Characterization of cyclotide Vdif A from Viola diffusa inhibits non-small cell lung cancer cells via regulation of CKS2.

Cancer cachexia (CC) is a highly debilitating syndrome characterized by loss of body and muscle weight affecting most advanced cancer patients. The receptor for advanced glycation end-products (RAGE) is expressed by several cell types and sustains the inflammatory response in acute and chronic diseases. Total ablation of RAGE (Ager-/- mice) translates into restrained CC and increased survival in tumour-bearing mice. RAGE, which is not expressed in adult healthy myofibres, is re-expressed in atrophying myofibres in cancer conditions. However, the specific contribution of muscular RAGE to CC was unknown. Using an HSA/Cre-loxP system, we generated a tamoxifen-inducible conditional AgermKO mouse model in which RAGE is selectively ablated in myofibres. Tamoxifen-treated AgermKO, Agerflox and Ager-/- mice were subcutaneously injected with Lewis lung carcinoma (LLC) cells, and body changes and survival were monitored until 25 dpi, when histological, molecular and proteomic analyses were performed in tumour-bearing and control mice. Muscle samples of pre-cachectic and cachectic pancreatic cancer patients were analysed to validate the results. Compared with LLC-Agerflox mice, LLC-AgermKO mice showed reduced (7.5% [p = 0.004] vs. 15.1% [p < 0.0001]) body weight loss, no significant reduction of hind-limb muscle mass and strength and myofibre cross-sectional areas, increased survival (69.2% vs. 42.9% mice alive at 25 dpi) and restrained muscle and serum pro-inflammatory factors. Mechanistically, AgermKO muscles resist cancer-induced atrophy by maintaining an active Akt-GSK-3β-PGC-1α pathway, and increasing the synthesis of myosin heavy chain (MyHC)-I and -IIa (71.8% [p = 0.008] and 73.9% [p = 0.002] increase, respectively) along with a 76.3% (p = 0.008) increase in hybrid MyHC-I/IIa myofibres. Distinct proteomic signatures characterize muscles of tumour-bearing mice in dependence on RAGE expression, supporting a protective effect of RAGE ablation in muscles. LLC/AgermKO muscles showed increased amounts of several enzymes involved in glycolysis and glucose catabolism, typical of Warburg metabolism. Noteworthy, muscles of pre-cachectic and cachectic cancer patients showed ~3-fold increase (p < 0.05) in RAGE amounts and reduced Akt-GSK-3β-PGC-1α pathway, compared with healthy control subjects. Our data provide evidence that RAGE engagement at myofibre level drives loss of body and muscle weights and inflammation in cancer conditions. RAGE ablation in muscles confers resistance to CC through myofibre remodeling and glycolytic reprogramming. On the clinical side, the overexpression of RAGE is an early event in muscles of cancer patients, suggesting a role for RAGE in the onset of the cachectic syndrome. Thus, the molecular targeting of RAGE might be useful to counteract cachexia and prolong survival in cancer patients.

Design, synthesis, and biological evaluation of a hydrophilic 20-O-glycyl ester prodrug of 10-methoxycamptothecin against lung cancer.

Improving the bioavailability of nintedanib by formulating inhalable ufasomes as a targeted therapy for non-small cell lung cancer.

Deziyangxin (DZYX) is a classical Tibetan multi-herb prescription recorded in Jing Zhu Ben Cao and traditionally used for conditions characterized by blood stasis, phlegm-heat accumulation, and tumor-like masses. Despite its long-standing clinical use, the pharmacological basis underlying its potential anti-tumor activity remains insufficiently understood. This study aimed to evaluate the anti-tumor efficacy of DZYX in non-small cell lung cancer (NSCLC) and to explore its potential mechanisms with emphasis on tumor immune microenvironment modulation. Serum pharmacochemistry was performed using LC-MS/MS to identify circulating constituents of DZYX. A syngeneic Lewis lung carcinoma (LLC) mouse model was established in mice, which were treated with DZYX by oral gavage at 78.5mg/kg or 157mg/kg for 25 days. Tumor growth and histopathology were evaluated, while transcriptomic analysis, network pharmacology, flow cytometry, and immunohistochemistry were used to investigate molecular pathways and immune cell infiltration. LC-MS/MS identified 167 circulating components derived from DZYX. DZYX treatment significantly suppressed tumor growth and induced tumor necrosis in LLC-bearing mice. Transcriptomic and enrichment analyses indicated modulation of chemokine signaling, T-cell activation, and innate immune pathways. Flow cytometry and immunohistochemistry further demonstrated increased infiltration of CD4+ and CD8+ T cells, NK cells, and B cells within tumor tissues and spleens. Enhanced intratumoral Granzyme B and IFN-γ signals suggested increased cytotoxic immune activity. DZYX exerts anti-tumor activity in NSCLC associated with remodeling of the tumor immune microenvironment and multi-pathway molecular regulation. These findings provide mechanistic support for the traditional use of DZYX in tumor-related disorders and highlight its potential as an immunomodulatory candidate for lung cancer therapy.

XIAOSHUI formula inhibits malignant pleural effusion by targeting the STC1/p65/CXCL5 axis to reprogram tumor-associated macrophages.

We aimed to investigate the utility of Au-incorporated hyaluronic acid nanoparticles (Au/HA NPs) for improving the therapeutic efficacy of ablative radiotherapy (RT) for tumor control and microenvironment remodeling. HA-functionalized NPs exhibited uniform size, stability, and efficient SN38 encapsulation. Au incorporation increased NP diameter and reduced surface charge while remaining stable. HA and Au/HA NPs were efficiently internalized by lung cancer cells, with free HA pretreatment suppressing internalization. Moreover, Au/HA NP internalization strongly downregulated CD44 expression in lung cancer cells, confirming CD44-mediated internalization. In vitro, Au/HA NPs enhanced radiation-induced G2/M phase arrest and γH2AX foci formation with increased DNA double-strand breaks. Au/HA NPs and RT induced immunogenic cell death (ICD) in lung cancer cells, characterized by elevated reactive oxygen species, increased calreticulin surface expression, and extracellular adenosine triphosphate release. Tumor control, survival, immune infiltration, and systemic effects were investigated in vivo using A549 xenografts and Lewis lung carcinoma synchronous flank-lung tumor models. Au/HA NPs and ablative RT decreased tumor growth, reduced lung tumor burden in non-irradiated areas, and prolonged survival. This therapeutic combination led to increased infiltration of natural killer (NK), NK T, CD8+ T, and dendritic cells and decreased regulatory T cells, suggesting robust immunological activation. Biodistribution studies confirmed CD44-targeted tumor-specific NP accumulation. No substantial toxicity was observed. In conclusion, Au/HA NPs and ablative RT induced ICD in vivo. Au/HA NPs enhanced local and systemic immunity via radiosensitization and ICD. This NP-assisted approach may improve RT efficacy in lung cancer.

Ablation of ASF1B mitigates the proliferation of A549 cells and enhances anti-PD-L1 therapy by regulating ferroptosis.

FcRn function alone cannot account for elevated IgG clearance in the Lewis lung carcinoma model of cancer cachexia.

Moringa Oleifera (MO) is a potential plant-derived cancer treatment. Its anticancer effects are multi-targeted and include immunological stimulation, regulation of oxidative stress, and induction of apoptosis. To fully evaluate the implications of MO on lung cancer in in vitro and in vivo models, this mini-review focuses on linking these observations with cytotoxic, molecular, and immunological effects. Using PubMed, Scopus, and Web of Science databases, comparative studies on MO extracts, phytochemicals, and nanoparticle-delivered anticancer activities against lung cancer were identified. Fourteen studies were included: twelve in vitro (primarily on A549 NSCLC adenocarcinoma cells) and two in vivo. Measures including immune responses, ROS modulation, apoptosis markers, and cell viability were used to evaluate the results. In vitro models, MO consistently reduced the viability of lung cancer cells, induced apoptosis by the mitochondrial pathway (Bax/Bcl-2 switch, cytochrome c release, caspase activation), and blocked oncogenic signaling pathways (NF-kB, EGFR, STAT3, VEGF). Nanoparticle formulations (AuNPs, AgNPs, CuO NPs, PdNPs) demonstrated greater cytotoxicity and pro-apoptotic activity than crude preparations. In vivo, MO leaf extracts suppressed tumor growth in urethane-treated rats in a manner comparable to cisplatin, and MO polysaccharides repolarized tumor-associated macrophages from an M2 to an M1 phenotype and enhanced CD4⁺/CD8⁺ T-cell infiltration in Lewis lung carcinoma mice. This mini-review highlights the potent anticancer activity of Moringa oleifera against lung cancer through apoptosis induction, oxidative stress modulation, and inhibition of oncogenic signaling and angiogenesis. Despite promising in vitro and in vivo results, further standardized and clinical studies are required to validate its therapeutic efficacy and safety. Through a variety of mechanisms, including apoptosis, redox balance, angiogenesis, and immune modulation, this review demonstrated Moringa oleifera's potent anticancer activity in both in vivo and in vitro models of lung cancer. These findings suggest that Moringa oleifera represents a promising preclinical candidate for further investigation as a supportive strategy in non-small cell lung cancer, pending validation through standardized in vivo and clinical studies.

Worldwide, lung cancer is the most common cause of cancer-related deaths. Molecular targeted therapies and immunotherapies for non-small-cell lung cancer (NSCLC) have improved outcomes markedly over the past two decades. However, the vast majority of advanced NSCLCs become resistant to current treatments and eventually progress. A traditional Chinese medicine (TCM) formula of Shuangshen granules (SSG) has demonstrated potential in alleviating cancer side effects and improving survival rate. Despite clinical evidence supporting its benefit, there is still insufficient understanding of the active compounds in SSG and their underlying mechanisms, which limits its broader clinical application. Lewis lung carcinoma (LLC) tumor-bearing mouse model was established to assess the efficacy of combined SSG and anti-PD-1 therapy in vivo, and myeloid-derived suppressor cells (MDSC) and CD8+T cells were isolated for in vitro co-culture experiments, while pathological examination was conducted using hematoxylin and eosin (HE). The expression of PD-1, TIM-3, CTLA-4, LAG-3, Arg-1, IDO, iNOS, PD-L1 and Gal-9 was detected using immunohistochemistry (IHC), immunofluorescence, and flow cytometry and Western blotting. The expression of IL-2, TNF-α and IFN-γ were detected by reverse transcription-quantitative polymerase chain reaction (qPCR). Concentrations of IL-10 and TGF-β were measured by enzyme-linked immunosorbent assay (ELISA). Network pharmacology and molecular docking were utilized to screen for potential therapeutic targets and intervening signaling pathways of SSG in lung adenocarcinoma (LUAD). The predictions derived from this approach were further verified using Western blotting. In vivo experiments using LLC xenograft mice demonstrated that SSG suppressed tumor growth in a dose-dependent manner, with high-dose SSG showing optimal efficacy in inhibiting tumor angiogenesis and cell proliferation. SSG enhances anti-tumor immunity by reducing T cell exhaustion and MDSC-mediated immunosuppression, with SSG + anti-PD-1 combination therapy synergistically optimizing the tumor immune microenvironment. Network pharmacology analysis revealed 5 hub targets (IL2, STAT3, HSP90AA1, LGALS3, and FGF2) associated with immune, LUAD, and active ingredients of SSG, with significant enrichment in the PI3K-Akt pathway. Compared with the control group, the protein expression levels of p-PI3K and p-Akt in the SSG group were significantly down-regulated, indicating that the PI3K-Akt pathway may be inhibited. SSG could dose-dependently inhibit LLC tumor growth in mice and exert antitumor effects by alleviating T-cell exhaustion and MDSC-mediated immunosuppression. Notably, IL2, STAT3, HSP90AA1, LGALS3 and FGF2, as potential targets of SSG, were significantly enriched in the PI3K-Akt pathway, which provide a novel perspective for the treatment of LUAD.

Circulating low-density neutrophils as biomarkers of resistance to first-line anti-PD-1/PD-L1 immunotherapy in non-small cell lung cancer.

Eliciting calreticulin (CRT) surface exposure is essential for triggering immunogenic cell death (ICD). However, stanniocalcin 1 (STC1) suppresses CRT translocation by sequestering it within mitochondria, limiting ICD induction in tumours. Here, we show that silencing STC1 enhances CRT surface exposure in Lewis lung carcinoma (LLC) cells when combined with paclitaxel (PTX), converting dying tumour cells into an in-situ vaccine that drives immunoprevention of tumour growth. To maximize this therapeutic synergy, we engineered a nanoplatform co-delivering siSTC1 and PTX, in which PTX is covalently conjugated to a sphingolipid and siSTC1 is electrostatically encapsulated (siSTC1/LNP-PTX). This system improves pharmacokinetics, synchronizes co-delivery to tumours, and enhances intratumoral exposure. Consequently, it amplifies CRT expression, promotes antigen-presenting cell-mediated phagocytosis and antigen presentation, and elicits robust cytotoxic T cell responses in LLC models. Moreover, siSTC1/LNP-PTX sensitizes tumours to PD-1 blockade. Our nanosystem, which unlocks ICD potential by silencing STC1, represents a paradigm-shifting approach to cancer immunotherapy.

Cancer cachexia is a debilitating syndrome characterized by severe skeletal muscle wasting, which significantly impairs patient quality of life and survival. Indoleamine 2,3-dioxygenase 1 (IDO1), a key enzyme in tryptophan (Trp) metabolism, is often upregulated in cancers, but its specific role in driving lung cancer-associated cachexia remains inadequately defined. This study investigated the mechanistic role of Ido1 in cancer cachexia and evaluated the therapeutic potential of its inhibition. We established Lewis lung carcinoma (LLC) models in C57BL/6 mice using wild-type, Ido1-overexpressing (Ido1-OE) and Ido1-knockout (Ido1-KO) cells. Muscle mass, tumour growth and metabolic changes were assessed invivo. Transcriptomic and targeted metabolomic analyses were performed on muscle and serum samples. Invitro, we examined the effects of tumour-conditioned media, the Trp metabolite kynurenine (Kyn) and Trp supplementation on C2C12 myotube atrophy. Invivo experiments verified the efficacy of the Ido1 inhibitor palmatine hydrochloride (PAL). Molecular pathways were analysed via western blot and qPCR. Compared to LLC mouse models, Ido1-OE significantly exacerbated tumour growth and cachexia, leading to a significant decrease in lean body weight, gastrocnemius and tibialis anterior muscle weights (p < 0.01, p < 0.0001, p < 0.001). Gastrocnemius muscle fibre cross-sectional area significantly decreased in the Ido1-OE group (p < 0.0001). Transcriptomic analysis revealed that Ido1-OE activated pro-inflammatory and protein degradation pathways (upregulating MuRF1/Atrogin1, p < 0.05) while suppressing anabolic signalling pathways (oestrogen pathways, p < 0.01). Metabolomics analysis revealed unique metabolic signatures in Ido1-OE mice: Trp depletion and Kyn accumulation. Invitro experiments demonstrated that Ido1-OE enhanced LLC cell proliferation and migration capabilities (p < 0.0001, p < 0.0001). Tumour-conditioned medium (TCM) derived from Ido1-OE tumours significantly induced C2C12 myotube atrophy (p < 0.01). Similarly, direct treatment with Kyn led to dose-dependent muscle fibre shrinkage, with significant atrophy observed at 30 μM (p < 0.01) and 100 μM (p < 0.0001). Notably, the myotube atrophy induced by Kyn was significantly reversed by the addition of supplemental Trp (p < 0.0001). Compared with the Ido1-OE group, PAL treatment reduced gastrocnemius and tibialis anterior atrophy (p < 0.01; p < 0.05). Mechanistically, PAL inhibited the mRNA expression levels of MuRF1/Atrogin1 (p < 0.0001, p < 0.001), as well as their corresponding protein levels (p < 0.0001, p < 0.0001). Furthermore, PAL restored the phosphorylation level of mTOR (p < 0.001), as well as the mRNA expression of myosin heavy chain (p < 0.01). Our findings demonstrate that Ido1 accelerates muscle atrophy and cancer cachexia by driving a metabolic reprogramming centred on the Trp-Kyn pathway. Pharmacological inhibition of Ido1 with PAL effectively mitigates these effects, positioning Ido1 as a promising therapeutic target for treating cancer cachexia.

Fangji Huangqi Decoction (FJHQ), a traditional Chinese medicine formula derived from "Jin Gui Yao Lue," has been reported to exhibit immunomodulatory and anti-inflammatory effects. This study investigates its potential anti-tumor mechanism via the enhancement of CD8+ T cell-mediated immunity. In murine B16 melanoma and Lewis lung carcinoma models, FJHQ significantly suppressed tumor growth and enhanced CD8+ T cell infiltration and cytotoxicity. CD8+ T cell depletion abrogated these effects, confirming their necessity. Invitro, FJHQ treatment augmented antigen presentation by tumor cells through upregulation of MHC-I expression, leading to improved CD8+ T cell recognition and killing. Mechanistically, FJHQ inhibited autophagic flux in tumor cells, as evidenced by mCherry-GFP-LC3 reporter assays and electron microscopy, resulting in impaired lysosomal function and elevated MHC-I levels. Furthermore, combining FJHQ with anti-PD-1 therapy synergistically promoted CD8+ T cell activity, increased granzyme B and IFN-γ expression, and induced substantial tumor regression. These findings demonstrate that FJHQ enhances antitumor immunity by suppressing autophagy and elevating MHC-I expression, thereby sensitizing tumors to PD-1 blockade. FJHQ represents a promising immunoadjuvant for cancer therapy.

Cancer cachexia is a multifactorial metabolic syndrome that profoundly reduces muscle mass, strength, efficacy of chemotherapy, and survival, yet no effective therapy exists. Unacylated ghrelin (UnAG), the predominant form of circulating ghrelin, promotes muscle growth and mitochondrial bioenergetics, but its role in cancer cachexia remains unknown. Four- to 5-mo-old male C57Bl/6N mice were assigned to three groups: nontumor-bearing (NTB), tumor-bearing (TB), and tumor-bearing treated with UnAG (TB + UnAG). Lewis lung carcinoma cells were inoculated subcutaneously in the flank of the mice. Body weight, food intake, and tumor size were monitored for 4 wk. Lower limb muscle mass, contractile function, mitochondrial respiration, and reactive oxygen species (ROS) production were measured, in conjunction with Western blot, proteomic, and immunohistochemical analyses. Compared with NTB controls, TB mice exhibited marked loss of muscle mass and function, whereas UnAG treatment preserved ∼50% of the muscle mass and ∼70% of the contractile force. UnAG enhanced mitochondrial oxygen consumption, reduced ROS generation, and preserved mitochondrial DNA copy number and downregulated DNA mutation frequency. TB mice demonstrated increased oxidative stress and activation of protein degradation pathways, along with neuromuscular junction disruption-both of which were normalized by UnAG. These findings collectively demonstrate that UnAG mitigates cancer cachexia by modulating mitochondrial bioenergetics, oxidative and proteolytic stress, and neuromuscular junction integrity. UnAG represents a promising therapeutic candidate that may mitigate cachexia and improve both chemotherapy efficacy and the quality of life of patients with cancer.NEW & NOTEWORTHY Unacylated ghrelin prevents muscle wasting and maintains neuromuscular junction integrity and contractile function in cancer cachexia. It enhances mitochondrial respiratory capacity through elevated mitochondrial DNA copy number while limiting DNA mutation. It reduces mitochondrial reactive oxygen species (mtROS) generation, oxidative stress, and proteasome-mediated proteolysis-driven muscle degradation.

The maintenance of skeletal muscle is of pivotal importance, as its loss is often associated with progressive pathologies, generally worsening the prognosis. Increased levels of vitamin D binding protein (VDBP) were reported in diseases susceptible to muscle wasting, including several tumors. We hypothesized that VDBP might participate in muscle wasting and investigated its direct effects on skeletal muscle homeostasis. Here, we demonstrate that VDBP induces atrophy independently of vitamin D. In C2C12 myotubes, we identified intracellular actin dynamics perturbation and subsequent mitochondrial fragmentation as the main molecular mechanisms of VDBP-induced atrophy. Coherently, the ectopic introduction of VDBP in mice lacking the protein (Gc-knockout mice) induced muscle atrophy and decreased strength. Finally, we present proof-of-concept evidence that VDBP contributes to cancer-associated muscle wasting in Lewis lung carcinoma (LLC)-bearing male mice. Altogether, these findings provide novel insights into the biological function of VDBP as a pro-atrophic hormone with potential implications for the treatment of muscle wasting.

Drug-induced lung disease (DILD) is a severe adverse event of cancer treatment. Several clinical reports have demonstrated an association between DILD and tumor progression. However, the underlying mechanism remains unclear. This study aimed to elucidate the role of tumor-bearing status in the development of DILD. We prepared a subcutaneous Lewis lung carcinoma (LLC) and KLN205-bearing model. To trigger DILD, bleomycin (BLM) was administered subcutaneously. mRNA expression associated with endothelial activation (PAI-1, vWF, and ICAM-1), inflammatory cell infiltration, and alveolar wall thickness was assessed by using bronchioalveolar lavage fluid (BALF) and lung tissue. Additionally, the role of high-mobility group box1 (HMGB1) in tumor-bearing status was examined. Compared with control mice, LLC- and KLN205-bearing mice showed a tendency toward increased expression of at least one of PAI-1, vWF, and ICAM-1 on endothelium, along with inflammatory cell infiltration in the lungs. BLM-treated mice with LLC showed more inflammatory cell infiltration than BLM-treated mice, accompanied by a significant increase in PAI-1, vWF, and ICAM-1 expression on endothelium. Moreover, BLM-treated mice with LLC exhibited pronounced alveolar wall thickening. In LLC-bearing mice, serum HMGB1 levels were significantly higher compared with control mice. Additionally, inflammatory cell infiltration in the lungs tended to be increased by the intraperitoneal injection of HMGB1, which was accompanied by increased expression of vWF and ICAM-1 on endothelium. This study showed that tumor-bearing status elicits proinflammatory activation in endothelial cells and inflammatory cell infiltration into the lungs that aggravates DILD caused by BLM.

Glucocorticoids are frequently administered to alleviate therapy-related side effects in cancer patients, yet their role in tumor progression remains controversial and mechanistically unresolved. Here, we demonstrate that the long-acting glucocorticoid dexamethasone (Dex) exerts antitumor effects that are mediated by neutrophils. In murine models of Lewis lung carcinoma (LLC) and B16F10 melanoma, Dex markedly suppressed tumor growth and prolonged survival of tumor-bearing mice. These effects were independent of adaptive immunity, macrophages, and tumor cell-intrinsic glucocorticoid signaling, but required functional glucocorticoid receptor (GR) signaling in neutrophils. Dex-treated neutrophils exhibited longer survival and higher cytotoxicity toward tumor cells via increased production of reactive oxygen species (ROS). Disruption of this GR-ROS axis, either through neutrophil-specific GR deletion or pharmacological inhibition of ROS, abolished the antitumor activity of Dex. Together, these findings uncover a neutrophil-mediated tumoricidal function of Dex and suggest that neutrophil GR-ROS signaling may be harnessed for cancer therapy.

To investigate the anti-tumor effects and immuno-inflammatory-metabolic mechanisms of Buzhong Yiqi Granules (BYG) in alleviating cancer-related fatigue (CRF) in lung cancer. UPLC-Q-TOF/MS detected compounds in BYG. CRF was induced in mice via Lewis lung carcinoma cell inoculation, with daily BYG treatment for 14days. Anti-fatigue efficacy was evaluated through behavioral tests and ATP content detection. Anti-tumor effects were assessed via histopathology, tumor volume, and weight. Immuno-inflammatory-metabolic mechanisms were evaluated using inflammatory cytokines (IL-6, TNF-α, IFN-γ) and immune cell subsets (CD4+, CD8+ T cells) in serum and tumor tissues. Ninety-four active compounds were identified in BYG. In the CRF mouse model, BYG prolonged swimming time, reduced tail suspension immobility, increased muscle ATP to exert anti-fatigue effects; it inhibited tumor growth and metastasis by reducing volume and downregulating EGFR/CD34/MMP-9; it also modulated inflammation and immunity by lowering IL-6/TNF-α, elevating IFN-γ, enhancing CD4 + T cells and restoring the CD4 + /CD8 + ratio. BYG alleviates CRF via multi-target mechanisms involving energy metabolism, tumor inhibition, and immune-inflammatory regulation, supporting its clinical use in CRF and potential as lung cancer adjuvant therapy.