Hepatic Carcinoma Cells Research Articles

Prospective Study of Isolated Recurrent Tumor Re-irradiation With Carbon-Ion Beams.

Purpose: To perform a prospective study to evaluate the efficacy and safety of isolated recurrent tumor re-irradiation with carbon-ion radiotherapy (RT).Methods and Materials: The inclusion criteria were clinically proven recurrent tumors, measurable by computed tomography or magnetic resonance imaging, patients ≥ 16 years old, performance status scores between 0 and 2, isolated tumor at a previously irradiated site, and a life expectancy > 6 months. The exclusion criteria were tumor invasion into the gastrointestinal tract or a major blood vessel, uncontrolled infection, early recurrence (<3 months), and severe concomitant diseases. The primary end-point was the local control rate, the secondary end-points including the overall survival rate, and adverse events.Results: Between December 2013 and March 2016, 22 patients were enrolled in this prospective study. All patients were re-irradiated with carbon-ion RT with radical intent. Five patients had rectal cancer, 4 had sarcoma, 4 had lung cancer, 3 had hepatic cell carcinoma, and 6 had other tumors. The median follow-up time was 26 months. Eight patients developed local recurrence, and the 1- and 2-year local control rates were 71 and 60%, respectively. Eight patients died of their cancers and 2 died of other diseases. The 1- and 2-year overall survival rates were 76 and 67%, respectively. There were no grade 2 or higher acute adverse events and 4 patients (18%) developed grade 3 late adverse events. The group with the longer interval (>16 months) between the first RT and re-irradiation had significantly better outcomes than the shorter interval group (≤ 16 months).Conclusions: Re-irradiation, using carbon-ion RT with radical intent, had favorable local control and overall survival rates without severe toxicities for selected patients. Re-irradiation has the potential to improve clinical outcomes for isolated, local, recurrent tumors; further investigations are required to confirm the therapeutic efficacy.

PZR promotes metastasis of colorectal cancer through increasing FAK and Src phosphorylation.

Metastasis is the main cause of death in patients with colorectal cancer (CRC), but the molecular mechanism is not yet fully understood. Previous studies have shown that P zero-related protein (PZR), a member of the immunoglobulin family, can promote fibronectin-dependent migration of mouse embryonic fibroblasts as well as invasion and metastasis of hepatic carcinoma cells. However, the role of PZR in CRC remains unclear. In this study, we determined the ectopic expression of PZR in CRC tissues, and results showed that PZR expression was increased not only in tumors with higher pathological stage, but also in tumors with distant metastasis. Through PZR-knockdown and overexpression in CRC cell lines, we found that the expression of PZR had significant effect on the invasion and migration of CRC cells as well as the phosphorylation of pro-metastasis proteins including focal adhesion kinase (FAK) and Src. Taken together, this study indicates that PZR may promote the invasion and migration of CRC cells through increasing the phosphorylation of FAK and Src, which provides a new theoretical basis and a possible marker for the diagnosis or prognosis of CRC metastasis.

Dextrose modified bilosomes for peroral delivery: improved therapeutic potential and stability of silymarin in diethylnitrosamine-induced hepatic carcinoma in rats

Hepatic carcinoma (HC) is one of the most prevalent cancers, ranked as the second most common cause of cancer-related deaths worldwide. Silymarin (SYL) has been reported for its anticarcinogenic activity against various types of cancer such as prostate, breast, ovary, colon, lung, bladder and liver. Due to poor solubility and low bioavailability SYL lacks satisfactory therapeutic value thus designing a suitable and effective delivery system of SYL can led to improved therapeutic potential. The present study was aimed to develop SYL-loaded dextrose (DEX) modified bilosomes for targeted delivery to HC cells. The DEX-modified bilosomes were prepared through thin-film hydration method and optimized employing Box Behnken design. The bilosomes were evaluated for percent entrapment, drug loading, in vitro release and cytotoxicity on Hep-G2 cells. The optimized DEX-SYL-BL exhibited a particle size of 219.3 ± 2.99 nm, percent entrapment of 62.32 ± 4.23%, drug loading of 34.56 ± 1.23% and 84.96 ± 2.76% drug release respectively over a period of 24 hr. The stability of bilosomes was ascertained in simulated gastric and intestinal fluids. Cytotoxicity studies revealed greater performance of DEX-SYL-BL in terms of reduced viability in Hep-G2 cell lines when compared with pure SYL and SYL-BL. Further DEX-modified bilosomes were evaluated in vivo for their therapeutic efficacy in DEN-induced (Diethylnitrosamine) hepatic carcinoma in animal model. The DEX-SYL-BL displayed higher therapeutic potential as revealed from enhanced survival and reduced tumour burden in animals. DEX-SYL-BL also displayed significant restoration of altered oxidative markers and SGOT, SGPT levels towards normal value when compared with pure SYL.

In vitro study of BSA gel/polyelectrolite complexes core shell microcapsules encapsulating doxorubicin for antitumoral targeted treatment

The aim of this study is to develop core shell microcapsules of bovine serum albumin (BSA) gel with a complex polyelectrolite multilayer shell of natural polysaccharides with opposite charges, pectin (P), chitosan (Chi), and hyaluronic acid (HA) respectively, encapsulating Doxorubicin (Dox) as a carrier for targeted anti-tumoral treatment of hepatic cell carcinoma (HCC). A sacrificial CaCO3 template method was used in order to obtain microcapsules with a BSA gel core and a layer-by-layer (Lbl) deposition technique of polyelectrolite complexes formed between P/Chi in the inner layers and HA/Chi in the outer shell layers. The preformed microcapsules, BSA gel/P/Chi/HA, noted as ms, have been applied for Dox encapsulation (ms-Dox). Dox encapsulation and release in different pH media were studied in order to elucidate the interactions between pH dependently charged species involved in the Dox loading/releasing processes. The structure characterization of ms/ms-Dox was evaluated by FTIR and UV-Vis spectroscopy, X-ray diffraction, thermal analy sis, optical microscopy, confocal laser scanning microscopy, and scanning electron microscopy. The in vitro study for citotoxicity assessment on normal and tumoral cells of both ms and ms-Dox was performed using mesenchymal stem cells (MSCs) and Hep2G HCC cell lines. Results of physical-chemical analyses confirm the successful encapsulation of Dox in ms, and the in vitro biological study recommends ms-Dox as a candidate for future in vivo research as a targeted anti-tumoral treatment modality applications.

Protein arginine methyltransferase 1 promotes epithelial-mesenchymal transition via TGF-β1/Smad pathway in hepatic carcinoma cells.

Protein arginine methyltransferase 1 (PRMT1) is dysregulated in a number of human cancers. Herein, we report that PRMT1 expression is directly associated with epithelial-mesenchymal transition (EMT) in hepatic carcinoma cells. Firstly, we find that PRMT1 expression is higher in hepatic carcinoma tissues than that in normal liver tissues at both mRNA and protein levels, and higher expression of PRMT1 correlates with poor survival in liver tumors. The data in vitro reveals that PRMT1 knockdown inhibits the abilities of proliferation, migration and invasion, while PRMT1 overexpression promotes the above behaviors in hepatic carcinoma cells. Further studies indicate that PRMT1 knockdown remarkably decreases the expression of mesenchymal markers including Vimentin, Snail and N-cadherin, and upregulates expression of epithelial markers E-cadherin. Conversely, PRMT1 overexpression results in the opposite effects. Additionally, we identified that PRMT1 knockdown resulted in downregulation of TGF-β1, p-Smad2 and p-Smad3, while PRMT1 overexpression activated TGF-β1, p-Smad2 and p-Smad3. These findings suggest that PRMT1 promotes EMT in hepatic carcinoma cells probably via TGF-β1/Smad pathway, and might represent a novel anti-liver cancer strategy.

Cancer mortality among indigenous population in Acre State, Brazil

The study aimed to estimate cancer mortality among indigenous peoples in Acre State, Brazil. This was a descriptive observational study based on the nominal bank of the Brazilian Mortality Information System for the period from January 1st, 2000, to December 31st, 2012. The study analyzed the distribution death frequencies by sex and age. Standardized mortality ratio (SMR) was calculated taking Goiânia (Goiás State), Acre State, and the North Region of Brazil as the references. A total of 81 deaths were identified, the majority in men (59.3%) and in individuals over 70 years of age. The five main sites in men were stomach, liver, colon and rectum, leukemia, and prostate. The five main sites in women were uterine cervix, stomach, liver, leukemia, and uterus. In indigenous men there was an excess of deaths from stomach cancer compared to the populations of Goiânia (SMR = 2.72; 2.58-2.87), Acre State (SMR = 2.05; 1.94-2.16) and North region (SMR = 3.10; 2.93-3.27). The same was observed for deaths from hepatic cell carcinomas referenced against Goiânia (SMR = 3.89; 3.66-4.14), Acre State (SMR = 1.79; 1.68-1.91), and the North of Brazil (SMR = 4.04; 3.77-4.30). Among indigenous women, there was an excess of cervical cancer in comparison to Goiânia (SMR = 4.67; 4.41-4.93), Acre State (SMR = 2.12; 2.00-2.24), and the North (SMR = 2.60; 2.45-2.75). The estimates show that preventable neoplasms such as cervical cancer and those linked to underdevelopment, such as stomach and liver cancer, account for 49.4% of deaths among indigenous peoples. Compared to the reference population, mortality from liver, stomach, and colorectal cancer and leukemias was more than twice as high in indigenous men; among indigenous women, cervical, stomach, and liver cancer and leukemias were 30% higher.

Rosmarinic Acid Combined with Adriamycin Induces Apoptosis by Triggering Mitochondria-Mediated Signaling Pathway in HepG2 and Bel-7402 Cells.

BackgroundHepatic carcinoma is the third leading cause of cancer-related deaths. This study aimed to evaluate the anti-tumor effects of rosmarinic acid (RosA) combined with Adriamycin (ADM) on proliferation and apoptosis of hepatic carcinoma cell lines.Material/MethodsHuman HepG2 and Bel-7402 cells were treated with RosA and ADM and divided into HepG2 or Bel-7402, 25 μg/ml, 50 μg/m, and 100 μg/ml RosA+0.4 μg/ml ADM groups, respectively. The Cell Counting Kit-8 (CCK-8) assay was used to evaluate cell viability. Immunohistochemistry assay was used to examine B cell lymphoma-2 (Bcl-2) and Bcl-2-associated X (Bax) expression. Cell cycle analysis was used to detect cell cycle distribution. Flow cytometry and terminal deoxynucleotidyl transferase-mediated d-UTP nick-end labeling (TUNEL) assay were utilized to evaluate apoptosis.ResultsRosA combined with ADM damaged cell morphology and decreased cell viability, and significantly decreased S-phase cell numbers compared to the HepG2 or Bel-7402 group (p<0.05). Apoptosis rates in the RosA combined with ADM group were significantly increased compared to the HepG2 or Bel-7402 group (p<0.05). TUNEL assay showed that RosA combined with ADM significantly induced DNA damage (TUNEL-positive staining) in the HepG2 and Bel-7402 groups (p<0.05). RosA combined with ADM significantly reduced Bcl-2 expression in HepG2 or Bel-7402 cells (p<0.05). RosA combined with ADM significantly increased Bax expression in HepG2 and Bel-7402 cells (p<0.05). Cell viability, apoptosis, cell cycle, and Bcl-2 and Bax expression were changed with increased concentrations of RosA.ConclusionsRosA combined with ADM damaged tumor cell morphologies, decreased cell viability, and induced apoptosis of HepG2 and Bel-7402 by triggering the mitochondria-mediated signaling pathway.

Ozone detoxification of patulin in aqueous solution and cytotoxic evaluation using human hepatic carcinoma cells

Patulin often contaminates fruits and fruit-base products, which seriously threats the health of consumers. In this study, ozone was used to degrade patulin in aqueous solution, and investigated the cytotoxicity of patulin after ozone detoxification on human hepatic carcinoma cells (HepG2) using MTT assay and apoptosis assay. Patulin was rapidly degraded from 24.59 mg/L to 9.85 mg/L within 180 s by 10.60 mg/L of ozone at a flow rate of 90 mL/min, and reduced by 59.94%. The half maximal inhibitory concentration (IC50) of patulin on HepG2 cells was 9.32 μmol/L after 24 h of exposure, and it showed a dose dependent effect. After 90 s of ozone detoxification, the cell viability of HepG2 cells obviously increased from 42.31% to 93.96%, and the total apoptotic cells significantly reduced from 22.24% to 11.18% after 180 s of ozone treatment. The results clearly show the great potential of ozone in degrading patulin in liquid foods.

Construction of the pEGFP-N1-p53/MAR Vector and Its Effect on HEP3B Cell Morphology

Cancer always presents a big problem that endangers human health. In recent years, the use of gene therapy in cancer research has significantly increased. This study aimed to construct a non-viral, wild-type, recombinant eukaryotic expression vector, pEGFP-N1-p53/MAR and verify its mechanism of action in cancer cells in vitro. This investigation provides a novel strategy for p53 gene therapy via regulation of the matrix attachment region (MAR), potentially laying a foundation for the establishment of an anticancer protein bioreactor. The p53 gene was cloned from human peripheral blood and the MAR gene was amplified from chicken liver tissue. The recombinant eukaryotic expression vector pEGFP-N1-p53/MAR was constructed using an E. coli self-replication system. LipofectamineTM 2000 was used as the transfection agent to deliver the plasmid into the human hepatic carcinoma (HEP3B) cell line. We divided the groups as follows: negative control cells without plasmid transfection, vehicle control cells transfected with the PEGFP-N1 vector, and experimental cells transfected with the pEGFP-N1-p53/MAR vector. Cells in each well of the vehicle control and experimental groups were transfected with 1.6 μg of plasmid and 3 μL of liposome. The cellular morphology of each group was analysed using green fluorescence microscopy at 12, 24, 36 and 48 h. Then, statistical analysis of the apoptosis rates among the three groups was performed using SPSS. The ultrastructures of the cells were observed via transmission electron microscopy after transfection for 24 h. Morphological analysis showed that the cells of the experimental group were shrunken and reduced in size and their intercellular connections had disappeared. Additionally, the apoptosis rate in the experimental group was significantly higher than that in the control groups and the cellular microstructure showed that heterochromatin and apoptotic bodies were found in the experimental group. In conclusion, compared with the control groups, the pEGFP-N1-p53/MAR plasmid can effectively promote Hep3B cell apoptosis in vitro.

Chrysin promotes SMMC-7721 cell apoptosis by regulating MAPKs signaling pathway

To study the effect of chrysin in inducing apoptosis of human hepatic carcinoma cells and explore the possible mechanism. Human hepatic carcinoma SMMC-7721 cells treated with DMSO or chrysin at different concentrations (5-200 μg/mL) were examined for changes in the cell proliferation using CCK-8 assay. The morphological changes of SMMC-7721 cells were observed in response to treatment with 5, 10, or 20 μg/mL chrysin, and the changes in the cell nuclei were observed using DAPI nuclear staining. Annexin Ⅴ-FITC/PI flow cytometry was used to determine the cell apoptosis rate. The changes in the apoptosis-related proteins (PARP and caspase-3) and MAPKs signal pathway were detected with Western blotting. Chrysin treatment obviously suppressed the proliferation of SMMC-7721 cells in a dose-dependent manner below the concentration of 60 μg/mL. Chrysin (20 μg/mL) also caused significantly increased cell apoptosis and significant cleavage of PARP and caspase-3. Chrysin significantly activated MAPKs signaling pathway in a time-and dose-dependent manner, with the peak activation level occurring at 15 min. Pretreatment of the cells with specific inhibitors of the MAPKs pathway obviously inhibited the effect of chrysin in inducing cell apoptosis. Chrysin inhibits the proliferation and promotes apoptosis of SMMC-7721 cells by regulating the activation of MAPKs signaling.

The AT1/Raf/ERK1/2 signaling pathway is involved in Angiotensin II-enhanced proliferation of hepatic carcinoma cells.

Angiotensin II (Ang II) has been strongly associated with biological behavior in human malignant tumors; nevertheless, its function in hepatic carcinoma growth and progression is still not well understood. This study investigates the effect and mechanism of Ang II on the HepG2 and Hep3B hepatic carcinoma cell lines in vitro. The effect of Ang II on HepG2 and Hep3B cell viability was examined by cell counting kit-8 assay (CCK-8). Quantitative real-time PCR and Western blot analysis detected the expression of angiotensin type 1 and type 2 receptors (AT1 and AT2), total extra-cellular signal-regulated kinases 1/2 (ERK1/2), phospho-ERK1/2 (p-ERK1/2) and Bcl-2 and c-Myc. Ang II significantly promoted HepG2 cell proliferation by affecting AT1 and AT2 expression and induced ERK1/2 pathway activation. This was reversed by treating HepG2 and Hep3B cells with AT1 blockers; candesartan, Raf inhibitor sorafenib, and ERK1/2 inhibitor PD98059. Ang II also up-regulated the expression of Bcl-2 and c-Myc in HepG2 cells, and our results suggest that Ang II has a positive role in HepG2 and Hep3B cell proliferation through the AT1/Raf/ERK1/2 signaling pathway.

Silver-doped graphene oxide nanocomposite triggers cytotoxicity and apoptosis in human hepatic normal and carcinoma cells.

IntroductionGraphene oxide nanoparticles have been widely used in industry and biomedical fields due to their unique physicochemical properties. However, comparative cytotoxicity of silver-doped reduced graphene oxide (rGO–Ag) nanoparticles on normal and cancerous liver cells has not been well studied yet.Materials and methodsThis study aimed at determining the toxic potential of rGO–Ag nanocomposite on human liver normal (CHANG) and cancer (HepG2) cells. The rGO–Ag nanocomposite was characterized by using different advanced instruments, namely, dynamic light scattering, scanning electron microscope, and transmission electron microscope.ResultsThe rGO–Ag nanocomposite reduced cell viability and impaired cell membrane integrity of CHANG and HepG2 cells in a dose-dependent manner. Additionally, it induced reactive oxygen species generation and reduced mitochondrial membrane potential in both cells in a dose-dependent manner. Moreover, the activity of oxidative enzymes such as lipid peroxide, superoxide dismutase, and catalase were increased and glutathione was reduced in both cells exposed to rGO–Ag nanocomposite. Pretreatment with N-acetylcysteine inhibited cytotoxicity and reactive oxygen species generation in CHANG and HepG2 cells exposed to rGO–Ag nanocomposite (50 µg/mL). DNA damage was determined by Comet assay and maximum DNA damage occurred at rGO–Ag nanocomposite (25 µg/mL) for 24 h. It is also valuable to inform that HepG2 cells appear to be slightly more susceptible to rGO–Ag nanocomposite exposure than CHANG cells.ConclusionThis result provides a basic comparative toxic effect of rGO–Ag nanocomposite on hepatic normal and cancerous liver cells.

Anti-inflammatory and cytotoxic withanolides from Physalis minima

Six undescribed withanolides were isolated and characterized during the investigation of anti-inflammatory and cytotoxic constituents from the whole plants of Physalis minima L. Their structures were elucidated by extensive spectroscopic analyses (IR, UV, HR-ESI-MS, 1D-NMR, and 2D-NMR), and their anti-inflammatory and cytotoxic activities were evaluated in vitro. All the compounds exhibited anti-inflammatory ability via inhibiting the production of nitric oxide (NO) in lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells. Moderate cytotoxic activities against A549 lung adenocarcinoma cells, SMMC-7721 hepatic carcinoma cells and MCF-7 breast cancer cells with IC50 values in the range of 40.01–82.17 μM were observed for these withanolides.

Cancer Cell-Intrinsic PD-1 and Implications in Combinatorial Immunotherapy.

Programmed death 1 (PD-1) and its two natural ligands PD-L1 and PD-L2 are responsible for delivering inhibitory signals that regulate the balance between T cell activation, tolerance, and immunopathology. In previous studies, PD-1 was found only expressed on the surface of immune cells, such as T cells and B cells while PD-1’s ligands PD-L1 and PD-L2 were found expressed in some tumor cells. However, recent studies revealed intrinsic expression of PD-1 in melanoma and some other cancers. In melanoma cells, PD-1 can be activated by its ligand PD-L1 expressed by tumor cells, modulating downstream mammalian target of rapamycin signaling and promoting tumor growth independent of adaptive immunity. In addition to melanoma, PD-1 was also detected in liver cancer cells as well as in non-small lung cancer cells. Unlike its oncogenic functions in melanoma and hepatic carcinoma cells, PD-1 seemed to play a distinct role in lung cancer, as blockade of PD-1 instead promoted tumor cells proliferation. Tumor-intrinsic PD-1 expression seems to be widespread in many tumor types, according to our reanalysis on cancer transcriptomic and proteomic data. The multifaceted roles of PD-1 in tumor cells beyond immune checkpoint signaling may explain the differential therapeutic effects of anti-PD-1 and anti-PD-L1 drugs and provide crucial information when developing combinatorial approaches to enhance antitumor immunity.

Mechanisms of G0/G1 phase arrest induced by longevity-assurance homologue 2 in Hepa1-6 hepatoma cells

Objective To investigate the effects of longevity-assurance homologue gene 2 (Lass2) on the cell cycle ofmouse hepatic carcinoma cell line Hepa1-6 and the related mechanism. Methods Lass2 recombinant eukaryotic expression vector has been successfully constructed in the early stage, and were transfected in Hepa1-6 hepatoma cell line 48 h by liposome 2000. The cell cycle of each group (negative control, empty load, experimental group) was detected by flow cytometry (PI). The relative expression levels of Lass2, Cyclin D1, nuclear factor-κB (NF-κB) p65 mRNA and protein were detected by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blotting. The phosphorylation of NF-κB p65 protein were detected by Western blotting; In order to further verify and clarify the mechanism, the C57BL/6J mouse hepatic carcinoma in situ model was constructed by injecting Hepa1-6 hepatic carcinoma cell suspension directly into the liver parenchyma of C57BL/6J mice. The naked plasmid was injected into the tail vein separately by hydrodynamics, and the total RNA and total protein in liver tissue of each group were extracted after transfection. The relative expression levels of Lass2, Cyclin D1, NF-κB p65 mRNA and protein in mice liver were detected by FQ-PCR and Western blotting. The phosphorylation of NF-κB p65 protein were detected by Western blotting. Results The relative expression levels of Lass2 mRNA and protein in Hepa1-6 hepatoma cells were significantly higher than those in negative control group and blank control group (F=1 486.895, all P=0.000; protein: F=55.964, all P=0.000), and the percentage of cell cycle G0/G1 phase increased significantly (F=239.867, all P=0.000), significantly decreased in G2/M phase and S phase (FG2/M=13.462, PG2/M=0.003, 0.005; FS=46.207, all P=0.000), at the same time, the levels of NF-κB p65, CyclinD1 mRNA and protein in the experimental group were decreased (mRNA: FNF-κB p65=29.993, FCyclin D1=113.260, all P=0.000; protein: FNF-κB p65=17.679, P=0.003, 0.002; FCyclin D1=176.357, all P=0.000), the phosphorylation of NF-κB p65 were significantly downregulated (FNF-κB p-p65=33.817, P=0.000). Conclusion Lass2 gene blocks the cell cycle of Hepa1-6 cells in G0/G1 phase, and its mechanism may be related to inhibits NF-κB p65 activation and expression and down-regulation of Cyclin D1 transcription and protein expression. Key words: Longevity-assurance homologue gene 2; Hepatocellular carcinoma; Cell cycle; Nuclear factor-κB p65; Cyclin D1

Targeted and synergistic therapy for hepatocellular carcinoma: monosaccharide modified lipid nanoparticles for the co-delivery of doxorubicin and sorafenib.

PurposeTargeted hepatocellular carcinoma therapy was carried out to improve the efficacy of liver cancer treatment. The purpose of this study was to design an N-acetylgalactosamine (NAcGal) modified and pH sensitive doxorubicin (DOX) prodrug (NAcGal-DOX) for the construction of lipid nanoparticles (LNPs).MethodsNAcGal-DOX and sorafenib (SOR) co-loaded LNPs were designed and the synergistic effects were evaluated on human hepatic carcinoma (HepG2) cells in vitro and anti-hepatic carcinoma mice model in vivo.ResultsCellular uptake efficiency of NAcGal modified LNPs was significantly higher than unmodified LNPs. NAcGal modified LNPs showed the most significant inhibition effect among all the samples tested. The results revealed that the LNPs system achieved significant synergistic effects, best tumor inhibition ability and the lowest systemic toxicity.ConclusionThese results proved that the NAcGal conjugated and pH sensitive co-delivery nano-system could be a promising strategy for treatment of hepatocellular carcinoma.

Enhanced cytotoxic and apoptotic potential in hepatic carcinoma cells of chitosan nanoparticles loaded with ginsenoside compound K

Ginsenoside compound K (CK) has been shown to exhibit anticancer properties. In this study, chitosan nanoparticles loaded with ginsenoside compound K (CK-NPs) were prepared as a delivery system using a self-assembly technique with amphipathic deoxycholic acid-O carboxymethyl chitosan as the carrier, which improved the water solubility of CK. By evaluating drug loading, entrapment efficiency, and in vitro release behavior, the feasibility of CK-NPs as a drug carrier nanoparticle for the treatment of human hepatic carcinoma cells (HepG2) was investigated. Result revealed that CK and CK-NPs showed a dose-dependent inhibitory effect on HepG2 cells with IC50 values of 23.33 and 16.58 μg/mL, respectively. Furthermore, fluorescence imaging demonstrated that CK-NPs promoted cellular uptake in vitro. Therefore, all results indicated that CK-NPs might be a novel drug delivery system to improve the solubility and enhance the cytotoxic and apoptotic potentials of CK for effective liver cancer chemotherapy.

N-phenylsulfonyl-3,5-bis(arylidene)-4-piperidone derivatives as activation NF-κB inhibitors in hepatic carcinoma cell lines

Ten novel symmetric 3,5-bis(arylidene)-4-piperidone derivatives (BAPs, 1–10) and fourteen dissymmetric BAPs (11–24) were synthesized and evaluated the cytotoxicity. All of the compounds have been screened for their anti-inflammatory activity characterized by evaluating their inhibitory effects on LPS-induced IL-6, TNF-α secretion. Among them, BAP 23 exhibits both the highest anti-inflammatory and anti-cancer properties. Western blot analysis showed that BAP 23 markedly reduced the levels of Bcl-2 but increased the levels of cleaved caspase-3and Bax. Moreover, BAP 23 prominently inhibited LPS-induced activation of NF-κB in RAW264.7 cells as well as TNF-α-induced activation of NF-κB in HepG2 cells by blocking phosphorylation of inhibitor IκBα and p65. Consistent with these results, we found that BAP 23 prevented the nuclear translocation of NF-κB induced by LPS or TNF-α. BAP 23 could reasonably bind to the active site of Bcl-2 protein and p65 which is proved by molecular docking modes. These data indicate that BAP 23 is a more potent inhibitor of NF-κB activity which exhibites both anti-inflammatory and anticancer activities.

Anesthetic effects of isoflurane and the molecular mechanism underlying isoflurane‑inhibited aggressiveness of hepatic carcinoma.

Anesthesia is produced by drugs or other methods, and refers to the attenuation of pain via reversible suppression of neuronal transmission in the central and peripheral nervous systems, during surgery. Clinical investigations have indicated that the anesthetic action of isoflurane is efficient to alleviate pain during tumor resection clinical trials. In addition, it has been reported that isoflurane can induce caspase-3 activation and is associated with apoptosis of tumor cells. The present study investigated the anesthetic effects and molecular mechanisms underlying isoflurane-induced apoptosis in patients with hepatic carcinoma. Furthermore, the pain of patients with hepatic carcinoma was evaluated during the perioperative period according to the pain index. The apoptotic rate of hepatic carcinoma cells was analyzed in tumor tissues using TUNEL assay. The expression levels of apoptosis-associated proteins were detected in liver cancer cells following anesthesia in patients. Phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) and nuclear factor (NF)-κB signaling pathways were also analyzed in liver cancer cells following treatment with isoflurane. The results demonstrated that isoflurane inhibited growth and decreased viability of liver cancer cells in vitro and in vivo. In addition, the apoptotic rate was increased in cells obtained from isoflurane-treated patients. The results also demonstrated that isoflurane upregulated the expression levels of proapoptotic genes and downregulated anti-apoptotic mRNA expression. In addition, a molecular mechanism analysis indicated that isoflurane inhibited PI3K and AKT expression in liver cancer cells. Isoflurane also induced caspase-3 activation in liver cancer cells. Furthermore, isoflurane treatment attenuated NF-κB activity and inhibited migration and invasion of liver cancer cells. In conclusion, these findings indicated that isoflurane treatment efficiently attenuated surgical pain and inhibited tumor aggressiveness via regulation of NF-κB activity and the PI3K/AKT signaling pathway, thus suggesting that isoflurane is an efficient anesthetic drug that induces pain remission and promotes apoptosis of liver cancer cells.

ISOLATION OF BIOACTIVE COMPOUNDS FROM CENTAUREA AEGYPTIACA

Objective: In a previous study, Centaurea aegyptiaca ethanol and ethyl acetate extracts showed potent cytotoxic effects against laryngeal (HEP2) and hepatic (HEPG2) carcinoma cell lines. Additionally, two novel compounds were isolated and identified. The aim of this study is to continue isolating and identifying another compound (s) that may, also, be responsible for this potent biological activity.Methods: C. aegyptiaca dried aerial parts were extracted with ethanol and ethyl acetate. Both extracts were chromatographed separately to afford seven guaianolides that were identified using different spectroscopic methods. Moreover, compounds 1-7 were evaluated for their cytotoxicity (IC50, µM) against HEP2 and HEPG2 cells in comparison to the normal fibroblasts (BHK) using sulforhodamine B assay. Doxorubicin was used as a positive control.Results: Seven sesquiterpene lactones, centaurepensin, also known as chlorohyssopifolin A (1), 8α-hydroxy-11α, 13-dihydrozaluzanin C (2), chlorohyssopifolin B (3), desacylcynaropicrin (4), chlorohyssopifolin C, acroptilin (5), subluteolide (6), and solstitiolide (7) were isolated from C. aegyptiaca extracts and identified. This is the first report on the occurrence of 2, 4, 5 and 6 in C. aegyptiaca. Compounds 1-4 and 6 exhibited selective cytotoxic effects against HEP2 and HEPG2 cells. However, compounds 1 and 7 showed the highest activities against HEP2 with IC50 values of 10.6±0.02 and 10.9±0.03 µM, respectively. Moreover, compound 3 was the most potent one against HEPG2 cells with IC50value of 13.8±0.05 µM.Conclusions: Chemical investigation of C. aegyptiaca ethanol and ethyl acetate extracts led to the isolation and identification of seven guaianolides. These compounds exhibited good cytotoxic activities against HEP2 and HEPG2 cell lines.