Toxicology and Applied Pharmacology

Caulerpin suppresses tumor-associated angiogenesis and tumor growth via Hippo signaling in cervical cancer

Caulerpin, a bisindole alkaloid derived from green algae of the genus Caulerpa, has exhibited a promising anti-proliferative effect on various tumor cells in vitro. However, its pharmacological potential has not been intensively explored in cervical cancer. In this study, the antitumor property of caulerpin was assessed in cervical cancer cells (HeLa and SiHa cells) and xenograft mouse models. It was found that caulerpin significantly inhibited cell growth, colony formation, migration, invasion and induced cell apoptosis in HeLa and SiHa cells, and inhibited the tumor growth of HeLa xenografts in vivo. In human umbilical vein endothelial cells (HUVECs), caulerpin significantly suppressed proliferation, migration, and tube formation with (indirect) or without (direct effects) tumor-conditioned medium stimulation. Mechanically, caulerpin treatment blocked angiogenesis in vitro and in vivo through suppressing YAP1 nuclear translocation and angiogenesis-related proteins expression, including HIF-1α and VEGFA expression in tumor cells, VEGFR2 and CD31 expression in HUVECs. TDI-011536 (a specific inhibitor of Hippo) effectively reversed the effect of caulerpin. These results demonstrate the anti-angiogenic effect of caulerpin in cervical cancer that Hippo/YAP1 mediates tumor-endothelial cell interaction via VEGF/VEGFR2 signaling, which supports it as a natural product for the management of cervical cancer.

EP300 promotes cervical cancer progression through a Wnt/β-catenin–dependent HIF-1α signaling axis

This study aimed to investigate the role of EP300 in cervical cancer progression, focusing on its functional effects related to pyroptosis under Wnt/β-catenin-dependent conditions. In SiHa and HeLa cells, EP300 was silenced using lentiviral-mediated knockdown under hypoxic conditions, and subsequent changes in cell proliferation, inflammatory cytokine secretion, and pyroptosis-related protein expression were evaluated. A subcutaneous HeLa xenograft model was established to examine the in vivo effects. Hypoxia-inducible factor 1-alpha (HIF-1α) expression was also analyzed. The results showed that hypoxia significantly increased the expression levels of EP300, β-catenin, and HIF-1α. EP300 knockdown was associated with decreased Wnt/β-catenin signaling and, under Wnt/β-catenin-dependent conditions, resulted in reduced HIF-1α expression and enhanced pyroptosis-related phenotypes. Activation of Wnt/β-catenin signaling by HLY78 partially reversed these alterations. In vivo, silencing EP300 suppressed tumor growth, decreased Ki67 expression, and increased pyroptosis markers, while activation of the Wnt/β-catenin pathway partially restored proliferation and reduced pyroptosis. Taken together, loss of EP300 function impedes cervical cancer progression by affecting the Wnt/β-catenin signaling axis and inducing concomitant changes in HIF-1α expression. This study provides functional evidence supporting the role of EP300 in the progression of cervical cancer under hypoxic conditions.

The anti-cancer mechanism of Celastrol by targeting JAK2/STAT3 signaling pathway in gastric and ovarian cancer

Celastrol is a natural triterpene exhibiting significant and extensive antitumor activity in a wide range of cancer. Due to unfavorable toxicity profile and undefined mechanism, Celastrol's application in clinical cancer therapy remains limited. Herein, we elucidate the pharmacological mechanism of Celastrol's anticancer effects, with a focus on STAT3 signaling pathway in cancers with high incidence of metastasis. The safety profile of Celastrol were assessed in mice. In vitro analysis was performed in gastric cancer and ovarian cancer to assess the cytotoxicity, induction of reactive oxygen species (ROS) of Celastrol using STAT3 knockout cancer cells. Effects of Celastrol on STAT3 activation and transcription activity, JAK2/STAT3 signaling protein expression were assessed. Additionally, proteomic contrastive analysis was performed to explore the molecular association of Celastrol with STAT3 deletion in cancer cells. Celastrol has no obvious toxic effect at 1.5 mg/kg/day in a 15 days' administration. Celastrol inhibits tumor growth and increases ROS in a STAT3 dependent manner in gastric and ovarian cancer celllines. On molecular level, it downregulates IL-6 level and inhibits the JAK2/STAT3 signaling pathway by suppressing STAT3' activation and transcription activity. Proteomic contrastive analysis suggests a similar cellular mechanism of action between Celastrol and STAT3 deletion on regulating cancer progression pathways related to migration and invasion. Our research elucidates the anti-cancer mechanism of Celastrol through targeting the JAK2/STAT3 signaling pathway in cancer with high incidence of metastasis. This study provides a solid theoretical basis for the application of Celastrol in cancer therapy.

Phellopterin attenuates ovarian cancer proliferation and chemoresistance by inhibiting the PU.1/CLEC5A/PI3K-AKT feedback loop

Ovarian cancer is known as the second leading cause of gynecologic cancer-associated deaths in women worldwide. Developing new and effective compounds to alleviate chemoresistance is an urgent priority in ovarian cancer. Here, we aimed to reveal the biological function and underlying mechanisms of phellopterin, a naturally sourced ingredient of Angelica dahurica, in ovarian cancer progression as well as evaluate the therapeutic potential of phellopterin in ovarian cancer patients. In this investigation, we found that phellopterin mitigated DNA replication and induced cell cycle arrest, apoptosis, and DNA damage, attenuating cell proliferation and chemoresistance of ovarian cancer. Interestingly, bioinformatics analyses of data from our RNA sequencing and The Cancer Genome Atlas ovarian cancer dataset suggested that phellopterin presented anti-cancer activities in ovarian cancer cells by modulating signals affecting ovarian cancer progression and identified phellopterin as a potential compound in improving ovarian cancer patients' prognosis. In addition, the C-Type Lectin Domain Containing 5A (CLEC5A) was demonstrated as a downstream effector of phellopterin and involved in a positive PU.1/CLEC5A/PI3K-AKT feedback loop. Interestingly, phellopterin might inactivate the positive feedback circuit to suppress ovarian cancer progression. Collectively, our investigation revealed that phellopterin mitigated ovarian cancer proliferation and chemoresistance through suppressing the PU.1/CLEC5A/PI3K-AKT feedback loop, and predicted phellopterin as a new and effective cytotoxic drug and CLEC5A as a potential target for the treatment of ovarian cancer.

Association of urinary arsenic, polycyclic aromatic hydrocarbons, and metals with cancers among the female population in the US

Cancers that primarily affect women in the US include breast, uterine, and cervical cancers. There may be associations between these different types of cancer in women and environmental pollutant exposure. This study aimed to assess seven species of arsenic, six polycyclic aromatic hydrocarbon (PAH) compounds, and fourteen different metal concentrations in urine and their correlation with cancer among women. We conducted a cross-sectional analysis using 2011--2012 to 2015-2016 National Health and Nutrition Examination Survey data (n = 4,956) and logistic regression modeling of the complex weighted survey design. Breast cancer was inversely correlated with arsenocholine (3rd quantile), monomethylarsonic acid (4th quantile), manganese (4th quantile), and antimony (3rd, 4th quantiles). Cervical cancer was inversely correlated with 3-hydroxyfluorene (3rd quantile), molybdenum (2nd, 4th quantiles), antimony (3rd quantile), tin (4th quantile), and thallium (4th quantile) exposure and positively associated with arsenic acid (3rd quantile), arsenobetaine (2nd, 4th quantiles). Uterine cancer was correlated with 1-hydroxynaphthalene (3rd, 4th quantiles), 2-hydroxynaphthalene (4th quantile), 1-hydroxyphenathrene (2nd, 4th quantiles), 1-hydroxypyrene (3rd quantile), cobalt (2nd, 3rd quantiles) and inversely with mercury (4th quantile). This study determined breast cancer and arsenic and some metal species exposure, indicating an inverse association. Arsenic acid and arsenobetaine exposure showed a positive correlation with cervical cancer. For uterine cancer, the correlations for the PAH compounds and cobalt showed a positive correlation, and the arsenic species and mercury were inversely associated. Further research is required to establish or refute the findings.

A natural product, voacamine, sensitizes paclitaxel-resistant human ovarian cancer cells

Most women with ovarian cancer are treated with chemotherapy before or after surgery. Unfortunately, chemotherapy treatment can cause negative side effects and the onset of multidrug resistance (MDR). The aim of this study is to evaluate the chemosensitizing effect of a natural compound, voacamine (VOA), in ovarian (A2780 DX) and colon (LoVo DX) cancer drug-resistant cell lines which overexpress P-glycoprotein (P-gp), in combination with paclitaxel (PTX), or doxorubicin (DOX) or 5-fluorouracil (5-FU). VOA, a bisindole alkaloid extracted from Peschiera fuchsiaefolia, has already been shown to be effective in enhancing the effect of doxorubicin, because it interferes with the P-gp function. Ovarian cancer cytotoxicity test shows that single treatments with VOA, DOX and PTX do not modify cell viability, while pretreatment with VOA, and then PTX or DOX for 72 h, induces a decrease. In colon cancer, since 5-FU is not a-substrate for P-gp, VOA has no sensitizing effect while in VOA + DOX there is a decrease in viability. Annexin V/PI test, cell cycle analysis, activation of cleaved PARP1 confirm that VOA plus PTX induce apoptotic cell death. Confocal microscopy observations show the different localization of NF-kB after treatment with VOA + PTX, confirming the inhibition of nuclear translocation induced by VOA pretreatment. Our data show the specific effect of VOA which only works on drugs known to be substrates of P-gp.

Ivermectin-induced cell death of cervical cancer cells in vitro a consequence of precipitate formation in culture media

The anthelmintic ivermectin has been reported to possess anticancer and antiviral efficacy. However, the effective concentrations reported in vitro are near the predicted aqueous solubility limit for this hydrophobic drug. We observed that ivermectin-induced cell death in two cervical cancer cell lines correlated with the formation of solid ivermectin aggregates in both serum-free and serum-supplemented culture media. Filtration of ivermectin particles >0.2 μm abolished these cytolytic effects in both cell lines. An inhibitory effect on cell proliferation persisted for filtered solutions, but only for ivermectin concentrations higher than reported to be clinically attainable in humans. In addition to the importance of distinguishing between free and bound drug in solution, our data emphasize the importance of acknowledging the likely solubility limit of hydrophobic drugs when assessing their in vitro cytotoxicity.

Effect of substituents at the C3´, C3´N, C10 and C2-meta-benzoate positions of taxane derivatives on their activity against resistant cancer cells

We tested the effect of substituents at the (1) C3´, C3´N, (2) C10, and (3) C2-meta-benzoate positions of taxane derivatives on their activity against sensitive versus counterpart paclitaxel-resistant breast (MCF-7) and ovarian (SK-OV-3) cancer cells. We found that (1) non-aromatic groups at both C3´ and C3´N positions, when compared with phenyl groups at the same positions of a taxane derivative, significantly reduced the resistance of ABCB1 expressing MCF-7/PacR and SK-OV-3/PacR cancer cells. This is, at least in the case of the SB-T-1216 series, accompanied by an ineffective decrease of intracellular levels in MCF-7/PacR cells. The low binding affinity of SB-T-1216 in the ABCB1 binding cavity can elucidate these effects. (2) Cyclopropanecarbonyl group at the C10 position, when compared with the H atom, seems to increase the potency and capability of the derivative in overcoming paclitaxel resistance in both models. (3) Derivatives with fluorine and methyl substituents at the C2-meta-benzoate position were variously potent against sensitive and resistant cancer cells. All C2 derivatives were less capable of overcoming acquired resistance to paclitaxel in vitro than non-substituted analogs. Notably, fluorine derivatives SB-T-121205 and 121,206 were more potent against sensitive and resistant SK-OV-3 cells, and derivatives SB-T-121405 and 121,406 were more potent against sensitive and resistant MCF-7 cells. (4) The various structure-activity relationships of SB-T derivatives observed in two cell line models known to express ABCB1 favor their complex interaction not based solely on ABCB1.

Aucubin directly targets β-catenin to co-suppress Wnt and HIF-1 pathways in ovarian cancer: Computational and experimental validation

OV is a highly lethal malignancy plagued by chemoresistance, necessitating the development of novel therapeutic agents. The Wnt/β-catenin pathway, driven by its central component β-catenin (CTNNB1), is a key oncogenic axis in OV, making it an attractive therapeutic target. We employed an integrated strategy combining in vitro assays (MTT, Western blot), in silico analyses (network pharmacology, WGCNA, molecular docking, molecular dynamics simulations), and biophysical validation assays (CETSA, DARTS) to systematically investigate the anti-cancer mechanism of the natural compound AU. AU potently inhibited the viability of A2780 OV cells by inducing apoptosis. Our comprehensive bioinformatic analysis identified CTNNB1 as a high-confidence direct target of AU. Clinical data confirmed that CTNNB1 is overexpressed in OV and correlates with poor patient prognosis. Molecular docking and dynamics simulations predicted a stable AU-CTNNB1 interaction, which was then experimentally validated by both CETSA and DARTS, confirming direct target engagement in a cellular context. Mechanistically, AU treatment resulted in the dose-dependent suppression of key proteins in the Wnt/β-catenin and HIF-1 signaling pathways. AU exerts its anti-OV activity by directly binding to CTNNB1. This interaction inhibits the oncogenic Wnt/β-catenin pathway, leading to the concurrent suppression of the HIF-1 pathway and the induction of apoptosis. Our study provides a complete mechanistic rationale for the development of AU as a novel targeted therapy for ovarian cancer.

Delivery of sPD1 gene by anti-CD133 antibody conjugated microbubbles combined with ultrasound for the treatment of cervical cancer in mice

To explore new therapeutic options for cervical cancer, the inhibitory effect on cervical cancer of targeted CD133-loaded sPD1 gene microbubbles (MBs) combined with low-frequency ultrasound was studied and its mechanism was explored. We prepared microbubbles conjugated with anti-CD133 antibody to deliver the sPD1 gene and determined concentration, particle size, and potentials of MBs. In addition, we verified that CD133 targeted-MBs could specifically bind to U14 cervical cancer cells in vitro. A mouse model of subcutaneous xenograft cervical cancer was established and mice were divided into a control group, an non-targeted microbubble group, a CD133-MBs group, an sPD1-MBs group and a CD133/sPD1-MBs group. Compared with the control group, tumor growth was inhibited in each group, with the CD133/sPD1 group showing the strongest inhibitory effect after treatment. The tumor volume and weight inhibition rates in the CD133/sPD1-MBs group were 78.01% and 72.25% respectively, which were statistically different from the other groups (P < 0.05), and HE staining and TUNEL immunofluorescence showed necrosis and apoptosis in tumor tissue. Flow cytometry, lactate dehydrogenase, and indirect immunofluorescence experiments showed that T lymphocytes were activated and a large number of CD8-positive T cells infiltrated the tumor tissue after treatment, with the CD133/sPD1-MBs group showing the most prominent effects (P < 0.05). The combination of ultrasound with anti- CD133 antibody-conjugated microbubbles loaded with the sPD1 gene can inhibit the growth of cervical cancer, suggesting that the immunosuppressive microenvironment of the tumor is improved after treatment.

Ganoderic acid T, a Ganoderma triterpenoid, modulates the tumor microenvironment and enhances the chemotherapy and immunotherapy efficacy through downregulating galectin-1 levels

Ganoderic acid T (GAT), a triterpenoid molecule of Ganoderma lucidum, exhibits anti-cancer activity; however, the underlying mechanisms remain unclear. Therefore, in this study, we aimed to investigate the anti-cancer molecular mechanisms of GAT and explore its therapeutic applications for cancer treatment. GAT exhibited potent anti-cancer activity in an ES-2 orthotopic ovarian cancer model in a humanized mouse model, leading to significant alterations in the tumor microenvironment (TME). Specifically, GAT reduced the proportion of α-SMA