Growth Of Liver Cancer Cells Research Articles

Inhibition of NPC1 suppresses cell proliferation and β-catenin signaling activation of liver cancer.

Niemann-Pick Type C1 (NPC1) plays a significant role in the development of liver diseases and liver cancer. Our objective was to investigate the involvement of NPC1 in regulating liver cancer development. We observed that high levels of NPC1 expression in tumor tissues from patients with liver cancer were associated with a poor prognosis. Through in vitro experiments, we found that inhibiting NPC1 expression reduced the proliferation, invasion, and migration of liver cancer cells, while also inducing apoptosis in these cells. Additionally, the inhibition of NPC1 led to decreased activation of Wnt/β-catenin signaling. In vivo studies further supported our findings by demonstrating that the suppression of liver cancer cell growth was effectively achieved through the inhibition of NPC1. Overall, our results strongly indicate that the inhibition of NPC1 suppresses liver cancer cell proliferation. Targeting NPC1 is a promising potential therapeutic strategy for liver cancer.

The Effects of Crude Alkaloid Extracts of Convolutes Arvensis on Tumour Cell Lines and Their Potential For Toxicity

General Background: Cancer remains a leading cause of mortality worldwide, and the search for effective treatments continues to intensify. Specific Background: Convolvulus arvensis has been traditionally used for medicinal purposes, but its potential as an anticancer agent is underexplored. Knowledge Gap: The cytotoxic and apoptotic mechanisms of crude alkaloids extracted from C. arvensis against specific cancer cell lines have not been fully characterized. Aims: This study investigates the antioxidant activity and cytotoxic effects of C. arvensis crude alkaloids on mouse liver cancer (HC) and human breast cancer (AMJ13) cell lines, focusing on apoptosis-related gene expression. Results: The antioxidant activity of C. arvensis was comparable to that of ascorbic acid, with inhibition rates of 92.01% at 500 µg/ml. Crude alkaloids demonstrated dose-dependent cytotoxicity, with a maximum inhibition rate of 82.65% in AMJ13 cells and 79.49% in HC cells at 500 µg/ml. Gene expression analysis revealed upregulation of caspase-9, indicating apoptosis via the intrinsic mitochondrial pathway. Novelty: This study is among the first to provide molecular evidence of C. arvensis-induced apoptosis through the intrinsic pathway, offering a novel insight into its anticancer potential. Implications: These findings suggest that C. arvensis alkaloids could be developed as a therapeutic option for cancer treatment, with future studies needed to isolate specific compounds and assess their in vivo efficacy. Highlights: C. arvensis alkaloids show antioxidant activity similar to ascorbic acid. Alkaloids inhibit liver and breast cancer cell growth dose-dependently. Apoptosis triggered via caspase-9 through the mitochondrial pathway. Keywords: Convolvulus arvensis, alkaloids, cancer, apoptosis, antioxidant

Proanthocyanidin Regulates NETosis and Inhibits the Growth and Proliferation of Liver Cancer Cells - In Vivo, In Vitro and In Silico Investigation.

Liver cancer ranks third in global cancer-related mortality, with about 700,000 deaths recorded yearly, making it one of the most common cancers worldwide. Even though prognoses differ according to the severity of the diseases, many patients now exhibit an increased life cycle since the implementation of chemotherapy. In the current study, we investigated the effect of proanthocyanidin ‒a polyphenol molecule found in many plants‒ on the proliferation and invasion of liver cancer cells. In particular, we determined the effect of proanthocyanidin on the serum levels of four strategic liver cancer target, TNFα, IL-6, cfDNA, and IL-1β. Further molecular insight on the inhibitory mechanism of proanthocyanidin against TNFα, IL-6, and IL-1β was obtained via molecular docking, molecular dynamics simulations and binding free energy calculations. Results showed that proanthocyanidin inhibited the growth of HepG2 and HEP3B cells, and effectively reduced clonogenic survival and invasion potential when compared to control cells. Proanthocyanidin was also found to suppress the expression of Bcl-2 (26 kDa) protein in HepG2 cells, while increasing the expression of Bax (21 kDa). Molecular dynamics (MD) and thermodynamic binding free energy calculations showed that proanthocyanidin maintained stable binding within the active site of target proteins across the entire 100 ns MD simulation period, and its binding affinity outscored respective control molecules.In conclusion, the multifaceted analysis showcased in this study demonstrated promising anti-cancer effect of proanthocyanidin on HepG2 and HEP3B cancer cells, highlighting its potential as a viable liver cancer therapeutic alternative.

5149 In-Vitro Combo Treatment of Human Liver Cancer Cell (Hep-G2) Line With Vitamin-A And RU-486

Abstract Disclosure: P. Ramaraj: None. Our previous in-vitro studies involving vitamins (vitamin-A and vitamin-D3) and steroids (progesterone and RU-486) pointed to a combination of vitamin-A and RU-486 as an efficient combo to decrease human melanoma cell growth. We extended this combo treatment to human liver cancer cell (Hep-G2) line. The aim was to check whether vit-A and RU-486 combo would also decrease human liver cancer cell line growth. Liver cancer (Hep-G2) cells were plated in a 96 well plate overnight. Following day, vit-A and RU-486 were added either alone or in different combinations for 48 hrs. After 48 hrs, cell viability and percentage of cell growth were monitored by the MTT assay. Supernatants were collected for Elisarray and quantitation of cytokine(s) by Elisa. Individual vit-A and RU-486 treatments showed a dose-dependent decrease in cell growth. Though various combinations of vit-A (50, 75, 100 μM) and RU-486 (10, 50 100 μM) also showed a decrease in cell growth, an experimentally significant decrease in cell growth was observed at vit-A 75 μM and RU-486 50 μM combination. This combo resulted in 24% cell growth compared to straight vit-A (75 μM) at 44% cell growth and RU-486 (50 μM) at 35% cell growth. Thus, this combo inhibited liver cancer cell growth also. In the future effect of this combo on normal liver cell line and the cytokine(s) if any, playing a central role in cell growth would be identified by Elisarray and quantitated by Elisa. Presentation: 6/3/2024

5149 In-vitro Combo Treatment of Human Liver Cancer cell (Hep-G2) Line with Vitamin-A and RU-486

Abstract Disclosure: P. Ramaraj: None. Our previous in-vitro studies involving vitamins (vitamin-A and vitamin-D3) and steroids (progesterone and RU-486) pointed to a combination of vitamin-A and RU-486 as an efficient combo to decrease human melanoma cell growth. We extended this combo treatment to human liver cancer cell (Hep-G2) line. The aim was to check whether vit-A and RU-486 combo would also decrease human liver cancer cell line growth. Liver cancer (Hep-G2) cells were plated in a 96 well plate overnight. Following day, vit-A and RU-486 were added either alone or in different combinations for 48 hrs. After 48 hrs, cell viability and percentage of cell growth were monitored by the MTT assay. Supernatants were collected for Elisarray and quantitation of cytokine(s) by Elisa. Individual vit-A and RU-486 treatments showed a dose-dependent decrease in cell growth. Though various combinations of vit-A (50, 75, 100 μM) and RU-486 (10, 50 100 μM) also showed a decrease in cell growth, an experimentally significant decrease in cell growth was observed at vit-A 75 μM and RU-486 50 μM combination. This combo resulted in 24% cell growth compared to straight vit-A (75 μM) at 44% cell growth and RU-486 (50 μM) at 35% cell growth. Thus, this combo inhibited liver cancer cell growth also. In the future effect of this combo on normal liver cell line and the cytokine(s) if any, playing a central role in cell growth would be identified by Elisarray and quantitated by Elisa. Presentation: 6/3/2024

PYCR1 promotes liver cancer cell growth and metastasis by regulating IRS1 expression through lactylation modification.

Liver cancer (LC) is among the deadliest cancers worldwide, with existing treatments showing limited efficacy. This study aimed to elucidate the role and underlying mechanisms of pyrroline-5-carboxylate reductase 1 (PYCR1) as a potential therapeutic target in LC. Immunohistochemistry and Western blot were used to analyse the expression of PYCR1 in LC cells and tissues. EdU assays, colony-forming assays, scratch wound healing assays, Transwell assays, nude mouse xenograft models and nude mouse lung metastasis models were used to detect the growth and metastasis abilities of LC cells. Transcriptome sequencing was used to search for downstream target genes regulated by PYCR1, and metabolomics was used to identify the downstream metabolites regulated by PYCR1. ChIP assays were used to analyse the enrichment of H3K18 lactylation in the IRS1 promoter region. We found that the expression of PYCR1 was significantly increased in HCC and that this high expression was associated with poor prognosis in HCC patients. Knockout or inhibition of PYCR1 inhibited HCC cell proliferation, migration and invasion both in vivo and in vitro. In addition, we revealed that knocking out or inhibiting PYCR1 could inhibit glycolysis in HCC cells and reduce H3K18 lactylation of the IRS1 histone, thereby inhibiting IRS1 expression. Our findings identify PYCR1 as a pivotal regulator of LC progression that influences tumour cell metabolism and gene expression. By demonstrating the potential of targeting PYCR1 to inhibit LC cell proliferation and metastasis, this study identified PYCR1 as a promising therapeutic target for LC. Pyrroline-5-carboxylate reductase 1 (PYCR1) promotes the proliferation and metastasis of liver cancer (LC) cells. The expression of PYCR1 in LC is regulated by DNA methylation. Knocking down or inhibiting PYCR1 inhibits glycolysis as well as the PI3K/AKT/mTOR and MAPK/ERK pathways in LC cells. PYCR1 regulates the transcriptional activity of IRS1 by affecting H3K18 lactylation in its promoter region.

Synergistic chemo-photothermal anticancer effect of pH-responsive curcumin loaded mesoporous silica decorated reduced graphene

Synergistic chemo-photothermal therapy is employed to treat cancer. In this work, the curcumin loaded mesoporous silica on reduced graphene oxide (Cur-mSiO2@rGO) is synthesized. The mSiO2@rGO nanocarrier combines rGO to absorb NIR radiations with mSiO2 to load the drug. The mSiO2@rGO thus functions as bimodal photothermal agent and pH-responsive drug nanocarrier. Curcumin loaded onto the nanocarrier possesses antiproliferative, antioxidant, anti-inflammatory, pro-apoptotic, and antiangiogenic characteristics. It has anticancer potential against malignancies of stomach, liver, breast, lungs, and prostate. Breast cancer (BCa) is a major health concern worldwide. Curcumin inhibits BCa by mechanisms including apoptosis, cell cycle arrest, and modulation of signaling pathways. Moreover, curcumin blocks transcription factor (NF-κB), PI3K/AKT signaling, and STAT3 protein to inhibit liver cancer cell growth and survival. Cur-mSiO2@rGO targets cancer cells with enhanced loading capacity of 92 ± 1.02 % curcumin which is specifically released in the acidic tumor environment. NIR lamp irradiation at 808 nm ablates cancer cells to demonstrate the high photothermal conversion efficiency of rGO. In vitro experiments prove that synergistic chemo-photothermal therapy effectively kills cancer cells (HepG2 and MCF-7) compared to the single chemotherapeutic treatment. The toxicity of Cur-mSiO2@rGO is analyzed in vivo by the serum biochemical analysis, and biosafety by histopathological examination of vital body organs. A 1000 mg/kg specific dose of Cur-mSiO2@rGO shows no toxic effect on mice organs. Additionally, the in vivo studies confirm that Cur-mSiO2@rGO with NIR reduces tumor growth. Therefore, the multifunctional Cur-mSiO2@rGO can be a safe chemopreventive nanocarrier in tumor management and cancer photothermal therapy.

The Fisetin on Anti-senescence of Hepatocellular Carcinoma Cell

Liver cancer, as a global health issue, is of critical importance in termsal of treatment and prognosis. Quercetin, a flavonoid compound widely found in fruits, vegetables, and beverages, has become a research focus in recent years for its antioxidant, anti-inflammatory, anti-viral, and anti-tumor properties. Studies, especially on the inhibition of hepatocellular growth, the blocking of metastasis, and the regulation of cellular senescence mechanisms, have suggested that quercetin has potential therapeutic value. This article primarily reviews the progress of quercetin in the anti-senescence research of liver cancer cells. Studies have shown that quercetin can affect the proliferation, survival, and apoptosis of liver cancer cells through different molecular mechanisms. Firstly, quercetin can directly inhibit the growth of liver cancer cells by inducing cell cycle arrest at the G0/G1 or G2/M phases, and by upregulating pro-apoptotic proteins such as the caspase family and downregulating anti-apoptotic proteins like the Bcl-2 family, thereby promoting apoptosis of liver cancer cells. Secondly, quercetin can inhibit the invasion and metastasis of liver cancer cells, which may be related to its regulation of matrix metalloproteinase expression, significant in preventing distant metastases of liver cancer. Furthermore, quercetin can influence the biological behaviors of liver cancer cells by regulating related signaling pathways, such as PI3K/Akt, MAPKs, and NF-κB. In addition, quercetin exhibits a unique mechanism in regulating the senescence of liver cancer cells by promoting senescence through the activation of the p53/p21 and p16/Rb pathways. Finally, addressing the issue of quercetin's bioavailability, research has found that combining it with other drugs or improving the administration methods could enhance its therapeutic effect. In summary, as a multi-target natural compound, quercetin shows extensive potential in the treatment of liver cancer. However, its application in clinical therapy still requires further in vitro, in vivo experiments, and clinical studies to validate its effectiveness and safety.

CARM1 accelerates the growth of liver cancer cells by enhancing ARAF

CARM1 accelerates the growth of liver cancer cells by enhancing ARAF

Aristolochic acids-hijacked p53 promotes liver cancer cell growth by inhibiting ferroptosis.

Aristolochic acids (AAs) have been identified as a significant risk factor for hepatocellular carcinoma (HCC). Ferroptosis is a type of regulated cell death involved in the tumor development. In this study, we investigated the molecular mechanisms by which AAs enhanced the growth of HCC. By conducting bioinformatics and RNA-Seq analyses, we found that AAs were closely correlated with ferroptosis. The physical interaction between p53 and AAs in HepG2 cells was validated by bioinformatics analysis and SPR assays with the binding pocket sites containing Pro92, Arg174, Asp207, Phe212, and His214 of p53. Based on the binding pocket that interacts with AAs, we designed a mutant and performed RNA-Seq profiling. Interestingly, we found that the binding pocket was responsible for ferroptosis, GADD45A, NRF2, and SLC7A11. Functionally, the interaction disturbed the binding of p53 to the promoter of GADD45A or NRF2, attenuating the role of p53 inenhancing GADD45A and suppressing NRF2; the mutant did not exhibit the same effects. Consequently, this event down-regulated GADD45A and up-regulated NRF2, ultimately inhibiting ferroptosis, suggesting that AAs hijacked p53 to down-regulate GADD45A and up-regulate NRF2 in HepG2 cells. Thus, AAs treatment resulted in the inhibition of ferroptosis via the p53/GADD45A/NRF2/SLC7A11 axis, which led to the enhancement of tumor growth. In conclusion, AAs-hijacked p53 restrains ferroptosis through the GADD45A/NRF2/SLC7A11 axis to enhance tumor growth. Our findings provide an underlying mechanism by which AAs enhance HCC and new insights into p53 in liver cancer. Therapeutically, the oncogene NRF2 is a promising target for liver cancer.

Pomegranate’s silver bullet: Nature-powered nanoparticles deliver a one-two punch against cancer and antimicrobial resistance

Biogenic synthesis for producing silver nanoparticles (AgNPs) leverages natural extracts, offering an eco-friendly and biocompatible alternative to traditional chemical methods. Pomegranate (Punica granatum L. (PG)) extract, rich in phytochemicals, presents a valuable resource for this green synthesis approach. The primary objective is to harness the natural phytochemicals in PG extract to create stable, biocompatible AgNPs through a green synthesis method. Our study aims to explore the synthesis and characterization of AgNPs derived from PG using eco-friendly methods. In addition, this study investigates the antioxidant, antimicrobial, and anticancer properties of PG-AgNPs. The preparation of PG followed by the reduction of AgNO3 to PG-AgNPs, with subsequent characterization using UV–Vis spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). These techniques confirmed the successful synthesis of AgNPs with desirable size, shape, and stability. Further, antimicrobial properties, antioxidants, cytotoxicity assay, and ADME prediction were also assessed. The PG-AgNPs revealed no signs of aggregates, suggesting they could maintain their original shape. In addition, XRD analysis indicates that PG-AgNPs are highly pure. The zeta potentials of the biosynthesized PG-AgNPs were −17.8 ± 6.57 mV. AgNPs exhibited FTIR peaks associated with silanols, carboxylates, phosphonates, and siloxanes. The PG-AgNPs exhibited potent antibacterial activity against E. coli and Klebsiella pneumoniae, with higher efficacy than PG extract alone. Additionally, PG-AgNPs demonstrated significant antioxidant activity comparable to or exceeding that of ascorbic acid. The AgNPs effectively inhibited the growth of liver (HepG2), breast (MDA-MB-231), and cervical (HeLa) cancer cells in a dose-dependent manner while showing no toxicity to normal human cells (WI-38). The study concludes that biogenically synthesized PG-AgNPs possess potent anticancer and antimicrobial properties, highlighting their potential as therapeutic agents. The green synthesis method underscores the advantages of eco-friendly approaches in developing effective nanomaterials, paving the way for novel treatments in cancer therapy and combating antibiotic-resistant infections.

The 3,3'-dimethoxy-4,4'-dihydroxy-stilbene Triazole (STT) Inhibits Liver Cancer Cell Growth by Targeting Akt/mTOR Pathway.

The present study was aimed to investigate the proliferation inhibitory ability of 3,3'-dimethoxy-4,4'-dihydroxy-stilbene triazole (STT) on SNU449 and Huh7 cells. Moreover, the mechanism associated with the suppression of liver cancer cell proliferation by STT was also studied. The results revealed that STT suppresses proliferation of SNU449 and Huh7 cells to 28 and 21%, respectively treatment with 20 µM. The clonogenic survival of SNU449 and Huh7 cells was also significantly reduced after incubation with STT compared to the control cultures. In comparison to the control, STT treatment significantly decreased the invasive potential of SNU449 cells. Treatment with STT led to a prominent suppression in p62 and increase in LC3B protein expression in SNU449 cells compared to the control cells. The STT treatment dramatically decreased p-Akt and p-mTOR protein expression in SNU449 cells. Docking study revealed that STT interacts via traditional hydrogen bonding with the glutamine, phenylalanine, leucine, serine, arginine, aspartic acid, and lysine residues of Akt protein. In summary, the current study demonstrates that STT effectively suppresses the viability of SNU449 and Huh7 liver cancer cells. Moreover, STT treatment of the liver cancer cells also significantly reduces the clonogenic survival and invasive potential of SNU449 cells. Treatment of liver cancer cells with STT increases the expression of autophagic, targets anti-autophagic protein expression and down-regulates Akt/mTOR pathway to inhibit cancer growth and proliferation. Thus, STT exhibits prominent anticancer effect and needs to be investigated further as a potential candidate for the treatment of liver cancer.

Potential of β-D-glucan polysaccharide from Ophiocordyceps sinensis OS8 cultivated mycelium on anticancer activity via inducing liver cancer cell death apoptosis

The fungus Ophiocordyceps sinensis OS8 has been sourced from the wild variant of O. sinensis. In this study, the enriched β-D-glucan polysaccharide obtained from cultured mycelium of O. sinensis OS8 (OS8P) was examined for its potential to inhibit the growth of HepG2 liver cancer cells, and the IC50 was 511.79 µg/mL. The study of cellular migration was analyzed by scratch assay. It was found that the HepG2 cell migration was significantly inhibited (P < 0.05) in the OS8P-treated group. The cell apoptotic feature’s reliability was determined via DAPI staining to confirm the experimental results' reliability. The AO/PI double staining assay showed that HepG2 cells treated with OS8P (IC50) had dead cells with condensed and shrieking nuclei that are late apoptotic. Morphological alterations were meticulously examined using scanning electron microscopy. The specific morphological characteristics of apoptotic cells, notably cell lysis, were distinctly observable. The damaged cells demonstrated severe structural disruption, loss of original shape, blebs formation, and apoptotic bodies' appearance. These findings provide pharmacological proof that the OS8P extract could be potentially employed in the food and medical industries for anticancer applications.

The Prognostic Biomarker RAB7A Promotes Growth and Metastasis of Liver Cancer Cells by Regulating Glycolysis and YAP1 Activation.

Activating transcription factor 6 (ATF6) and its downstream genes are involved in progression of hepatocellular carcinoma (HCC). Herein, we demonstrated that sulfhydration of Ras-related protein Rab-7a (RAB7A) was regulated by ATF6. High expression of RAB7A indicated poor prognosis of HCC patients. RAB7A overexpression contributed to proliferation, colony formation, migration, and invasion of HepG2 and Hep3B cells. Furthermore, we found that RAB7A enhanced aerobic glycolysis in HepG2 cells, indicating a higher degree of tumor malignancy. Mechanistically, RAB7A suppressed Yes-associated protein 1 (YAP1) binding to 14-3-3 and conduced to YAP1 nuclear translocation and activation, promoting its downstream gene expression, thereby promoting growth and metastasis of liver cancer cells. In addition, knocking down RAB7A attenuated the progression of orthotopic liver tumors in mice. These findings illustrate the important role of RAB7A in regulating HCC progression. Thus, RAB7A may be a potential innovative target for HCC treatment.

Research progress of ferroptosis regulating lipid peroxidation and metabolism in occurrence and development of primary liver cancer.

As a highly aggressive tumor, the pathophysiological mechanism of primary liver cancer has attracted much attention. In recent years, factors such as ferroptosis regulation, lipid peroxidation and metabolic abnormalities have emerged in the study of liver cancer, providing a new perspective for understanding the development of liver cancer. Ferroptosis regulation, lipid peroxidation and metabolic abnormalities play important roles in the occurrence and development of liver cancer. The regulation of ferroptosis is involved in apoptosis and necrosis, affecting cell survival and death. Lipid peroxidation promotes oxidative damage and promotes the invasion of liver cancer cells. Metabolic abnormalities, especially the disorders of glucose and lipid metabolism, directly affect the proliferation and growth of liver cancer cells. Studies of ferroptosis regulation and lipid peroxidation may help to discover new therapeutic targets and improve therapeutic outcomes. The understanding of metabolic abnormalities can provide new ideas for the prevention of liver cancer, and reduce the risk of disease by adjusting the metabolic process. This review focuses on the key roles of ferroptosis regulation, lipid peroxidation and metabolic abnormalities in this process.

SHOC2 plays an oncogenic or tumor-suppressive role by differentially targeting the MAPK and mTORC1 signals in liver cancer

Abstract SHOC2 is a scaffold protein that activates the RAS-MAPK signal. Our recent study showed that SHOC2 is also a negative regulator of the mTORC1 signal in lung cancer cells. Whether and how SHOC2 differentially regulates the RAS-MAPK vs. the mTORC1 signals in liver cancer remains unknown. Here, we showed that SHOC2 is overexpressed in human liver cancer tissues, and SHOC2 overexpression promotes the growth and survival of liver cancer cells via activation of the RAS-MAPK signal, although the mTORC1 signal is inactivated. SHOC2 knockdown suppresses the growth of liver cancer cells mainly through inactivating the RAS-MAPK signal. Thus, in the cell culture models, SHOC2 regulation of growth is dependent of the RAS-MAPK but not the mTORC1 signal. Interestingly, in a mouse liver cancer model induced by diethylnitrosamine (DEN)-high-fat diet (HFD), hepatocyte-specific Shoc2 deletion inactivates the Ras-Mapk signal but has no effect in liver tumorigenesis. However, in the Pten loss-induced liver cancer model, Shoc2 deletion further activates mTorc1 without affecting the Ras-Mapk signal and promotes liver tumorigenesis. Collectively, it appears that SHOC2 could act as either an oncogene (via activating the MAPK signal) or a tumor suppressor (via inactivating the mTORC1 signal) in the manner dependent of the dominancy of the MAPK vs. mTORC1 signals.

Tryptophan fuels MYC-dependent liver tumorigenesis through indole 3-pyruvate synthesis

Cancer cells exhibit distinct metabolic activities and nutritional dependencies compared to normal cells. Thus, characterization of nutrient demands by individual tumor types may identify specific vulnerabilities that can be manipulated to target the destruction of cancer cells. We find that MYC-driven liver tumors rely on augmented tryptophan (Trp) uptake, yet Trp utilization to generate metabolites in the kynurenine (Kyn) pathway is reduced. Depriving MYC-driven tumors of Trp through a No-Trp diet not only prevents tumor growth but also restores the transcriptional profile of normal liver cells. Despite Trp starvation, protein synthesis remains unhindered in liver cancer cells. We define a crucial role for the Trp-derived metabolite indole 3-pyruvate (I3P) in liver tumor growth. I3P supplementation effectively restores the growth of liver cancer cells starved of Trp. These findings suggest that I3P is a potential therapeutic target in MYC-driven cancers. Developing methods to target this metabolite represents a potential avenue for liver cancer treatment.

Crithmum maritimum Extract Restores Lipid Homeostasis and Metabolic Profile of Liver Cancer Cells to a Normal Phenotype

Hepatocellular carcinoma (HCC) is an alarming epidemiological clinical problem worldwide. Pharmacological approaches currently available do not provide adequate responses due to poor effectiveness, high toxicity, and serious side effects. Our previous studies have shown that the wild edible plant Crithmum maritimum L. inhibits the growth of liver cancer cells and promotes liver cell differentiation by reducing lactic acid fermentation (Warburg effect). Here, we aimed to further characterise the effects of C. maritimum on lipid metabolism and markers of cellular metabolic health, such as AMP-activated protein kinase (AMPK), Sirtuin 1 (SIRT1), and Sirtuin 3 (SIRT3), as well as the insulin signalling pathway. To better mimic the biological spectrum of HCC, we employed four HCC cell lines with different degrees of tumorigenicity and lactic acid fermentation/Warburg phenotype. Lipid accumulation was assessed by Oil Red O (ORO) staining, while gene expression was measured by real-time quantitative PCR (RT-qPCR). The activation of AMPK and insulin signalling pathways was determined by Western blotting. Results indicate that C. maritimum prevents lipid accumulation, downregulates lipid and cholesterol biosynthesis, and modulates markers of metabolic health, such as AMPK, SIRT1 and SIRT3. This modulation is different amongst HCC cell lines, revealing an important functional versatility of C. maritimum. Taken together, our findings corroborate the importance of C. maritimum as a valuable nutraceutical, reinforcing its role for the improvement of metabolic health.

Inhibition of Liver Cancer Cell Viability by Triazole through Up-regulation of p38 Phosphorylation and Targeting the Activation of p-ERK1/2 and Akt Protein Expression.

The present study was aimed to explore the effect of triazole on growth and viability of liver cancer cells. Cell growth was examined using the MTT test and expression of several proteins was assessed by western blotting assay. The Matrigel-coated Transwell assay was employed to examine the infiltration of cells. The data from MTT assay showed that MHCC97H and H4TG liver cancer cell viability was inhibited by triazole in a concentration-dependent manner. After treatment with 0.5, 1.0, 2.0, 4, 8, and 16 µM doses of triazole, the rate of H4TG cell viability was decreased to 96, 73, 58, 39, 29, and 28%, respectively. Treatment of MHCC97H cells with 0.5, 1.0, 2.0, 4, 8, and 16 µM doses of triazole resulted in a reduction in cell viability to 94, 70, 53, 35, 22, and 21%, respectively. Triazole treatment also led to a significant reduction in MHCC97H cell invasiveness compared to the control cells. In MHCC97H cells treated with triazole, there was a noticeable decrease in the levels of p-ERK1/2, and p-Akt protein expression. Treatment of MHCC97H cells with triazole resulted in a prominent increase in p-p38 level. In summary, triazole inhibits growth and viability of liver cancer cells through targeting the activation of p-ERK1/2 and Akt proteins. Therefore, triazole may be investigated further as a therapeutic agent for the treatment of liver cancer.

Abstract 7197: Elacridar sensitizes liver cancer to the PKC-inhibitor ICA-1S

Abstract Liver cancer is a disease in which cancer cells form in the tissues of the liver. Globally, liver cancer is one of the most frequent fatal malignancies; in the United States, liver cancer is the sixth leading cause of cancer death. Generally, there is a poor prognosis for liver cancer. Chemotherapy and immunotherapy are the best available treatment options. This research aims to provide an alternative treatment option with drugs that specifically attack liver cancer cells. Protein Kinase C (PKCs) are signaling molecules that have been reported to be associated with multiple cellular processes and cancer. PKC activity reportedly is elevated in colorectal cancer cells and metastasizes to the liver. 5-amino-1-((1R,2S,3S,4R)-2,3-dihydroxy-4-methylcyclopentyl)-1H-imidazole-4-carboxamide (ICA-1S) is a PKC-ι specific inhibitor we have previously shown to be effective against some cancer cell lines by decreasing cell growth and inducing apoptosis. In this study, we investigated the effectiveness of ICA-1S against liver cancer cell lines; HEPG2 and PLC5/PRF/5. An equal number of HEPG2 and PLC5/PRF/5 cell lines were seeded into 6 well plates, treated with different concentrations (0.1µM, 0.5µM, 1µM, 5µM, and 10µM) of ICA-1s every 24 hours for three days after which the media was discarded, and cells were counted. The percentage inhibition of ICA-1S on HEPG2 and PLC5/PRF/5 cell lines was calculated. To investigate the resistance inflection, the proliferation assay was also conducted with a combination of an efflux pump inhibitor, Elacridar(10µM), and different concentrations (0.1µM to 10µM) of ICA-1S. The expression of PKC-ι was investigated across all treatments. The dose-response result for treatment with ICA-1S for both HEPG2 and PLC5/PRF/5 cell lines showed a resistance inflection point at 0.5µM and 1µM, where the proliferation rate of the cells declined by 50% and 79% for HEPG2, and 41% and 54% for PLC5/PRF/5 respectively; beyond these points, the cell proliferation rate significantly rises. The results of dose-response for treatment with a combination of ICA-1S and Elacridar showed a continuous decline in cell proliferation rate up to 78% and 84% for HEPG2 and PLC5/PRF/5 respectively as the concentrations of ICA-1S increased from 0.1µM to 10µM. The PKC-ι level was downregulated in the combination treatment. We have shown in this study that liver cancer cells exhibit resistance to PKC inhibitor, ICA-1S, at high concentrations. This resistance inflection may be due to several factors including the activation of drug-efflux proteins. Thus, we proved that the introduction of an efflux pump inhibitor, Elacridar, sensitizes the liver cancer cells to ICA-1S, offsets the resistance of the liver cancer cells to ICA-1S at high concentrations, and enables ICA-1S to be effective in downregulating PKC-ι and decreasing cell growth of liver cancer cells. Citation Format: Abigail Oluwafisayo Olatunji, Mahfuza Marzan, Mildred Acevedo-Duncan. Elacridar sensitizes liver cancer to the PKC-inhibitor ICA-1S [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7197.