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Therapeutic immunity of tumor lysate-loaded DC vaccines against B16 melanoma. a The vaccination scheme employed in this study. Mice were challenged s.c. with 1 9 10 5 B16 cells and vaccinated with different preparations of tumor cell lysate-loaded DCs. b CTL activities in splenocytes on target tumor cells after vaccination with DCs loaded with SK-TCL, MG-TCL, DX- TCL, or vehicle-TCL (control). PBS-treated mice did not receive any DC vaccines. The mice in the mature DC group were subjected to vaccination with unloaded mature DCs. Tumor growth (c) and survival rates (d) in treated mice. * P \ 0.05, when compared with the control vehicle-TCL. e Typical examples of tumor appearance (in size and morphology) in mice on day 31
Source publication
Immunogenic cell death is characterized by damage-associated molecular patterns, which can enhance the maturation and antigen uptake of dendritic cells. Shikonin, an anti-inflammatory and antitumor phytochemical, was exploited here as an adjuvant for dendritic cell-based cancer vaccines via induction of immunogenic cell death. Shikonin can effectiv...
Contexts in source publication
Context 1
... above mechanistic studies suggest that SK-TCL, DX- TCL or MG-TCL have the potential to effectively activate DC maturation and induce a strong T cell immune response. We therefore investigated possible therapeutic immunities in DX-TCL-, SK-TCL-and MG-TCL-loaded DC-based cancer vaccines against primary B16 melanoma (see schema in Fig. 5a). For mice receiving mature vehicle- TCL-loaded DCs, mature but unloaded DCs, or the PBS control treatment, no significant CTL activity in splenocytes was detected (Fig. 5b). However, the MG-TCL-loaded DC vaccine triggered a good CTL activity that elicited approximately 40 % of specific lysis at effector/target (E:T) ratios ranging ...
Context 2
... response. We therefore investigated possible therapeutic immunities in DX-TCL-, SK-TCL-and MG-TCL-loaded DC-based cancer vaccines against primary B16 melanoma (see schema in Fig. 5a). For mice receiving mature vehicle- TCL-loaded DCs, mature but unloaded DCs, or the PBS control treatment, no significant CTL activity in splenocytes was detected (Fig. 5b). However, the MG-TCL-loaded DC vaccine triggered a good CTL activity that elicited approximately 40 % of specific lysis at effector/target (E:T) ratios ranging from 20:1 to 80:1. CTL activities in spleno- cytes of mice vaccinated with SK-TCL-or DX-TCL-loaded DC vaccines were increased with increasing E/T ratios, reaching statistical ...
Context 3
... activities in spleno- cytes of mice vaccinated with SK-TCL-or DX-TCL-loaded DC vaccines were increased with increasing E/T ratios, reaching statistical significance at the E/T ratio of 80:1. Figure 5c and d show that tumor growth was strongly retarded and survival of mice greatly improved by the use of these two vaccines as compared to PBS control or other reference vaccine sets. The tumor sizes in MG-TCL-treated mice and especially in SK-TCL-and DX-TCL-treated mice were significantly smaller on day 21 (Fig. 5e) as compared to that in mice of the TCL-vehicle control group. ...
Context 4
... at the E/T ratio of 80:1. Figure 5c and d show that tumor growth was strongly retarded and survival of mice greatly improved by the use of these two vaccines as compared to PBS control or other reference vaccine sets. The tumor sizes in MG-TCL-treated mice and especially in SK-TCL-and DX-TCL-treated mice were significantly smaller on day 21 (Fig. 5e) as compared to that in mice of the TCL-vehicle control group. ...
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Citations
... SKN, in synergy with traditional chemotherapeutic drugs, effectively inhibits the growth of liver cancer and enhances the efficacy of specific drugs while reducing drug resistance [15][16] . Furthermore, SKN is a potent inducer of ICD, enhancing cellular oxidative stress by activating mitochondrion-and receptor-mediated apoptosis pathways, thereby promoting DC-based immunotherapy effectiveness [17][18] . Hence, SKN is extensively used in tumor immunotherapy, particularly as an adjuvant for DC vaccines. ...
... Furthermore, shikonin has demonstrated synergistic effects when combined with conventional cancer treatments, such as chemotherapy, immunotherapy, and radiotherapy [34]. In addition to its cytotoxic properties, shikonin shows promise to be used for the development of cancer vaccines, enhancing the tumor-specific immune response against cancerous cells [35]. Despite the substantial body of research papers investigating the anti-BRCA effects of shikonin, a wide-ranging overview of the available studies on its applications against BRCA has not yet been conducted. ...
Objectives
Breast cancer is a prevalent disease that has a substantial impact on women’s mortality rates. Shikonin, a naphthoquinone derived from Lithospermum erythrorhizon, has demonstrated substantial anticancer effects. This study aims to conduct a comprehensive review of the latest research findings regarding the therapeutic efficacy of shikonin in the context of breast cancer treatment, with a specific emphasis on elucidating the underlying molecular mechanisms.
Methods
A comprehensive literature review was conducted on shikonin and breast cancer by searching PubMed, Scopus, Web of Science, and Google Scholar databases.
Key findings
Shikonin significantly reduces tumor cell viability, proliferation, migration, invasion, and metastasis in both in vivo and in vitro across all breast cancer subtypes. Additionally, when combined with other pharmaceutical agents, it exhibits synergistic effects. Shikonin stimulates immunogenic cell death, resulting in apoptosis and necroptosis. The induction of immunogenic cell death by shikonin enhances the immunogenicity of breast cancer cells, leading to its involvement in the development of dendritic cell-based tumor vaccines against breast cancer.
Conclusion
Shikonin exhibits potent anti-breast cancer properties and shows significant potential for the advancement of immunotherapeutic approaches against breast cancer, as well as enhancing the efficacy of conventional treatment strategies.
... However, the complete understanding of the mechanism behind Hsp70 upregulation induced by Shikonin is still under investigation. Chen et al. has found that SK could induce ICD of melanoma B16 cells and enhance function of dendritic cells (DCs) [19]. Previous studies have confirmed that SK and BET inhibitor JQ1 can synergistically exert anti-tumor effects by reshaping the tumor immune microenvironment [20]. ...
... Tumor cell lysate samples were prepared as previously described [19]. The protein samples were resolved by SDS-PAGE using 10 or 15% stepwise gels. ...
... Chen et al. concluded that Shikonin could effectively increase the production of specific DAMPs in B16 cells, including Hsp70, Hsp90, CRT and HMGB1. Shikonin-treated B16 cell lysates could induce DCs to a higher level of functional and phenotypic maturation, exhibiting high expression of CD86 and MHC class II and Th1 cells activation [19]. In our study, we also found that the percentage of CD11 + CD11b + DC and CD8 + T cells have been increased, suggesting that synergistic effect of SK with PD-1 blockade in CT26 tumor mice model might due to DC activation and cytotoxic CD8 + T cells infiltration. ...
Background
PD-1 blockade has shown impressive clinical outcomes in colorectal cancers patients with high microsatellite instability (MSI-H). However, the majority of patients with colorectal cancer who present low microsatellite instability (MSI-L) or stable microsatellites (MSS) show little response to PD-1 blockade therapy. Here, we have demonstrated that Shikonin (SK) could induce cell death of CT26 cells via classically programmed and immunogenic pathways.
Methods and results
SK promoted the membrane exposure of calreticulin and upregulated the expression of heat shock protein 70 (Hsp70). The upregulation of Hsp70 was dependent on ROS induced by SK and silencing of PKM2 in CT26 cells reverts ROS upregulation. Besides, SK synergizes with PD-1 blockade in CT26 tumor mice model, with the increase of intramural DC cells and CD8+ T cells. The expression of Hsp70 in tumor tissue was also increased in combinational SK plus αPD-1 therapy group.
Conclusions
Our study elucidated the potential role of ‘Shikonin-PKM2-ROS-Hsp70’ axis in the promotion of efficacy of PD-1 blockade in CRC treatments, providing a potential strategy and targets for improving the efficacy of PD-1 blockade in colorectal cancer.
... Further approaches have also been explored to reinforce the effect of anticancer chemotherapy, including the design of systems able to simultaneously carry antineoplastic drugs and bioactive natural compounds known to possess antioxidant, anti-inflammatory, and anticancer activities themselves, such as PheCs, omega-3 fatty acids, metabolites derived from garlic or Brassicaceae, or many others [20][21][22][23]. Interestingly, some natural products have also been proven to promote ICD by themselves, independently from the effects of antineoplastic drugs [24][25][26]. Therefore, currently, the combined delivery of chemotherapeutic drugs with natural bioactive products, in combination or not with immunotherapy, appears to be an exciting new area of research. ...
Phenolic compounds are bioactive phytochemicals showing a wide range of pharmacological activities, including anti-inflammatory, antioxidant, immunomodulatory, and anticancer effects. Moreover, they are associated with fewer side effects compared to most currently used antitumor drugs. Combinations of phenolic compounds with commonly used drugs have been largely studied as an approach aimed at enhancing the efficacy of anticancer drugs and reducing their deleterious systemic effects. In addition, some of these compounds are reported to reduce tumor cell drug resistance by modulating different signaling pathways. However, often, their application is limited due to their chemical instability, low water solubility, or scarce bioavailability. Nanoformulations, including polyphenols in combination or not with anticancer drugs, represent a suitable strategy to enhance their stability and bioavailability and, thus, improve their therapeutic activity. In recent years, the development of hyaluronic acid-based systems for specific drug delivery to cancer cells has represented a pursued therapeutic strategy. This is related to the fact that this natural polysaccharide binds to the CD44 receptor that is overexpressed in most solid cancers, thus allowing its efficient internalization in tumor cells. Moreover, it is characterized by high biodegradability, biocompatibility, and low toxicity. Here, we will focus on and critically analyze the results obtained in recent studies regarding the use of hyaluronic acid for the targeted delivery of bioactive phenolic compounds to cancer cells of different origins, alone or in combination with drugs.
... SK has also been proved to be an inducer of ROS in tumor cells, whereby inducing apoptosis and ICD [16]. Chen et al. has found that SK could induce ICD of melanoma B16 cells and enhance function of dendritic cells (DCs) [17]. Previous studies have con rmed that SK and BET inhibitor JQ1 can synergistically exert anti-tumor effects by reshaping the tumor immune microenvironment, but currently there is no research on the combination of SK and PD-1 monoclonal antibody [18]. ...
... CRT, HMGB1 and Hsp70 expression analysis. Tumor cell lysate samples were prepared as previously described [17]. The protein samples were resolved by SDS-PAGE using 10 or 15% stepwise gels. ...
... Chen et al. concluded that Shikonin could effectively increase the production of speci c DAMPs in B16 cells, including Hsp70, Hsp90, CRT and HMGB1. Shikonin-treated B16 cell lysates could induce DCs to a higher level of functional and phenotypic maturation, exhibiting high expression of CD86 and MHC class II and Th1 cells activation [17]. In our study, we also found that the percentage of CD11 + CD11b + DC and CD8 + T cells have been increased, suggesting that synergistic effect of SK with PD-1 blockade in CT26 xenograft mice model might due to DC activation and cytotoxic CD8 + T cells in ltration. ...
Background: PD-1 blockade has shown impressive clinical outcomes in colorectal cancers patients with high microsatellite instability (MSI-H). However, the majority of patients with colorectal cancer who present low microsatellite instability (MSI-L) or stable microsatellites (MSS) show little response to PD-1 blockade therapy. Here, we have demonstrated that Shikonin (SK) could induce cell death of CT26 cells via classically programmed and immunogenic pathways.
Methods and Results: SK promoted the membrane exposure of calreticulin and upregulated the expression of heat shock protein 70 (Hsp70). The upregulation of Hsp70 was dependent on ROS induced by SK and silencing of PKM2 in CT26 cells reverts ROS upregulation. Besides, SK synergizes with PD-1 blockade in CT26 xenograft mice model, with the increase of intramural DC cells and CD8⁺T cells. The expression of Hsp70 in tumor tissue was also increased in combinational SK plus αPD-1 therapy group.
Conclusions: Our study elucidated the potential role of ‘Shikonin-PKM2-ROS-Hsp70-ICD’ axis in the promotion of efficacy of PD-1 blockade in CRC treatments, providing a potential strategy and targets for improving the efficacy of PD-1 blockade in colorectal cancer.
... [52][53][54] In a study, increased DAMPs expression and a highly mature state of dendritic cells indicated SHK induced a significant ICD effect. 55 Natural killer cells, a type of innate lymphocytes, play a role in controlling cancer progression. A study confirmed SHK enhanced NK cell proliferation and cytotoxicity of Caco-2 cells by regulating the expression of P-ERK1/2 and P-AKt. ...
Malignant tumor, the leading cause of death worldwide, poses a serious threat to human health. For decades, natural product has been proven to be an essential source for novel anticancer drug discovery. Shikonin (SHK), a natural molecule separated from the root of Lithospermum erythrorhizon, shows great potential in anticancer therapy. However, its further clinical application is significantly restricted by poor bioavailability, adverse effects, and non-selective toxicity. With the development of nanotechnology, nano drug delivery systems have emerged as promising strategies to improve bioavailability and enhance the therapeutic efficacy of drugs. To overcome the shortcoming of SHK, various nano drug delivery systems such as liposomes, polymeric micelles, nanoparticles, nanogels, and nanoemulsions, were developed to achieve efficient delivery for enhanced antitumor effects. Herein, this review summarizes the anticancer pharmacological activities and pharmacokinetics of SHK. Additionally, the latest progress of SHK nanomedicines in cancer therapy is outlined, focusing on long circulation, tumor targeting ability, tumor microenvironment responsive drug release, and nanosystem-mediated combination therapy. Finally, the challenges and prospects of SHK nanomedicines in the future clinical application are spotlighted.
... PCD plays a signi cant function in immunity and is crucial for the coordination of the tumor microenvironment 31 . Several clinical trials have been conducted to explore the role of immunogenic PCD in cancer therapy [32][33][34] . ...
Background：Ovarian cancer has a high mortality rate, which has major contributing factors including advanced detection and recurrence susceptibility. For patients with ovarian cancer, optimal predictive models for therapy progression and medication sensitivity are currently lacking. Different types of programmed cell death (PCD) can predict prognosis and medication susceptibility in ovarian cancer and play a significant role in tumor growth.
Materials and methods: By analyzing 12 PCD patterns (apoptosis, necroptosis, ferroptosis, pyroptosis, netotic cell death, entotic cell death, lysosome-dependent cell death, parthanatos, autophagy-dependent cell death, oxeiptosis, and novel regulated cell death by alkaliptosis), Cox multifactorial regression analysis was used to screen a gene set of 20 genes that can each individually affect the prognosis of ovarian cancer. The gene set was then typed and modeled. The model was further validated and additional risk scoring was done. To learn more about the IC50 link between high and low-risk scores and chemotherapeutic drugs, the relationship between high and low-risk scores and the immune microenvironment was examined.
Results: The two examined subtypes of ovarian cancer have different key biological activities, according to the results of typing genes that can independently affect ovarian cancer survival, which combined with clinical characteristics to create a nomogram map with good predictive performance. Additionally, patients with ovarian cancer in the high-risk category were sensitive to Dasatinib, and the type was related to immune checkpoint genes and important tumor microenvironment components.
Conclusion: In short, by combining several cell death patterns, we created a new staging model that can precisely predict the clinical outcome and medication sensitivity of ovarian cancer patients.
... In real, SK has been demonstrated as provoke the production of various DAMPs in tumour cell lines, including CRT, HMGB1, GRPp78, HSP70, and HSP90. Other than that, tumour cells treated via SK and lipopolysaccharide (LPS) start DCs and can applied tumor-specific Th1 and Th17 CD4 T cells as distinct [36] . Further study revealed that the heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) is a particular protein aim of SK, and that its suppression is required for the production of the ICD-inducing signals. ...
Alternative bio actively chemicals may be found in natural goods and traditional herb medications, but only a few plant-information formulations have been rigorously studied and verified for their potential as medicinal therapies. The study of plant-derived elements' immunomodulation capabilities and their ability as provoke the immune system as combat various elemental disorders like cancer is, nonetheless, a promising area in current therapeutics information on plant-derived chemicals. This research showed how network pharmacology may be applied as define and validate natural individual elements or more complicated preparations as prospective cancer therapies information on their various aim capabilities in this research. We give a summary of the present state of understanding on network pharmacology, with a particular emphasis on various technical methods and their implications for cancer treatment.
... The latter, as a cancer vaccine system, often involves the effective employment of dendritic cells. Our laboratory in Academia Sinica, Taipei, performed a series of experimental studies [11][12][13][14], aiming to develop phytochemical-treated (PT) tumor cell lysate (TCL) and PT/TCL-pulsed (briefly co-cultured) and DC-based cancer vaccines for use against metastatic tumors. As seen in Figure 1, the experimental system makes good use of the iDC cultures. ...
... For instance, shikonin was shown to be highly effective as a key component of a cancer vaccine formulation for the treatment of tumor cell lysate (TCL)-pulsed, dendritic cell (DC)-based tumor vaccines [11]. Many of these activities were later shown to involve the DAMP signaling pathway [11][12][13] and/or the necroptotic autophagy activities [13]. ...
... For instance, shikonin was shown to be highly effective as a key component of a cancer vaccine formulation for the treatment of tumor cell lysate (TCL)-pulsed, dendritic cell (DC)-based tumor vaccines [11]. Many of these activities were later shown to involve the DAMP signaling pathway [11][12][13] and/or the necroptotic autophagy activities [13]. ...
Plenty of evidence has recently shown that various inflammatory activities at the local tissue, organ, or even the whole body (systemic) level are strongly linked to many life-threatening chronic diseases, most notably various cancers. However, only very limited information is available for making good use of our supporting immune-modulatory therapeutics for the treatment of cancers. This may result from a lack of studies on specific remedies for efficacious control or modulatory suppression of inflammation-related cancerous diseases. Our group and laboratories were fortunate to have initiated and consistently pursued an integrated team-work program project, aimed at investigating selected medicinal herbs and the derived, purified phytochemical compounds. We focused on the study of key and specific immune-signaling mechanisms at the cellular and molecular levels. We were fortunate to obtain a series of fruitful research results. We believe that our key findings reported herein may be helpful for proposing future thematic and integrated research projects that aim to develop future phytochemical drugs against cancers. The mechanisms of the cellular and molecular systems involved in inflammation are becoming increasingly recognized as keystones for the development of future therapeutic approaches for many chronic and cancerous diseases. Recently, the immune checkpoint inhibitors such as antibodies against PD-1 and/or PD-L1 have been shown to be too expensive for general clinical use, and their effects far from optimal, often showing little or no effect or only short-term efficacy. These results point to the need for developing future immune-regulatory or modulatory therapeutics.
... It has been shown that BTZ enhances the DC-mediated T cell response via promoting the exposure of HSP90 on the cell membrane of dying myeloma cells [63]. Shikonin is a phytochemical compound that can either trigger the extrinsic apoptotic pathway by stimulating death receptors followed by activation of caspase-8, or the intrinsic apoptotic pathway by activating the apoptosis regulator Bcl-2-like protein 4 (BAX), leading to cytochrome c release and activation of caspase-9 [64]. B16 mouse melanoma cells, cultured in the presence of shikonin for 24 h, exhibited elevated expression of HSP70, HSP90, CRT, and HMGB1 [55]. ...
The new and increasingly studied concept of immunogenic cell death (ICD) revealed a previously unknown perspective of the various regulated cell death (RCD) modalities, elucidating their immunogenic properties and rendering obsolete the notion that immune stimulation is solely the outcome of necrosis. A distinct characteristic of ICD is the release of danger-associated molecular patterns (DAMPs) by dying and/or dead cells. Thus, several members of the DAMP family, such as the well-characterized heat shock proteins (HSPs) HSP70 and HSP90, the high-mobility group box 1 protein and calreticulin, and the thymic polypeptide prothymosin α (proTα) and its immunoreactive fragment proTα(100–109), are being studied as potential diagnostic tools and/or possible therapeutic agents. Here, we present the basic aspects and mechanisms of both ICD and other immunogenic RCD forms; denote the role of DAMPs in ICD; and further exploit the relevance of human proTα and proTα(100–109) in ICD, highlighting their possible clinical applications. Furthermore, we present the preliminary results of our in vitro studies, which show a direct correlation between the concentration of proTα/proTα(100–109) and the levels of cancer cell apoptosis, induced by anticancer agents and γ-radiation.
