Human Liver Cancer Research Articles

Characterization of an Mtbp Hypomorphic Allele in a Diethylnitrosamine-Induced Liver Carcinogenesis Model.

MTBP is implicated in cell cycle progression, DNA replication, and cancer metastasis. However, the function of MTBP remains enigmatic and is dependent on cellular contexts and its cellular localization. To understand the in vivo physiological role of MTBP, it is important to generate Mtbp knockout mice. However, complete deletion of the Mtbp gene in mice results in early embryonic lethality, while its heterozygous deletion shows modest biological phenotypes, including enhanced cancer metastasis. To overcome this and better characterize the in vivo physiological function of MTBP, we, for the first time, generated mice that carry an Mtbp hypomorphic allele (MtbpH) in which Mtbp protein is expressed at approximately 30% of that in the wild-type allele. We treated wild-type, Mtbp+/-, and MtbpH/- mice with a liver carcinogen, diethylnitrosamine (DEN), and found that the MtbpH/- mice showed worse overall survival when compared to the wild-type mice. Consistent with previous reports using human liver cancer cells, mouse embryonic fibroblasts (MEFs) from the MtbpH/- mice showed an increase in the nuclear localization of p-Erk1/2 and migratory potential. Thus, MtbpH/- mice and cells from MtbpH/- mice are valuable to understand the in vivo physiological role of Mtbp and validate the diverse functions of MTBP that have been observed in human cells.

Constitutive photomorphogenic protein 1 ubiquitinates interleukin-1 receptor accessory protein in human liver cancer.

Constitutive photomorphogenic protein 1 (COP1) plays a pivotal role in the development and progression of several human cancers and is reported to be upregulated in liver cancer. However, the role of COP1 in human liver cancer is unclear. We analyzed the COP1 expression in normal liver and liver cancer tissue samples using western blot and immunohistochemical analysis. We overexpressed and silenced COP1 in HepG2 and Huh7 cells and analyzed the effect on liver cancer cell proliferation. Additionally, COP1 was used as a bait to screen COP1-interacting proteins in a human cDNA library in a yeast two-hybrid screen and the results were confirmed with co-immunoprecipitation (co-IP) assays. Moreover, immunofluorescence staining was performed to assess co-localization. The protein levels of COP1 and mIL1RAcP were determined in clinical samples. COP1 was upregulated in liver cancer samples compared to that in normal tissue samples. COP1 overexpression promoted proliferation of liver cancer cells, while COP1 knockdown exerted the opposite effect. Yeast two-hybrid screen identified interleukin-1 receptor accessory protein (IL1RAP) as a potential COP1-interacting protein. Co-IP assays further confirmed that COP1 interacts with both preIL1RAP and membrane-bound form of IL1RAP (mIL1RAP). Furthermore, COP1 upregulated mIL1RAP protein levels and promoted nuclear translocation and activation of the nuclear factor kappa B (NF-κB) (p50/p65) dimer. Additionally, we demonstrated that COP1 regulated mIL1RAP expression through K63-linked polyubiquitination, suggesting that COP1 plays a role in stabilizing mIL1RAP. Finally, the protein levels of COP1 and mIL1RAcP were found to be positively correlated in clinical samples. COP1 regulates IL1RAP, which in turn results in activation of the NF-κB signaling. Our findings suggest that the COP1/IL1RAP/NF-κB axis promotes proliferation of liver cancer cells and is a potential target for the treatment of liver cancer.

PYCR2 promotes growth and aerobic glycolysis in human liver cancer by regulating the AKT signaling pathway

Hepatocellular carcinoma (HCC) is the world's third most fatal cancer. Because metabolic rewiring is a hallmark of HCC, studies into the causes of aberrant glycolysis could provide insight into novel HCC therapeutic strategies. Pyrroline-5-carboxylate reductase 2 (PYCR2), a key enzyme of proline synthesis, has previously been found to play vital roles in various malignancies regarding amino acid metabolism and oxidative stress response. Our study investigated the mechanistic function of PYCR2 in HCC. We used Gene Expression Profiling Interactive Analysis to perform bioinformatics analysis of PYCR2 expression and survival in human HCC patients based on the Cancer Genome Atlas database. The function of PYCR2 in cell viability and glycolysis was assessed using CCK-8 and ECAR assays. Transducing shRNA or overexpression vectors into the HCC cell line altered the expression status of PYCR2. PYCR2 expression was validated using quantitative real-time PCR and Western blot. In mouse xenograft models, the role of PYCR2 in HCC tumor formation was confirmed. PYCR2 was overexpressed in human HCC tumor tissue and was associated with a poor prognosis. The functional assay revealed that silencing PYCR2 inhibited cell viability, glycolysis, and AKT activation. Furthermore, the xenograft experiment demonstrated that silencing PYCR2 significantly inhibited tumor growth and Ki67 expression. On the other hand, PYCR2 overexpression significantly promoted cell viability and glycolysis, which could be inhibited by either a glycolysis inhibitor or an AKT inhibitor, indicating that PYCR2 may function via glycolysis and the AKT pathway. Moreover, despite the overexpression of PYCR2 in vivo, treatment with a glycolysis inhibitor may considerably suppress tumor growth. Our findings suggest that PYCR2 may play an oncogenic role in HCC growth by promoting glycolysis and activating AKT, emphasizing PYCR2's clinical relevance in HCC management as a novel potential therapeutic target.

Chemo-/Regio-Selective Synthesis of Novel Functionalized Spiro[pyrrolidine-2,3'-oxindoles] under Microwave Irradiation and Their Anticancer Activity.

A novel series of nitrostyrene-based spirooxindoles were synthesized via the reaction of substituted isatins 1a-b, a number of α-amino acids 2a-e and (E)-2-aryl-1-nitroethenes 3a-e in a chemo/regio-selective manner using [3+2] cycloaddition (Huisgen) reaction under microwave irradiation conditions. The structure elucidation of all the synthesized spirooxindoles were done using 1H and 13C NMR and HRMS spectral analysis. The single crystal X-ray crystallographic study of compound 4l was used to assign the stereochemical arrangements of the groups around the pyrrolidine ring in spiro[pyrrolidine-2,3'-oxindoles] skeleton. The in vitro anticancer activity of spiro[pyrrolidine-2,3'-oxindoles] analogs 4a-w against human lung (A549) and liver (HepG2) cancer cell lines along with immortalized normal lung (BEAS-2B) and liver (LO2) cell lines shows promising results. Out of the 23 synthesized spiro[pyrrolidine-2,3'-oxindoles], while five compounds (4c, 4f, 4m, 4q, 4t) (IC50 = 34.99-47.92 µM; SI = 0.96-2.43) displayed significant in vitro anticancer activity against human lung (A549) cancer cell lines, six compounds (4c, 4f, 4k, 4m, 4q, 4t) (IC50 = 41.56-86.53 µM; SI = 0.49-0.99) displayed promising in vitro anticancer activity against human liver (HepG2) cancer cell lines. In the case of lung (A549) cancer cell lines, these compounds were recognized to be more efficient and selective than standard reference artemisinin (IC50 = 100 µM) and chloroquine (IC50 = 100 µM; SI: 0.03). However, none of them were found to be active as compared to artesunic acid [IC50 = 9.85 µM; SI = 0.76 against lung (A549) cancer cell line and IC50 = 4.09 µM; SI = 2.01 against liver (HepG2) cancer cell line].

Green-fabricated MgO nanoparticles: A potent antimicrobial and anticancer agent

MgO nanoparticles are gaining popularity because of their potential applications in several different industries, such as bioengineering, medicine, and environmental protection. The fabrication of MgO nanoparticles with enhanced biological properties remains difficult, despite the growing interest in this area. In this paper, we describe a method that minimizes environmental impact when manufacturing MgO nanoparticles using garlic (Allium sativum L.) extract. For ages, one of the most vital functions in human nutrition and medicine has been played by garlic. Garlic also contains phytochemicals that are effective against cancer and microbes. Our goal was to increase the medicinal relevance of MgO nanoparticles while decreasing our reliance on harmful chemicals. Various characterization techniques were employed to confirm the synthesis of MgO nanoparticles, including SEM, TEM, EDX, and XRD. The study findings indicate that the synthesis of MgO nanoparticles yielded a polycrystalline cubic configuration with a crystal size of 55–60 nm. The results of the EDX analysis suggest that only Mg and O elements are present, without any detectable impurities. The efficacy of MgO nanoparticles against gram-positive (Staphylococcus epidermidis), gram-negative bacteria (Escherichia coli) and a fungal pathogen (Candida albicans) was investigated. The results indicated that MgO nanoparticles exhibited high effectiveness against all three microorganisms. The study revealed that MgO nanoparticles exhibit potent anticancer properties against human liver (HepG2) and lung (A549) cancer cells. Reactive oxygen species production in cancer cells by MgO nanoparticles suggests oxidative stress pathway-mediated anticancer action. The present research highlights the significance of utilizing medicinal plants in the synthesis of nanoparticles for enhanced anticancer and microcidal properties.

Facile phytosynthesis of gold nanoparticles using Nepeta bodeana Bunge: Evaluation of its therapeutics and potential catalytic activities

In the present study, the ethanol extract of leaves of Nepeta bodeana Bunge was utilized in environmentally friendly synthesis of gold nanoparticles (AuNPs) via an immediate, one-step, and cost-effective process. The biosynthesized AuNPs were characterized using a variety of techniques, including UV–visible spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), high-resolution transmission electron microscopy (HRTEM), and laser Doppler electrophoresis (zeta potential), among others. Pink in hue, the manufactured AuNPs were practically spherical and had<20.5 nm in diameter. The peak of the localized surface plasmon resonance absorption band at 547 nm helped to explain the biogenic synthesis of AuNPs, which used the ethanolic extract of the Nepeta bodeana leaves. In addition to this, the biosynthesized AuNPs possessed the remarkable stability as a result of their zeta potential value of − 22.3 mV. The green synthesized AuNPs demonstrated the excellent catalytic activity because they were able to catalytically degrade methylene blue (MB) at a variety of concentrations. The results indicated that 94.2% of MB were degraded after 10 min. They also showed a remarkable antibacterial activity against both Gram-nrgative (Pseudomonas aeruginosa and Escherichia coli) and Gram-positive (Staphylococcus aureus and Bacillus cereus) bacteria. In addition, a proper antifungal activity with the minimum inhibitory concentration (MIC) values of 50 and 25 μg/mL against Candida albicans and Aspergillus niger, respectively, were determined. The phytosynthesized AuNPs indicated a considerable antioxidant activity of 83% and 74% in the applied DPPH and ABTS radical scavenging activity tests, respectively. The anti-inflammatory properties of the synthesized AuNPs were established to be dose-dependent and varied from 11% to 76%. The green synthesized AuNPs also exhibited a concentration-dependent cytotoxicity on human liver (HepG2) and breast (MCF-7) cancer cells, which were around 77.4 and 71.2% at 300 µg/mL, respectively. They showed as well a notable anti-diabetic activity with the 52% inhibition of α-amylase and the 62% glucose uptake activity by yeast cells at 300 µg/mL. All these findings suggested that the produced AuNPs could be explored for developing many therapeutic as well as catalytic useful products.

In vitro anticancer potential of laminarin and fucoidan from Brown seaweeds

Marine seaweeds are rich source of polysaccharides present in their cell wall and are cultivated and consumed in China, Japan, Korea, and South Asian countries. Brown seaweeds (Phaeophyta) are rich source of polysaccharides such as Laminarin and Fucoidan. In present study, both the laminarin and fucoidan were isolated was yielded higher in PP (Padina pavonica) (4.36%) and STM (Stoechospermum marginatum) (2.32%), respectively. The carbohydrate content in laminarin and fucoidan was 86.91% and 87.36%, whereas the sulphate content in fucoidan was 20.68%. Glucose and mannose were the major monosaccharide units in laminarin (PP), however, fucose, galactose, and xylose in fucoidan (STM). FT-IR down peaks represent the carbohydrate of laminarin and fucoidan except, for 1219 cm−1, and 843 cm−1, illustrating the sulphate groups of fucoidan. The molecular weight of laminarin was 3–5 kDa, and the same for fucoidan was 2–6 kDa, respectively. Both the Fucoidan and Laminarin showed null cytotoxicity on Vero cells. Contrastingly, the fucoidan possess cytotoxic activity on human liver cancer cells (HepG2) (IC50—24.4 ± 1.5 µg/mL). Simultaneously, laminarin also shown cytotoxicity on human colon cancer cells (HT-29) (IC50—57 ± 1.2 µg/mL). The AO/EB (Acriding Orange/Ethidium Bromide) assay significantly resulted in apoptosis and necrosis upon laminarin and fucoidan treatments, respectively. The DNA fragmentation results support necrotic cancer cell death. Therefore, laminarin and fucoidan from PP and STM were potential bioactive compounds for anticancer therapy.

Dual and triple encapsulated iron gluconate speed up anemia recovery in an animal model

Food fortification can be a solution to anemia in developing countries. A previous study determined that the combination of spray drying, hydroxypropyl-methylcellulose as wall material, and maltodextrin as bulk material, encapsulated iron gluconate achieved the highest bioavailability. However, the addition of vitamin B12 to the hydroxypropyl-methylcellulose/maltodextrin capsules increased the iron cell uptake over the previously reported results. The cell viability, the number of live, healthy cells in a sample, of HepG2, human liver cancer cells, increases by about 17% for dual-encapsulated iron gluconate and vitamin b12. The cell uptake in Caco2, human colorectal adenocarcinoma cells, is higher by 25% when using encapsulated iron and vitamin b12 compared to encapsulated iron. The strength of dual-encapsulated iron and vitamin b12 is also confirmed in in-vivo studies. Once fully anemic, young female rats eating food with encapsulated iron gluconate and vitamin b12, show the fastest recovery with respect to rats eating food with encapsulated iron and pure iron. The first needed only five days for their hemoglobulin values to return to normal. The second and the third needed 15 and 21 days, respectively.

Investigation of In Vitro antiproliferative activity properties of Spartium junceum L. (Spanish broom) against MDA-MB-231 and HepG2 cancer cell lines

Cancer is among the top global public health burdens leading to millions of deaths each year. The study aims to investigate the antiproliferative effect of Spartium junceum L. flowers on different cancer cell lines. The ethanolic extract of the flowers was prepared in the present study. Phytochemical analysis of the plant extract revealed the presence of several phenolic compounds such as cinnamic acid and its derivatives (chlorogenic, p-coumaric, ferulic acids), protocatechuic acid, epicatechin and luteolin. This extract was tested against human breast (MDA-MB-231) and liver (HepG2) cancer cell lines to find out its antiproliferative activity. It was determined that the extract was effective against both cell lines with IC50 values of 2.37 ± 0.47 and 0.98 ± 0.01 µL/mL for MDA-MB-231 and HepG2, respectively. Particularly, the extract was found to be more effective in the liver cancer cell line than the breast cancer cell line. All these obtained findings led us to believe that this medicinal plant could be a promising antiproliferative agent candidate for the treatment of human liver and breast cancers.

Oncogenic β-catenin-driven liver cancer is susceptible to methotrexate-mediated disruption of nucleotide synthesis.

Liver cancer is largely resistant to chemotherapy. This study aimed to identify the effective chemotherapeutics for β-catenin-activated liver cancer which is caused by gain-of-function mutation of catenin beta 1 ( CTNNB1 ), the most frequently altered proto-oncogene in hepatic neoplasms. Constitutive β-catenin-activated mouse embryonic fibroblasts (MEFs) were established by deleting exon 3 ( β-catenin Δ(ex3)/+ ), the most common mutation site in CTNNB1 gene. A screening of 12 widely used chemotherapy drugs was conducted for the ones that selectively inhibited β-catenin Δ(ex3)/+ but not for wild-type MEFs. Untargeted metabolomics was carried out to examine the alterations of metabolites in nucleotide synthesis. The efficacy and selectivity of methotrexate (MTX) on β-catenin-activated human liver cancer cells were determined in vitro . Immuno-deficient nude mice subcutaneously inoculated with β-catenin wild-type or mutant liver cancer cells and hepatitis B virus ( HBV ); β-catenin lox(ex3)/+ mice were used, respectively, to evaluate the efficacy of MTX in the treatment of β-catenin mutant liver cancer. MTX was identified and validated as a preferential agent against the proliferation and tumor formation of β-catenin-activated cells. Boosted nucleotide synthesis was the major metabolic aberration in β-catenin-active cells, and this alteration was also the target of MTX. Moreover, MTX abrogated hepatocarcinogenesis of HBV ; β-catenin lox(ex3)/+ mice, which stimulated concurrent Ctnnb1- activated mutation and HBV infection in liver cancer. MTX is a promising chemotherapeutic agent for β-catenin hyperactive liver cancer. Since repurposing MTX has the advantages of lower risk, shorter timelines, and less investment in drug discovery and development, a clinical trial is warranted to test its efficacy in the treatment of β-catenin mutant liver cancer.

Anti-cancer effects of the extracts of broad and spirale cultivars of Codiaeum variegatum (L.) Blume on MCF-7, HepG2, and HeLa cell lines

Introduction: Codiaeum variegatum (L.) Blume is a well-known ornamental foliage plant used as a vegetable in northern Thailand, and it is the source of numerous bioactive substances. This work explored the effects of leaf extracts of broad (BCE) and spirale (SCE) cultivars of C. variegatum on three cancer cells, including human breast, human liver, and human cervical cancer cells. Methods: Ethanolic plant extracts were prepared, and then, 2,2-diphenyl-1-picryhydrazyl (DPPH), ferrous iron chelating, and lipid peroxidation assays were used to examine the flavonoid and phenolic compounds. The proliferative inhibition, growth, and migration of MCF-7, HepG2, and HeLa cancer cells, as a result of exposure to the extracts, were investigated. The extracts were investigated for their anti-cancer activities using sulforhodamine B (SRB), clonogenic, and wound-healing methods. Results: The data demonstrated that BCE and SCE contained high phenolic compounds. However, both extracts showed inactive anti-oxidant activities. Both extracts had high cytotoxicity on three types of cancer cells in a dose- and time-dependent manner after 24-72 hours of incubation with IC50 values in a range of 208-830 µg/mL. Moreover, the prepared extracts of C. variegatum significantly inhibited colony-forming ability and cell migration on all types of cancer cells. Compared with BCE, SCE showed more potent anti-cancer activities. Conclusion: These findings revealed that SCE had higher anti-cancer activities on MCF-7, HepG2, and HeLa cancer cells than BCE. Consequently, the SCE might be used as an effective chemotherapeutic compound for the prevention and treatment of cancer.

Chemical composition and cytotoxicity of Salicornia europaea L

Salicornia europaea L. is an economic plant which can grow in saline-alkali land, beach and other heavy saline soil. In this study, the inhibitory activities of S. europaea extracts against tumor cell growth including human non-small cell lung cancer cell A549, human liver cancer cell HepG2, and brain glioma cell U251 were screened. The results showed that the ethanol extract of S. europaea had an inhibitory effect on the proliferation of A549 cells, and the inhibition of tumor cells showed time-dose relationship. Moreover, the ethanol extract of S. europaea was extracted and fractionated with different organic solvents and the activity results indicated that the petroleum ether fraction with IC50 of 96.5 μg mL−1 possessed the best activity against A549 cells. Then seven compounds, including 30-norhederagenin (1), 24-methylenecycoartanol (2), pheophorbide A (3), stigmasterol (4), β-sitosterol (5), α-spinasterol (6), phytol (7) were isolated and identified from the petroleum ether fraction of S. europaea. Besides, the oil obtained from the petroleum ether fraction was analyzed by GC-MS, and fourteen compounds were identified. The compounds identified from the petroleum ether fraction mainly consisted of triterpenes, sterols, fatty acids, and their lipids.

Anti-PD-L1 antibodies promote cellular proliferation by activating the PD-L1-AXL signal relay in liver cancer cells.

Immune checkpoint inhibitors (ICIs) are emerging treatments for advanced hepatocellular carcinoma (HCC); however, evidence has shown they may induce hyperprogressive disease via unexplained mechanisms. In this study, we investigated the possible stimulative effect of ICIs on programmed cell death-ligand 1 (PD-L1)-harboring liver cancer cells under immunocompetent cell-free conditions. The sarcomatous HAK-5 cell line displayed the highest expression of PD-L1 among 11 human liver cancer cell lines used in this study. HLF showed moderate expression, while HepG2, Hep3B, and HuH-7 did not show any. Moreover, sarcomatous HCC tissues expressed high levels of PD-L1. We observed approximately 20% increase in cell proliferation in HAK-5 cells treated with anti-PD-L1 antibodies, such as durvalumab and atezolizumab, for 48h compared with that of those treated with the control IgG and the anti-PD-1 antibody pembrolizumab. No response to durvalumab or atezolizumab was shown in PD-L1-nonexpressing cells. Loss-of-function and gain-of-function experiments for PD-L1 in HAK-5 and HepG2 cells resulted in a significant decrease and increase in cell proliferation, respectively. Phosphorylated receptor tyrosine kinase array and immunoprecipitation revealed direct interactions between PD-L1 and AXL in tumor cells. This was stabilized by extrinsic anti-PD-L1 antibodies in a glycosylated PD-L1-dependent manner. Activation of AXL, triggering signal relay to the Akt and Erk pathways, boosted tumor cell proliferation both in vitro and in xenografted tumors in NOD/SCID mice. Collectively, this suggests that anti-PD-L1 antibodies stimulate cell proliferation via stabilization of the PD-L1-AXL complex in specific types of liver cancer, including in HCC with mesenchymal components. Therapeutic anti-PD-L1 antibodies promote cell proliferation by stabilizing the PD-L1-AXL complex in PD-L1-abundant neoplasms, including in HCC with mesenchymal components. Such a mechanism may contribute to the development of hyperprogressive disease.

SONAR enables cell type deconvolution with spatially weighted Poisson-Gamma model for spatial transcriptomics

Recent advancements in spatial transcriptomic technologies have enabled the measurement of whole transcriptome profiles with preserved spatial context. However, limited by spatial resolution, the measured expressions at each spot are often from a mixture of multiple cells. Computational deconvolution methods designed for spatial transcriptomic data rarely make use of the valuable spatial information as well as the neighboring similarity information. Here, we propose SONAR, a Spatially weighted pOissoN-gAmma Regression model for cell-type deconvolution with spatial transcriptomic data. SONAR directly models the raw counts of spatial transcriptomic data and applies a geographically weighted regression framework that incorporates neighboring information to enhance local estimation of regional cell type composition. In addition, SONAR applies an additional elastic weighting step to adaptively filter dissimilar neighbors, which effectively prevents the introduction of local estimation bias in transition regions with sharp boundaries. We demonstrate the performance of SONAR over other state-of-the-art methods on synthetic data with various spatial patterns. We find that SONAR can accurately map region-specific cell types in real spatial transcriptomic data including mouse brain, human heart and human pancreatic ductal adenocarcinoma. We further show that SONAR can reveal the detailed distributions and fine-grained co-localization of immune cells within the microenvironment at the tumor-normal tissue margin in human liver cancer.

Modifications of The Human Liver Cancer Cells through microRNA-145-Mediated Targeting of CDCA3.

Owing to the lethality of liver cancer, it is considered as one of the devastating types of cancers across the globe. Consistently, the study was designed to elucidate the role and to explore the therapeutic implications of miR-145 in human liver cancer. In the current experimental study, gene expression was determined by RT-PCR analysis. Transfection of cancer cells was carried out using Lipofectamine 2000. The cell proliferation of liver cancer cells was estimated by MTT assay. Clonogenic assay was performed for analysis of colony forming potential of cancer cells. Flow cytometry was done to analyze the cell cycle phase distribution of cancer cells. Transwell chamber assay was performed to assess the motility of cancer cells. Western blotting was done to estimate the expression levels of proteins. Dual luciferase assay was performed for interaction analysis of miR-145 with CDCA3. The miR-145 expression was found to be downregulated in liver cancer cells. The transfection mediated overexpression of miR-145 inhibited the cancer cell proliferation and when miR-145 inhibitor was transfected, cancer cells showed higher proliferation rates. Enrichment of miR-145 levels led to cell cycle arrest at G2/M phase by inhibiting cyclin B1. miR-145 also restricted the migration and invasion of cancer cells. CDCA3 was shown to be the intracellular target of miR-145 and it was found that the inhibitory effects of miR-145 were modulated through CDCA3, intracellularly. The current study clearly revealed that there is a need to investigate the regulatory role of different molecular entities like microRNAs in cancer development to better understand mechanics behind this pathogenesis and design more effective combating strategies against cancer.

Curcumin promotes apoptosis of liver cancer cells by down-regulating WNT/β-catenin

Curcumin (C21H20O6) is considered to exert anticancer potential. However, the relationship between curcumin and β-catenin has not been fully clarified and its regulatory role in human liver cancer has not yet been confirmed. In this study, human liver cancer cells were exposed to curcumin and the proliferation of HepG2 cells was detected by MTT method along with analysis of cell cycle and apoptosis, as well as Wnt/β-catenin signaling proteins expression. Curcumin shows a time-dependent and dose-dependent effect on HepG2 cell proliferation. The IC50 at 24 h, 48 h and 72 h were 12.8±0.67) μmol/L, 8.8±0.43 μmol/L and 4.6±0.42 μmol/L, respectively. Curcumin could dose-dependently (5, 10 and 20 μmol/L) arrest HepG2 cells in the G2/M phase (p &lt; 0.05) and increase apoptosis (p &lt; 0.05). In addition, curcumin down-regulated total, cytoplasmic and nuclear β-catenin proteins in HepG2 cells, indicating inhibited Wnt/β-catenin signaling. Curcumin reduced the expression of Dvl-2, Dvl-3, GSK-3β (p-ser9), C-myc, and Survivin, and increased the expression of GSK-3 (p-tyr216) and Axin-2 without affecting total GSK-3β levels. Curcumin is able to inhibit liver cancer cell activities possibly through inhibition of Wnt/β-catenin signaling.

Apoptosis induction in human hepatoma cell line HepG2 cells by trans- Anethole via activation of mitochondria-mediated apoptotic pathways

trans-Anethole a valuable compound derived from star anise widely used by ethnic tribals to manage numerous human diseases. In this study antiproliferative activities of trans-Anethole towards human liver cancer (HepG2), cervical cancer (HeLa) and breast cancer (MCF-7) cells were explored. trans-Anethole showed free radical scavenging potential as assessed by DNA nicking assay. trans-Anethole exhibited strong antiproliferative potential towards HepG2 cells compared to other cell lines. trans-Anethole strongly induced apoptosis in HepG2 cells by significantly upregulating the protein expressions of p53, Caspase-3 and Caspase-9 were assessed by western blotting analysis which highlighted apoptosis-inducing capacity of trans-Anethole against HepG2 cells. Rt-qPCR analysis revealed that trans- Anethole upregulated p53, caspase − 3 and − 9 in comparison to untreated HepG2 cancer cells. Moreover, trans-Anethole provoked the generation of ROS and disruption of MMP. Our research suggests that trans-Anethole may have a significant anticancer therapeutic potential for treating liver cancer.

Uncovering the Gut-Liver Axis Biomarkers for Predicting Metabolic Burden in Mice.

Western diet (WD) intake, aging, and inactivation of farnesoid X receptor (FXR) are risk factors for metabolic and chronic inflammation-related health issues ranging from metabolic dysfunction-associated steatotic liver disease (MASLD) to dementia. The progression of MASLD can be escalated when those risks are combined. Inactivation of FXR, the receptor for bile acid (BA), is cancer prone in both humans and mice. The current study used multi-omics including hepatic transcripts, liver, serum, and urine metabolites, hepatic BAs, as well as gut microbiota from mouse models to classify those risks using machine learning. A linear support vector machine with K-fold cross-validation was used for classification and feature selection. We have identified that increased urine sucrose alone achieved 91% accuracy in predicting WD intake. Hepatic lithocholic acid and serum pyruvate had 100% and 95% accuracy, respectively, to classify age. Urine metabolites (decreased creatinine and taurine as well as increased succinate) or increased gut bacteria (Dorea, Dehalobacterium, and Oscillospira) could predict FXR deactivation with greater than 90% accuracy. Human disease relevance is partly revealed using the metabolite-disease interaction network. Transcriptomics data were also compared with the human liver disease datasets. WD-reduced hepatic Cyp39a1 (cytochrome P450 family 39 subfamily a member 1) and increased Gramd1b (GRAM domain containing 1B) were also changed in human liver cancer and metabolic liver disease, respectively. Together, our data contribute to the identification of noninvasive biomarkers within the gut-liver axis to predict metabolic status.

Nickel chloride complexes with substituted 4'-phenyl-2',2':6',2″-terpyridine ligands: synthesis, characterization, anti-proliferation activity and biomolecule interactions.

A series of Ni(II) sandwich-like coordinated compounds were synthesized by the reaction of nickel dichloride and ten 4'-(4-substituent phenyl)-2',2':6',2″-terpyridine ligands, and their structures were confirmed by elemental analysis, FT-IR, ESI-MS, solid state ultraviolet spectroscopy and X-ray single crystal diffraction analysis. Three human cancer cell lines and a normal human cell line were used for anti-proliferation potential study: human lung cancer cell line (A549), human esophageal cancer cell line (Eca-109), human liver cancer cells (Bel-7402) and normal human liver cells (HL-7702). The results show that these nickel complexes possess good inhibitory effects on the cancer cells, outperforming the commonly used clinical chemotherapy drug cisplatin. Especially, complexes 3 (-methoxyl) and 7 (-fluoro) have strong inhibitory ability against Eca-109 cell line with IC50 values of 0.223μM and 0.335μM, complexes 4 and 6 showed certain cell selectivity, and complex 6 can inhibit cancer cells and slightly poison normal cells when the concentration was controlled. The ability of these complexes binding to CT-DNA was studied by UV titration and CD spectroscopy, and CD spectroscopy was also used to study the secondary structural change of BSA under the action of the complexes. The binding of these complexes with DNA, DNA-Topo I and bovine serum protein has been simulated by molecular docking software, and the docking results and optimal binding conformation data showed that they interacted with DNA in the mode of embedded binding, which is consistent with the experimental results. These complexes are more inclined to move to the cleavage site when docking with DNA-Topo I, so as to play a role of enzyme cleavage, while BSA promotes the action of the complexes by binding to effective binding sites.

Microtubule-Associated Serine/Threonine (MAST) Kinases in Development and Disease.

Microtubule-Associated Serine/Threonine (MAST) kinases represent an evolutionary conserved branch of the AGC protein kinase superfamily in the kinome. Since the discovery of the founding member, MAST2, in 1993, three additional family members have been identified in mammals and found to be broadly expressed across various tissues, including the brain, heart, lung, liver, intestine and kidney. The study of MAST kinases is highly relevant for unraveling the molecular basis of a wide range of different human diseases, including breast and liver cancer, myeloma, inflammatory bowel disease, cystic fibrosis and various neuronal disorders. Despite several reports on potential substrates and binding partners of MAST kinases, the molecular mechanisms that would explain their involvement in human diseases remain rather obscure. This review will summarize data on the structure, biochemistry and cell and molecular biology of MAST kinases in the context of biomedical research as well as organismal model systems in order to provide a current profile of this field.