Furthermore, PBI-0524 and Anvirzel have been evaluated for their antitumor potency in clinical trials for the treatment of solid tumors and have been proven to be safe and effective [19,20]. ...
Cardiac glycosides (CGs), prescribed to treat congestive heart failure and arrhythmias, exert potent antitumor activity. In this study, divaricoside (DIV), a CG isolated from Strophanthus divaricatus was examined for its antitumor potency in oral squamous cell carcinoma (OSCC) cells. Cell growth was inhibited by DIV in a dose- and time-dependent manner in SCC2095 and OECM-1 OSCC cells using MTT assays. DIV induced S and G2/M phase arrest accompanied by downregulation of phosphorylated CDC25C, CDC25C, and CDC2 in SCC2095 cells. In addition, DIV induced apoptosis by activating caspase-3 and downregulating the expression of Mcl-1. Furthermore, overexpression of Mcl-1 partially reversed DIV-induced death in SCC2095 cells. Additionally, western blot and transmission electron microscopy analyses also indicated that DIV induced autophagy in SCC2095 cells. However, the combination of autophagy inhibitor did not affect DIV-mediated apoptosis in SCC2095 cells. Together, these findings suggest that translational potential of DIV to be developed as a therapeutic agent for OSCC treatment.
... Owing to the similarity in the chemical structure between bufalin and digoxin, bufalin is expected to have a digoxin-like function (5,6). Bufalin has been shown to induce apoptosis in leukemia and solid tumor cells (5)(6)(7)(8)(9)(10). In recent years, studies have revealed that bufalin could effectively inhibit the proliferation of tumor cells, and also assessed its possible molecular mechanism (6)(7)(8). ...
... Bufalin has been shown to induce apoptosis in leukemia and solid tumor cells (5)(6)(7)(8)(9)(10). In recent years, studies have revealed that bufalin could effectively inhibit the proliferation of tumor cells, and also assessed its possible molecular mechanism (6)(7)(8). The results of previous research also demonstrated that bufalin could inhibit the activation of ERK pathway, accordingly inhibit the proliferation and migration of esophageal carcinoma cells (11); however, the mechanism of inducing apoptosis has remained unclear. ...
The present study examined whether bufalin could induce human esophageal carcinoma ECA109 cells apoptosis via inhibiting the activation of mechanistic target of rapamycin (mTOR)/p70 S6 kinase (p70S6K) pathway is discussed in this article. The present study used the esophageal squamous cell carcinoma ECA109 cell line to assess the apoptosis-inducing effects of bufalin via inhibition of the mTOR/p70S6K pathways. A plasmid containing the wild-type mTOR gene (wtmTOR) was transfected into ECA109 cells. The levels of p70S6K, phosphorylated (p)-p70S6K, cellular inhibitor of apoptosis-1 (cIAP-1) and Bcl-2-associated death promoter (BAD) in ECA109 cells were examined by western blot analysis, and apoptosis was detected by flow cytometry analysis and Giemsa staining. The results revealed that the expression of p-p70S6K was increased as the time progressed (at 0, 12 and 24 h), and then decreased at 30, 36, 42 and 48 h after transfection. The expression of cIAP-1 was significantly decreased as time progressed following the addition of bufalin, whereas that of BAD was increased. The levels of p-p70S6K and cIAP-1 were significantly higher in the wtmTOR-transfected group than in the control and empty vector-transfected groups, and then reduced following addition of bufalin; however, BAD expression was significantly lower in the wtmTOR-transfected group. The results of flow cytometry revealed the cell cycle of ECA109 was arrested at G2/M phase and the apoptotic rate was significantly lower in the wtmTOR-transfected group than in the control and empty vector-transfected groups, and then increased following addition of bufalin. In conclusion, the findings of the present study demonstrated that bufalin induced apoptosis in esophageal carcinoma cells via the inhibition of the mTOR/p70S6K pathway and indicated that treatment with bufalin could be combined with chemotherapy to overcome the resistance of esophageal carcinoma cells to chemotherapeutic-induced apoptosis.
... In recent years, bufalin have been reported to exert significant anti-cancer effects in a variety of tumor cells, including gynecologic cancers [17], HCC cells [18], lung cancer [19,20] , breast cancer [21], colon cancer [22], and osteosarcoma [23]. The mechanisms underlying bufalin induction of cancer cell apoptosis are documented to be correlated with upregulation of Bax, downregulation of Bcl-2, activation of caspase cascade, and inhibition of PI3K/AKT pathway [19,24,25]. Recently, we determined that bufalin could induce apoptosis in NSCLC cells by down-regulating Mcl-1 [19]. ...
