Growth Inhibition Of HepG2 Cells Research Articles

Long noncoding RNAs HAND2-AS1 ultrasound microbubbles suppress hepatocellular carcinoma progression by regulating the miR-873-5p/tissue inhibitor of matrix metalloproteinase-2 axis.

Increasing data indicated that long noncoding RNAs (lncRNAs) were directly or indirectly involved in the occurrence and development of tumors, including hepatocellular carcinoma (HCC). Recent studies had found that the expression of lncRNA HAND2-AS1 was downregulated in HCC tissues, but its role in HCC progression is unclear. Ultrasound targeted microbubble destruction mediated gene transfection is a new method to overexpress genes. To study the role of ultrasound microbubbles (UTMBs) mediated HAND2-AS1 in the progression of HCC, in order to provide a new reference for the treatment of HCC. In vitro, we transfected HAND2-AS1 siRNA into HepG2 cells by UTMBs, and detected cell proliferation, apoptosis, invasion and epithelial-mesenchymal transition (EMT) by cell counting kit-8 assay, flow cytometry, Transwell invasion assay and Western blotting, respectively. In addition, we transfected miR-837-5p mimic into UTMBs treated cells and observed the changes of cell behavior. Next, the UTMBs treated HepG2 cells were transfected together with miR-837-5p mimic and tissue inhibitor of matrix metalloproteinase-2 (TIMP2) overexpression vector, and we detected cell proliferation, apoptosis, invasion and EMT. In vivo, we established a mouse model of subcutaneous transplantation of HepG2 cells and observed the effect of HAND2-AS1 silencing on tumor formation ability. We found that UTMBs carrying HAND2-AS1 restricted cell proliferation, invasion, and EMT, encouraged apoptosis, and HAND2-AS1 silencing eliminated the effect of UTMBs. Additionally, miR-873-5p targets the gene HAND2-AS1, which also targets the 3'UTR of TIMP2. And miR-873-5p mimic counteracted the impact of HAND2-AS1. Further, miR-873-5p mimic solely or in combination with pcDNA-TIMP2 had been transformed into HepG2 cells exposed to UTMBs. We discovered that TIMP2 reversed the effect of miR-873-5p mimic caused by the blocked signalling cascade for matrix metalloproteinase (MMP) 2/MMP9. In vivo results showed that HAND2-AS1 silencing significantly inhibited tumor formation in mice. LncRNA HAND2-AS1 promotes TIMP2 expression by targeting miR-873-5p to inhibit HepG2 cell growth and delay HCC progression.

Quantitative proteomics assay reveals G protein-coupled receptor kinase 4-induced HepG2 cell growth inhibition

Background and aimTo investigate the biological effects and putative biological mechanism of G protein-coupled receptor kinase 4 (GRK4) on HepG2 cells. Materials and methodsCell proliferation, cycle, and apoptosis were evaluated by Cell Counting Kit-8 and flow cytometry (FCM) in HepG2 cells infected with either the GRK4-overexpressing lentivirus vector (OE) or the negative control lentivirus vector (NC). The protein profiles and differentially expressed proteins (DEPs) of the OE and NC cells were analyzed and compared using the quantitative proteomics technique, and their function, expression, and probable mechanism were investigated using bioinformatic assays and parallel reaction monitoring (PRM). ResultsHepG2 cells that received the OE grew more slowly than those that received the NC. FCM revealed that, when compared to the NC cells, the OE cells had undergone S-phase cycle arrest, and neither the OE nor NC cells underwent apoptosis. Among the 7006 proteins that were identified by quantitative proteomics, 403 DEPs were examined based on the filtering parameters, with the expressions of 135 being downregulated and 268 being upregulated. In addition to being involved in the peroxisome proliferator-activated receptor (PPAR) signaling pathway, the DEPs were implicated in the biological processes of cell proliferation, cycle, and metabolism. PRM verified the expressions of DEPs that were connected to the PPAR pathway. ConclusionsThis study shows that GRK4 prevents HepG2 cells from proliferating and causes cell cycle arrest in the S-phase, while the PPAR pathway is involved in the regulation of HepG2 cells via GRK4.

Unraveling Interaction of Rhenium-188 Microspheres with Primary Hepatic Cancer Cell: A Breakthrough Study.

Introduction: Hepatocellular carcinoma is a prevalent contributor to global mortality rates. The main palliative treatments are trans-arterial chemoembolization and selective intra-arterial radionuclide therapy. Methods: A novel freeze-dried nonradioactive microsphere kit formulation has been developed, and the behavior and therapeutic potential of 188Re microspheres have been assessed. The microspheres were labeled with fluorescein isothiocyanate (FITC) and 188ReO4-. The uptake of FITC microspheres by HepG2 cells was examined at various time intervals. The impact of 188Re microspheres on cell viability and the mode of cell death were investigated with HepG2 cells using MTT and Annexin FITC-V/propidium iodide (PI) apoptosis assay. Results: The labeling efficiency of microspheres was more than 99% with FITC and 188ReO4-. The maximum uptake of FITC microspheres by HepG2 cells was achieved at 6 h. The exposure to 188Re microspheres has shown a decrease in cellular viability from 77.81% ± 0.015% to 42.03% ± 0.148% at 192 h of incubation (∼11 half-lives). The cellular uptake of 188Re microspheres was 0.255-0.901 MBq. These values were concordant with Annexin FITC-V/PI apoptosis assay. At 192 h, 53.28% ± 0.01% of cells entered the apoptotic phase after treatment with 188Re microspheres, and only 39.34% ± 0.02% of cells remained viable. However, in the cells treated with 188ReO4- alone, 74.86% ± 0.005% of cells were viable, and only 24.75% ± 0.577% of cells were in the early apoptotic phase at 192 h. Conclusion: The data revealed that 188Re microspheres treatment led to significant growth inhibition in HepG2 cells compared with 188ReO4-.

Plumbagin Nanoliposomes Suppress HepG2 Cell Proliferation, Migration, and Invasion by Accelerating miR-16-5p Expression and Inhibiting the VEGFA/EMT Pathway

It has been reported that plumbagin (PL) can inhibit tumor cell growth and induce apoptosis, but the underlying mechanisms remain unclear. In this study, plumbagin nanoliposomes were prepared, and Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2′-deoxyuridine (EdU) assays were performed to determine the effects of PL on HepG2 cell proliferation. Transwell and wound healing assays were also used to investigate the effects of PL on HepG2 cell motility. Additionally, quantitative real-time polymerase chain reaction (qRT–PCR) was carried out to confirm the expression of microRNAs (miRNAs) under PL treatment, in which miR-16-5p showed the most substantially elevated profile among all miRNAs. After transfection with mimics or inhibitors of miR-16-5p or treatment with PL alone, the efficiency of up- and downregulation of miR-16-5p was determined by qRT–PCR in HepG2 cells. Furthermore, EdU, Transwell, and wound healing assays were used to explore the effects of up- and downregulation of miR-16-5p in HepG2 cells. Next, bioinformatic analysis was used to predict the potential target genes of miR-16-5p, along with further validation by dual luciferase reporter assays, qRT–PCR, and western blotting. To investigate the roles of PL and miR-16-5p in vivo, HepG2 cells were infected with lentivirus of miR-16-5p mimics or inhibitor, or the negative control (NC), and stable expression tumor cell lines were established. Then, a tumor mouse model was constructed and PL nanoliposomes were administered to evaluate their therapeutic effect. The volume and weight of tumor were recorded and analyzed and immunohistochemical assays were applied to determine the tumor growth and motility changes. The results showed that PL nanoliposomes suppressed HepG2 cell proliferation and migration in a dose-dependent manner. Mechanistically, PL enhanced miR-16-5p expression and inhibited HepG2 cell growth and migration by targeting the VEGFA/EMT pathway both in vitro and in vivo. Our study demonstrated that PL can inhibit the malignant functions of HepG2 cells by enhancing miR-16-5p expression, which functions as a tumor suppressor gene through targeting the VEGFA/EMT pathway.

Diallyl trisulfide induces pro-apoptotic autophagy via the AMPK/SIRT1 signalling pathway in human hepatocellular carcinoma HepG2 cell line.

Liver cancer is associated with a high mortality rate worldwide. Hepatocellular carcinoma (HCC) constitutes a large proportion of primary liver cancers, and most of its alterations currently remain untreatable. Diallyl trisulfide (DATS), the main chemical constituent of allicin, affects tumour development by regulating cell apoptosis. Allicin-induced autophagy could contribute to apoptosis in HepG2 cells. We rigorously examined the autophagy-related mechanism of allicin-induced apoptosis in HepG2 cells. We treated HepG2 cells with DATS to explore the effect of DATS on pro-apoptotic autophagy in HepG2 cell lines and examine its specific molecular mechanism. HepG2 cells were treated with various concentrations of DATS for 24 and 48 h. Subsequently, cell viability was measured using the cell counting kit-8 (CCK-8) assay and cell clone formation assay. The HepG2 cell apoptosis was measured using Hoechst 33258 staining and western blotting. Autophagy and the AMP-activated protein kinase (AMPK)/NAD-dependent deacetylase sirtuin-1 (SIRT1) signalling pathway were detected using western blotting. Our results indicated that DATS inhibited HepG2 cell growth. Moreover, the ability of DATS to promote apoptosis in HepG2 cells increased with increasing concentration. We verified the phenomenon of DATS-induced autophagy in HepG2 cells and demonstrated that DATS treatment upregulated the protein expression of LC3-II/I. By measuring the expression of potential autophagy stimulators, we documented that DATS could induce pro-apoptotic autophagy by activating the AMPK/SIRT1 signalling pathway. DATS induced pro-apoptotic autophagy via the AMPK/SIRT1 signalling pathway in the human HCC HepG2 cell line. Our findings further implicate allicin as a potential therapeutic agent against liver tumours in clinical settings, providing a basis for combining allicin with an autophagy agonist for treating liver cancer.

Diallyl trisulfide induces pro-apoptotic autophagy via the AMPK/SIRT1 signalling pathway in human hepatocellular carcinoma HepG2 cell line

Background: Liver cancer is associated with a high mortality rate worldwide. Hepatocellular carcinoma (HCC) constitutes a large proportion of primary liver cancers, and most of its alterations currently remain untreatable. Diallyl trisulfide (DATS), the main chemical constituent of allicin, affects tumour development by regulating cell apoptosis. Allicin-induced autophagy could contribute to apoptosis in HepG2 cells. We rigorously examined the autophagy-related mechanism of allicin-induced apoptosis in HepG2 cells. We treated HepG2 cells with DATS to explore the effect of DATS on pro-apoptotic autophagy in HepG2 cell lines and examine its specific molecular mechanism.
 Methods: HepG2 cells were treated with various concentrations of DATS for 24 and 48 h. Subsequently, cell viability was measured using the cell counting kit-8 (CCK-8) assay and cell clone formation assay. The HepG2 cell apoptosis was measured using Hoechst 33258 staining and western blotting. Autophagy and the AMP-activated protein kinase (AMPK)/NAD-dependent deacetylase sirtuin-1 (SIRT1) signalling pathway were detected using western blotting.
 Results: Our results indicated that DATS inhibited HepG2 cell growth. Moreover, the ability of DATS to promote apoptosis in HepG2 cells increased with increasing concentration. We verified the phenomenon of DATS-induced autophagy in HepG2 cells and demonstrated that DATS treatment upregulated the protein expression of LC3-II/I. By measuring the expression of potential autophagy stimulators, we documented that DATS could induce pro-apoptotic autophagy by activating the AMPK/SIRT1 signalling pathway.
 Conclusion: DATS induced pro-apoptotic autophagy via the AMPK/SIRT1 signalling pathway in the human HCC HepG2 cell line. Our findings further implicate allicin as a potential therapeutic agent against liver tumours in clinical settings, providing a basis for combining allicin with an autophagy agonist for treating liver cancer.

A specific anti-cyclin D1 intrabody represses breast cancer cell proliferation by interrupting the cyclin D1-CDK4 interaction.

Cyclin D1 overexpression may contribute to development of various cancers, including breast cancer, and thus may serve as a key cancer diagnostic marker and therapeutic target. In our previous study, we generated a cyclin D1-specific single-chain variable fragment antibody (ADκ) from a human semi-synthetic single-chain variable fragment library. ADκ specifically interacted with recombinant and endogenous cyclin D1 proteins through an unknown molecular basis to inhibit HepG2 cell growth and proliferation. Here, using phage display and in silico protein structure modeling methods combined with cyclin D1 mutational analysis, key residues that bind to ADκ were identified. Notably, residue K112 within the cyclin box was required for cyclin D1-ADκ binding. In order to elucidate the molecular mechanism underlying ADκ anti-tumor effects, a cyclin D1-specific nuclear localization signal-containing intrabody (NLS-ADκ) was constructed. When expressed within cells, NLS-ADκ interacted specifically with cyclin D1 to significantly inhibit cell proliferation, induce G1-phase arrest, and trigger apoptosis of MCF-7 and MDA-MB-231 breast cancer cells. Moreover, the NLS-ADκ-cyclin D1 interaction blocked binding of cyclin D1 to CDK4 and inhibited RB protein phosphorylation, resulting in altered expression of downstream cell proliferation-related target genes. We identified amino acid residues in cyclin D1 that may play key roles in the ADκ-cyclin D1 interaction. A nuclear localization antibody against cyclin D1 (NLS-ADκ) was constructed and successfully expressed in breast cancer cells. NLS-ADκ exerted tumor suppressor effects via blockingthe binding of CDK4 to cyclin D1 and inhibiting phosphorylation of RB. The results presented here demonstrate anti-tumor potential of intrabody-based cyclin D1-targeted breast cancer therapy.

New Schiff base ligand and its novel Cr(III), Mn(II), Co(II), Ni(II), Cu(II), Zn(II) complexes: spectral investigation, biological applications, and semiconducting properties

New Schiff base ligand, derived from antiviral valacyclovir, and its novel Cr(III), Mn(II), Co(II), Ni(II), Cu(II), Zn(II) complexes have been synthesized. By using a variety of analytical and spectroscopic techniques, the type of bonding between the ligand and the metal ions in the recently formed complexes was clarified. The Schiff base ligand act as a bidentate and coordinated with the metal ions through the azomethine-N and the phenolic-O centers, in a mono-deprotonated form. Except for the Zn(II) complex, which displayed a tetrahedral geometry, all complexes displayed octahedral geometry. The TGA findings supported that the stability and decomposition properties of the metal complexes were entirely distinct from one another. The thermogram showed decomposition of all investigated metal complexes above 200 °C in three, four or five steps, and indicated the high thermal stability of these complexes. According to XRD patterns, the particles of these complexes were located at the nanoscale. Moreover, for all the samples analyzed, the TEM images showed uniform and homogeneous surface morphology. The biological activity revealing the high efficiencies of the screened complexes as antibacterial and antitumor agents. The antimicrobial activity of the ligand and its complexes was examined against a variety of pathogenic bacteria and fungi including Escherichia coli, Staphylococcus aureus and Candida albicans. The data obtained revealed that the metal ion in the complexes enhanced the antimicrobial activity compared to the free ligand. The high efficiencies toward S. aureus, E. coli, and C. albicans appeared by Cu(II) complex 23, Ni(II) complex 20, and Ni(II) complex 19, respectively. The antitumor activity of the ligand and its complexes was tested against Hepatocellular carcinoma cell line (HepG-2 cells), the residue 28 which produced after heating the Cu(II) complex 25 at 200 °C for 1 h, exhibited strong inhibition of HepG-2 cell growth. The results of the DNA cleavage investigation demonstrated the ability of investigated Cu(II) complex to degrade DNA. The docking findings showed strong interactions of both the ligand and its examined Cu(II) complex, revealing their ability to cleavage DNA and their potent inhibitory effects on tumor cells. The electrical conductivity study confirmed that the ligand and its investigated complexes had semiconducting properties.

Duguetia pycnastera Sandwith (Annonaceae) Leaf Essential Oil Inhibits HepG2 Cell Growth In Vitro and In Vivo.

Duguetia pycnastera Sandwith (Annonaceae) is a tropical tree that can be found in the Guyanas, Bolivia, Venezuela, and Brazil. In Brazil, it is popularly known as “ata”, “envira”, “envira-preta”, and “envira-surucucu”. In the present work, we investigated the in vitro and in vivo HepG2 cell growth inhibition capacity of D. pycnastera leaf essential oil (EO). The chemical composition of the EO was determined by GC–MS and GC–FID analyses. The alamar blue assay was used to examine the in vitro cytotoxicity of EO in cancer cell lines and non-cancerous cells. In EO-treated HepG2 cells, DNA fragmentation was measured by flow cytometry. The in vivo antitumor activity of the EO was assessed in C.B-17 SCID mice xenografted with HepG2 cells treated with the EO at a dosage of 40 mg/kg. Chemical composition analysis displayed the sesquiterpenes α-gurjunene (26.83%), bicyclogermacrene (24.90%), germacrene D (15.35%), and spathulenol (12.97%) as the main EO constituents. The EO exhibited cytotoxicity, with IC50 values ranging from 3.28 to 39.39 μg/mL in the cancer cell lines SCC4 and CAL27, respectively. The cytotoxic activity of the EO in non-cancerous cells revealed IC50 values of 16.57, 21.28, and >50 μg/mL for MRC-5, PBMC, and BJ cells, respectively. An increase of the fragmented DNA content was observed in EO-treated HepG2 cells. In vivo, EO displayed tumor mass inhibition activity by 47.76%. These findings imply that D. pycnastera leaf EO may have anti-liver cancer properties.

Inhibition of the PINK1-Parkin Pathway Enhances the Lethality of Sorafenib and Regorafenib in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common fatal malignancies and the main cause of cancer-related deaths. The multitarget tyrosine kinase inhibitors (TKIs) sorafenib and regorafenib are systemic therapeutic drugs approved for the treatment of HCC. Here, we found that sorafenib and regorafenib injured mitochondria by inducing mitochondrial Ca2+ (mtCa2+) overload and mitochondrial permeability transition pore (mPTP) opening, resulting in mitochondria-mediated cell death, which was alleviated by cyclosporin A (CsA), an inhibitor of mPTP. Meanwhile, mPTP opening caused PINK1 accumulation on damaged mitochondria, which recruited Parkin to mitochondria to induce mitophagy. Inhibition of autophagy by the lysosomal inhibitor chloroquine (CQ) or inhibition of mitochondrial fission by mdivi-1 aggravated sorafenib- and regorafenib-induced cell death. Moreover, knockdown of PINK1 also promotes sorafenib- and regorafenib-induced cell death. An in vivo study showed that sorafenib and regorafenib inhibited HepG2 cell growth more effectively in PINK1 knockdown cells than in shNTC cells in null mice. Thus, our data demonstrate that PINK1-Parkin-mediated mitophagy alleviates sorafenib and regorafenib antitumor effects in vitro and in vivo.

Mollugin induced oxidative DNA damage via up-regulating ROS that caused cell cycle arrest in hepatoma cells

Mollugin has been proven to have anti-tumor activity. However, its potential anti-tumor mechanism remains to be fully elaborated. Herein, we investigated the growth inhibition of HepG2 cells, as well as the anti-tumor effect of mollugin and its molecular mechanism on H22-tumor bearing mice. In vitro, mollugin was shown to have a strong inhibitory effect on HepG2 cells in a concentration-dependent manner. Mollugin induced S-phase arrest of HepG2 cells, and increased intracellular reactive oxygen species (ROS) levels. Comet assay demonstrated that mollugin induced DNA damage in HepG2 cells, as well as an increase in the expression of p-H2AX. In addition, mollugin induced changes in cyclin A2 and CDK2. However, the addition of antioxidant glutathione (GSH) was able to reverse the effect of mollugin. In vivo, mollugin significantly inhibited tumor growth and reduced the tendency of tumor volume growth in mice. The tumor cell density was found to be decreased in the administration group, and the content of ROS in the tumor tissue significantly increased. The expression of p-H2AX, cyclin A2 and CDK2 were consistent with in vitro results. Mollugin demonstrated anti-hepatocellular carcinoma activity in vitro and in vivo, and its anti-hepatocellular carcinoma activity was found to be related to DNA damage and cell cycle arrest induced by excessive ROS production in cells.

Long noncoding RNA Gas5 induces cell apoptosis and inhibits tumor growth via activating the CHOP-dependent endoplasmic reticulum stress pathway in human hepatoblastoma HepG2 cells.

In recent years, long noncoding RNAs (lncRNAs) have been demonstrated to be important tumor-associated regulatory factors. LncRNA growth arrest-specific transcript 5 (Gas5) acts as an anti-oncogene in most cancers. Whether Gas5 acts as an oncogene or anti-oncogene in hepatocellular carcinoma (HCC) remains unclear. In the present study, the expression and role of Gas5 in HCC were investigated in vitro and in vivo. Lower expression levels of Gas5 were determined in HCC tissues and cells by quantitative reverse transcription-polymerase chain reaction. Overexpressed Gas 5 lentiviral vectors were constructed to analyze their influence on cell viability, migration, invasion, and apoptosis. Fluorescence in situ hybridization was used to identify the subcellular localization of Gas5. Protein complexes that bound to Gas5 were isolated from HepG2 cells through pull-down experiments and analyzed by mass spectrometry. A series of novel Gas5-interacting proteins were identified and bioinformatics analysis was carried out. These included ribosomal proteins, proteins involved in protein folding, sorting, and transportation in the ER, some nucleases and protein enzymes involved in gene transcription, translation, and other proteins with various functions.78 kDa glucose-regulated protein (GRP78) was identified as a direct target of Gas5 by Rip-qPCR and Western blot analysisassay. Gas5 inhibited HepG2 cell growth and induced cell apoptosis via upregulating CHOP to activate the ER stress signaling pathway. Further studies indicated that the knockdown of CHOP by shRNA partially reversed Gas5-mediated apoptosis in HepG2 cells. Magnetic resonance imagingshowed that the ectopic expression of Gas5 inhibited the growth of HCC in nude mice. These findings suggest that Gas5 functions as a tumor suppressor and induces apoptosis through activation of ER stress by targeting the CHOP signal pathway in HCC.

Spectroscopic characterization of Cu(II), Ni(II), Co(II) complexes, and nano copper complex bearing a new S, O, N-donor chelating ligand. 3D modeling studies, antimicrobial, antitumor, and catalytic activities

A new tridentate hydrazone ligand (HL), its Co(II), Ni(II), Cu(II) complexes (in 1:1, 1:2 molar ratios of metal to ligand), and the mixed-ligand Co(II), Ni(II), Cu(II) complexes of the ligand HL, with 8-HQ in 1:1:1 (M:L:8-HQ) stoichiometry, in addition to a nano Cu(II) complex have been synthesized and characterized using physical, analytical and spectral methods. Octahedral geometry was assigned for all investigated complexes except Cu(II) complex 1 which exhibited square planar arrangement. The TGA results suggested the thermal stability of the current complexes. The XRD data indicated that the particles of Cu(II) complex 2 were situated in nano-scale range. Furthemore, the SEM and TEM images of the nano Cu(II) complex 2 showed spherical and sheet-shaped particles with an average size of 48 nm. The geometries of the metal complexes were optimized with respect to the energy taking the 6-311G(p,d) basis set in Gaussian 09 W program in gaseous phase by DFT. The data obtained revealed that all metal complexes are more reactive than the free ligand HL, and the Cu(II) complex 1 has the highest chemical reactivity. The antimicrobial activity study illustrated enhancement in activity of the free ligand upon complex formation., and the Cu(II) complex 1 may be considered as promising antibacterial agent, and as promising antifungal agent. The results of antitumor activity study revealed an inhibition of HepG-2 cell growth for the ligand HL, with enhancement in activity upon complexation, and the Cu(II) complex 1 exhibited the highest cytotoxic activity. The phenoxazinone synthase activity study indicated the ability of both Cu(II) complexes 1 and 2 to catalyze the oxidative dehydrogenation of 2-aminophenol (OAPH) to 2-aminophenoxazine-3-one (APX) under atmospheric conditions and at room temperature.

Structure analysis of ethyl ferulate from Rubus corchorifolius L.f. leaves and its inhibitory effects on HepG2 liver cancer cells

The tender leaves of Rubus corchorifolius L. f. (R. corchorifolius) have been made into herbal tea for relieving coughs in East and Southeast Asia from ancient times. Herein, we obtained an ethyl acetate extract from R. corchorifolius leaves and isolated a major bioactive compound within. Structural analysis of the isolated compound was performed by spectroscopic methods and the compound was identified as ethyl ferulate. Effect of ethyl ferulate on the growth and proliferation of human liver cancer HepG2 cells and normal human liver LO2 cells was determined by MTT assay. We found that ethyl ferulate significantly suppressed the cell proliferation of HepG2 cells (IC50 26.4 μM), while it did not cause cytotoxicity on LO2 cells. In addition, flow cytometry analysis demonstrated that ethyl ferulate inhibited HepG2 cell growth via inducing cellular apoptosis. Specifically, ethyl ferulate reduced the electrical potential of the mitochondrial membrane and increased the concentration of calcium ions inside cells. Moreover, these changes were accompanied by increases in the expression of the pro-apoptotic proteins, such as cleaved caspase-3 and Bax. In contrast, cell morphology and growth of LO2 cells remained unaffected. In conclusion, for the first time, our observations demonstrated that ethyl ferulate possesses great potential as an effective and safe antitumor agent for cancer chemoprevention in humans.

Silver Nanoparticles Stabilized with Phosphorus-Containing Heterocyclic Surfactants: Synthesis, Physico-Chemical Properties, and Biological Activity Determination.

Phosphorus-containing heterocyclic cationic surfactants alkyldimethylphenylphospholium bromides with the alkyl chain length 14 to 18 carbon atoms were used for the stabilization of silver nanodispersions. Zeta potential of silver nanodispersions ranges from +35 to +70 mV, which indicates the formation of stable silver nanoparticles (AgNPs). Long-chain heptadecyl and octadecyl homologs of the surfactants series provided the most intensive stabilizing effect to AgNPs, resulting in high positive zeta potential values and smaller diameter of AgNPs in the range 50–60 nm. A comparison with non-heterocyclic alkyltrimethylphosphonium surfactants of the same alkyl chain length showed better stability and more positive zeta potential values for silver nanodispersions stabilized with heterocyclic phospholium surfactants. Investigations of biological activity of phospholium-capped AgNPs are represented by the studies of antimicrobial activity and cytotoxicity. While cytotoxicity results revealed an increased level of HepG2 cell growth inhibition as compared with the cytotoxicity level of silver-free surfactant solutions, no enhanced antimicrobial action of phospholium-capped AgNPs against microbial pathogens was observed. The comparison of cytotoxicity of AgNPs stabilized with various non-heterocyclic ammonium and phosphonium surfactants shows that AgNPs capped with heterocyclic alkyldimethylphenylphospholium and non-heterocyclic triphenyl-substituted phosphonium surfactants have the highest cytotoxicity among silver nanodispersions stabilized by the series of ammonium and phosphonium surfactants.

Identification of Ethyl Ferulate From Rubus corchorifolius L.F. Leaves and Its Inhibitory Effects on HepG2 Liver Cancer Cells

Abstract Objectives Rubus corchorifolius L.f., also known as raspberry, March bubble, milk bubble, etc., belongs to the genus Rubus L. and the family Rosaceae. The tender leaves of Rubus corchorifolius L. f. have been made into herbal tea for drinking and relieving coughs in China since ancient times. In this study, we isolated a bioactive compound from the ethyl acetate extract of Rubus corchorifolius L.f. leaves and investigated its protective effects in HepG2 human liver cancer cell line and the underlying molecular mechanisms. Methods Structural analysis of the isolated compound was performed by spectroscopic methods and the compound was identified as ethyl ferulate. Efficacy and mechanisms of ethyl ferulate on the proliferation of HepG2 human liver cancer cells and LO2 normal human liver cells was determined by cell viability, cell morphology, apoptosis, mitochondrial membrane potential, Ca2 + release, and immunoblotting analyses. Results We found that ethyl ferulate significantly suppressed the cell proliferation of HepG2 cells (IC50 26.4 μM), while it did not cause cytotoxicity on LO2 cells. In addition, flow cytometry analysis demonstrated that ethyl ferulate inhibited HepG2 cell growth via inducing cellular apoptosis. Specifically, ethyl ferulate reduced the electrical potential of the mitochondrial membrane and increased the concentration of calcium ions inside cells. Moreover, these changes were accompanied by increases in the expression of the pro-apoptotic proteins, such as cleaved caspase-3 and Bax. In contrast, cell morphology and growth of LO2 cells remained unaffected. Conclusions In conclusion, our findings demonstrated, for the first time, that ethyl ferulate possesses great potential as an effective and safe antitumor agent for cancer chemoprevention in humans. Funding Sources This study was supported by the University of International Cooperation in Science and Technology Innovation Platform Project of Guangdong (2013gjhz0003). XC was partly supported by a scholarship from South China Agriculture University.

Novel nanoparticle-size metal complexes derived from acyclovir. Spectroscopic characterization, thermal analysis, antitumor screening, and DNA cleavage, as well as 3D modeling, docking, and electrical conductivity studies

Novel nano-sized Cr(III), Mn(II), Co(II), Ni(II), Cu(II) and Zn(II) complexes derived from acyclovir (ACV) were synthesized in different media. The synthesized metal complexes were characterized by different analytical and spectroscopic techniques. The ACV behaves as a bidentate (NO) chelating ligand. All complexes showed octahedral geometry, except Zn(II) complex which exhibited a tetrahedral geometry. The TGA results confirmed thermally stability of these complexes. The XRD patterns and TEM images indicated that the particles of the investigated complexes were situated in nano-scale range. The data obtained from the computational estimation agree well with the experimental results. The results of antimicrobial screening revealing enhancement in activity upon complex formation. All investigated compounds exhibited an inhibition of HEPG-2 cell growth. Additionally, the antitumor activity of the nano-scale complexes were enhanced after heating of them at 200 °C for 1 hour. The DNA cleavage study showed that the Cu(II) complex XXVII may be able to induce cellular death in cancer cells through apoptosis/necrosis pathways. The docking analysis revealed that the nano-sized Cu(II) complex V has stronger binding affinity to DNA than the free ligand ACV. Furthermore, the electrical conductivity measurements indicated that the ACV and its investigated nano-sized complexes behave as semiconductors, and the hopping model is applicable to describe the mechanism of conduction process.

Comment on: an orally antitumor chalcone hybrid inhibited HepG2 cells growth and migration as the tubulin binding agent.

Recently we read a paper in Investigational New Drugs "An orally antitumor chalcone hybrid inhibited HepG2 cells growth and migration as the tubulin binding agent". Chalcone hybrid 9, a novel chalcone derivative, may be a promising agent for the treatment of hepatocellular carcinoma. However, there are some problems in this paper that are worthy of comment. Human hepatocellular carcinoma HepG2 cells from Shanghai Research Science Limited Company could generate xenograft in nude mice and chalcone hybrid 9 suppressed the growth of HepG2 tumor. However, according to the description of cell bank of Chinese Academy of Science and ATCC, HepG2 cells are no tumorigenic. Similarly, our lab also confirmed that HepG2 cells cannot demonstrate tumorigenic ability in nude mice. Therefore, this discrepancy raised our concern about HepG2 xenograft in the paper.

α-lipoic acid (α-lip) modification on surface of nano-scaled zeolitic imidazole Framework-8 for enhanced drug delivery

Curcumin (CCM) is a natural ingredient with anti-cancer activity. However, the application of CCM is limited in clinical medicine due to its high hydrophobicity and low bioavailability. Hence, it is necessary for development of a feasible strategy for efficient drug delivery and cancer cell-selective targeting. In recent years, nano-scaled metal-organic frameworks (NanoMOFs) have received much attention as an attractive drug delivery carrier due to their unique features of their various sizes, adjustable pore structures, high stability, and easy surface modification. Here, we report a novel composite nanomaterial (denoted as ZIF-8&α-lip) for CCM loading by one-pot method. This synthetic method is simple and rapid, and the final product (CCM@ZIF-8&α-lip) can be obtained within 20 ​min. In addition, the obtained CCM@ZIF-8&α-lip shows greater growth inhibition of HepG2 cells in comparison with CCM@ZIF-8. Therefore, ZIF-8&α-lip can be used as an ideal drug carrier and propose a new strategy to improve nanoMOFs performance for efficient drug delivery.

Water-Soluble Pristine C60 Fullerene Inhibits Liver Alterations Associated with Hepatocellular Carcinoma in Rat

Excessive production of reactive oxygen species is the main cause of hepatocellular carcinoma (HCC) initiation and progression. Water-soluble pristine C60 fullerene is a powerful and non-toxic antioxidant, therefore, its effect under rat HCC model and its possible mechanisms were aimed to be discovered. Studies on HepG2 cells (human HCC) demonstrated C60 fullerene ability to inhibit cell growth (IC50 = 108.2 μmol), to induce apoptosis, to downregulate glucose-6-phosphate dehydrogenase, to upregulate vimentin and p53 expression and to alter HepG2 redox state. If applied to animals experienced HCC in dose of 0.25 mg/kg per day starting at liver cirrhosis stage, C60 fullerene improved post-treatment survival similar to reference 5-fluorouracil (31 and 30 compared to 17 weeks) and inhibited metastasis unlike the latter. Furthermore, C60 fullerene substantially attenuated liver injury and fibrosis, decreased liver enzymes, and normalized bilirubin and redox markers (elevated by 1.7–7.7 times under HCC). Thus, C60 fullerene ability to inhibit HepG2 cell growth and HCC development and metastasis and to improve animal survival was concluded. C60 fullerene cytostatic action might be realized through apoptosis induction and glucose-6-phosphate dehydrogenase downregulation in addition to its antioxidant activity.