Liver Cancer Cell Line HepG2 Research Articles

Anthocyanin-loaded polymeric dextran nanoparticle as an anti-proliferative agent on human liver carcinoma cells

In this communication, anthocyanin-loaded dextran nanoparticles from Clitoria ternatea was synthesized and characterized to test its anti-proliferative activity on the human HepG2 liver cancer cell lines. By using dextran as an encapsulant polymer, the nanoparticles appeared to be spherical, with an average size of 45.5 ± 11 nm. The surface charge of the anthocyanin-loaded dextran nanoparticle was -4.39 mV, which slightly relative to free anthocyanin (-4.46 mV), which indicate good dispersion stabilities. The Fourier transform infrared analysis showed that the anthocyanins from C. ternatea was successfully encapsulated in dextran nanoparticles. Overall, the percentage of drug encapsulation efficiency was 3.03%. Based on the stability test, the anthocyanin-loaded dextran nanoparticle showed significantly better color stability index compared to free anthocyanin, particularly at the presence of light and temperature of 37°C and 50°C. In the anti-proliferation assay on HepG2 liver cancer cell lines, the viability of the cancer cells was significantly reduced after treatment with the anthocyanin-loaded dextran nanoparticle. The anti-proliferation activities of the nanoparticles were significantly better than free anthocyanin. Our findings revealed the ability of the anthocyanin-loaded dextran nanoparticle, in particular from C. ternatea, as an effective anti-proliferative agent against cancer cells. Nanoencapsulation with dextran significantly improve the efficacy and stability of the anthocyanins. Further investigations should be done to evaluate the in vivo efficacy.

Cytotoxic activity, Molecular docking study and Phytochemical investigation on Cichorium intybus herb

The purpose of this study is to evaluate the cytotoxic activity of Cichorium intybus herb as well as to identify the molecular mechanism of the cytotoxicity. In addition, it aims to investigate its phytoconstituents that are responsible for the bioavailability. The cytotoxic activity of Cichorium intybus herb was assayed by SRB (Sulforhodamine B) assay against ovarian cancer cell line (SKOV-3), liver cancer cell line (HepG2) and prostate cancer cell line (PC-3). The effect on tubulin polymerization was studied to identify the mechanism of cytotoxicity. The binding affinity to the target molecule was examined by docking study. In addition, two flavonoidal compounds were isolated and identified by different spectroscopic methods. The results showed that the methanol extract of Cichorium intybus herb as well as the isolated compounds (myricetin and pinobanksin) possessed a potent cytotoxicity against HepG2 (IC50 =0.95, 4.26 and 7.23 μg/mL), respectively, moderate cytotoxicity against PC-3 (IC50 =25.34, 36.24 and 42.53 μg/mL), respectively, and weak cytotoxicity against SKOV-3 (IC50 >100 μg/mL) for all tested samples. Molecular docking analysis confirmed that both of the isolated compounds showed high binding affinity to colchicine binding site of tubulin microtubules, supported the high cytotoxicity of these compounds.

Quinoline-pyrimidine hybrid compounds from 3-acetyl-4-hydroxy-1-methylquinolin-2(1H)-one: Study on synthesis, cytotoxicity, ADMET and molecular docking

Some novel 2-amino-6-aryl-4-(4′-hydroxy-N-methylquinolin-2′-on-3′-yl)pyrimidines have been synthesized from α,β-unsaturated ketones of 3-acetyl-4-hydroxy-N-methylquinolin-2-one by reaction of corresponding α,β-unsaturated ketones with guanidine hydrochloride. The purity and structure of the obtained products have been confirmed by thin layer chromatography, IR, 1H NMR, 13C NMR, HSQC, HMBC and MS spectra. All the synthesized of 3-(2-amino-6-arylpyrimidin-4-yl)-4-hydroxy-1-methylquinolin-2(1H)-ones 6a-i were screened for their in vitro cytotoxic activity against human hepatocellular carcinoma HepG2 and squamous cell carcinoma KB cancer lines. Compounds 6b and 6e had the best activity in the series, with IC50 values equal to 1.33 μM. Compounds 6a-g exhibited weak or insignificant activity with liver cancer cell lines HepG2, while compounds 6a and 6g had more powerful activity in this sequence, with IC50 values equal to 47.99 and 89.38 μM, respectively. ADMET properties showed that compounds 6b, 6e, and 6f possessed the drug-likeness behavior. Cross-docking results indicated that two hydrogen bonding interactions in the binding pocket, as potential ligand binding hot-spot residues for compounds 6b and 6e, may be one of the mechanisms of action responsible for the higher cytotoxic effect on HepG2 and KB cells.

Biocompatible Delivery System for Metformin: Characterization, Radiolabeling and In Vitro Studies.

In recent years, the uses of nanotechnology in medicine have an increasing potential as an effective nanocarrier system. These systems are improved with the purpose of maximizing therapeutic activity and minimizing undesirable side-effects. Moreover, radiolabeled nanoparticles can be used as agents for diagnosis and therapeutic purposes in clinical applications. They have three main components: the core, the targeting biomolecule, and the radionuclide. It is aimed to synthesize Metformin (MET) loaded Solid Lipid Nanoparticles (MET-SLN) and radiolabeled with technetium-99m tricarbonyl core. The structure of synthesized nanoparticles was characterized by Fourier Transform Infrared Spectroscopy (FTIR). The particle size and morphology of nanoparticles were examined by Dynamic Light Scattering (DLS), and Scanning Electron Microscope (SEM). Quality control studies of radiolabeled MET-SLN [99mTc(CO)3-MET-SLN] were performed by High-Performance Liquid Radiochromatography (HPLRC) and Thin Layer Radiochromatography (TLRC). The radiolabeling yield of [99mTc(CO)3-MET-SLN] was found to be 88%. In vitro studies have been performed on cancer lines(MCF7, MDA-MD-231 breast, and HEPG2 liver cancer cells) to determine the biological behavior of 99mTc(CO)3-MET-SLNs. The results showed that higher uptake values were observed on estrogen-positive MCF7 breast cancer cell line according to estrogen negative MDA-MB-231 breast cancer and HEPG2 liver cancer cell lines.

Cytotoxic Effects of Moringa oleifera Leaf Extract on Human Hepatoma Cell Line HepG-2

Background: Over the past decades, little attention is paid to using herbal remedies to improve various diseases, especially cancer. Moringa oleifera (MO) is one of the herbs with numerous medicinal properties. Objectives: The current study aimed to evaluate the cytotoxicity effects of methanolic leaf extract on human liver cancer HepG2 cell lines. Methods: First methanolic extract of Moringa oleifera flavonoid compounds were measured, and then quercetin was identified as one of the flavonoid compounds in methanolic leaf extract by high-performance liquid chromatography (HPLC). HepG-2 cell line were treated with (0, 30, 40, 50, 60, and 70 µg/mL) methanolic extract of MO, quercetin (50 µg/mL), and doxorubicin (1 µg/mL). After 48 hours, the cytotoxic activity of various concentrations of MO methanolic extract against the liver cancer cell line was assessed using an MTT assay. Results: In this study, the total flavonoids and quercetin in the leaves of MO was, respectively, 3.69 (mg/g) and 0.064 (mg/g) of dry weight. Methanolic leaf extract demonstrated a significant effect (IC50 = 12.89 µg/mL) on HepG2 cell lines. Conclusions: Evaluation of cytotoxicity effects showed that HepG2 viability methanolic extract was concentration-dependent and decreased with increasing concentration of the extract. Comparison of the effect of the extract with doxorubicin and pure quercetin showed that the effect of the concentration of 70 µg/mL methanolic extract and pure quercetin and doxorubicin on liver cancer cells were similar. Also, the results showed that methanolic leaf extract has cytotoxic effects on the HepG-2 cell line and inhibits the growth of liver cancer cells.

DNA/bovine serum albumin binding and cytotoxicity of transition metal ternary complexes based on sulfamethazine and bromazepam drugs

Similar to the antibiotic mixing strategies, which depend on reducing the worth of diffusion of the resistant bacteria to new host, in this contribution, we explicit new mixed‐ligand complexes of the type [M(SMZ)2(BZM)] (M = Co(II), Ni(II) and Cu(II)) based on two drugs: sulfamethazine (SMZ) and bromazepam (BZM). The novelty aspect of the present work relies on combining two drugs in a single molecule via the synthesis of the ternary complexes. The ternary complexes were characterized by spectroscopic and analytical tools with the aid of the quantum chemical calculations. Copper(II) complex has the highest DNA intrinsic binding constant (2.20 × 105 M−1), induces extensively DNA cleavage compared to the other complexes, and exhibits the highest cytotoxicity (IC50 = 0.67 μM) against human liver cancer cell line (HepG2). Exciting, Cu(II) compound exhibits no toxicity to the nonmalignant human foetal lung fibroblast (WI‐38). Alternatively, the low cytotoxicity of the Co(II) complex correlates well with the high affinity of the compound to human serum albumin (Kb = 4.58 × 105) compared to 1.93 × 105 and 2.14 × 105 M−1 for Ni (II) and Cu (II) complexes, respectively.

FAM172A affects cell proliferation and apoptosis not by targeting β-tubulin in HepG2 cells.

BackgroundMicrotubules pull chromosomes apart during cell mitosis and take part in cell division, Inhibiting the formation of spindle microtubules during mitosis has become one of the current anti-tumor research strategies. Earlier studies have found that the family with sequence similarity 172, member A (FAM172A) can significantly inhibit the proliferation of human colorectal cancer cell line LOVO cells and promote apoptosis. The purpose of this study was to investigate the biological effects of FAM172A on liver cancer cells and the interaction mechanism with tubulin.MethodsUse STRING software predicted the interactions between FAM172A and β-tubulin, and verify by immunoprecipitation. Real-Time qPCR was used to determine the expression levels of β-tubulin in liver cancer cell line HepG2, western blot was performed to detect protein expression levels. Immunofluorescence experiment to detect the distribution, shape and the dynamic behavior of depolymerization-aggregation of β-tubulin in cells. MTT, wound healing and Transwell assay were employed to determine cell proliferation, migration and invasion respectively. Flow cytometry was conducted to determine cell cycle and apoptosis.ResultsThere is no interactions between FAM172A and β-tubulin. We determined that when FAM172A was up-regulated or down-regulated, the mRNA and protein levels of β-tubulin did not change significantly (P>0.05). Furthermore, the distribution, shape of β-tubulin in cells, and the dynamic behavior of depolymerization-aggregation was not affected. After FAM172A overexpression, the migration and invasion of HepG2 cells were significantly inhibited (P<0.05), the cell proliferation was also significantly inhibited (P<0.05) and was time-dependent. The HepG2 cells had apparent S phase arrest and apoptosis (P<0.05). After interfering with FAM172A, the opposite result will appear.ConclusionsThe results show that FAM172A may be a new tumor suppressor gene, which has a specific role in cell cycle control and cell proliferation, but the specific mechanism of action has not been explained in this study and needs further exploration.

Gold Nanoparticles from Ostruthin: Green Synthesis, a Stable Mechanism, Antitumor, and Antibacterial Activity

The Ostruthin was extracted and identified its structure from rhizomes of Luvunga scandens located at Ta Cu Mountain, Binh Thuan province, Vietnam for the first time. Gold nanoparticles (AuNPs) were synthesized by the green method. The AuNPs acted as antitumor against breast cancer cell line (MCF-7), human liver cancer cell line (HepG2), and Non-Small Cell Lung (NCI-H460). They showed the potential antitumor activity against MCF-7 with the IC50 value of 65.47�3.09 �M. The antitumor activity of the AuNPs was also compared with the extracted constituents from the root of Luvunga Scandens in a previous article. The AuNPs were exposed to high antitumor activity against MCF-7 and Hep G2, human cancer cell lines. The AuNPs have also been tested the antibacterial activity and shown the moderate antibacterial activity on both Salmonella enterica and Bacillus subtilis at a concentration of 0.25 mM.

Water-soluble galactosamine derivative of β-cyclodextrin as protective ligand and targeted carrier for delivery of toxic anthracycline drug

β-cyclodextrin modified with an electron-rich aromatic triazole linker and targeting moiety (galactosamine) was synthesized and studied as a carrier for the anticancer drug, doxorubicin (DOX), with the aim of targeting the pathological cells, reducing the cardiotoxic side effects and increasing the binding of the drug to DNA. The β-cyclodextrins modified with galactosamine (βCDGAL) are non-toxic and highly soluble in aqueous medium compared to the native βCD and βCD modified only with aromatic moiety, such as triazole linker. Molecular modelling and NMR study gave a deeper insight into the ligand structure, providing an explanation for its increased solubility, and the drug-ligand interactions. The triazole linker strengthened the drug binding and introduced pH dependence of the complex stability constants for βCDGAL derivative, as confirmed by the voltammetry measurements. Spectroscopic studies have shown that entrapment of the DOX in βCDGAL cavity reduces the stability constant of the DOX:Fe(III) complex responsible for the production of cardiotoxic reactive oxygen species and additionally supports the binding of the drug to the double strand DNA. The MTT assay and confocal microscopy results showed that despite encapsulation of the drug in the cyclodextrin molecule, its cytotoxic effect on the liver cancer cell line (HepG2) is comparable to that of the free, non-protected drug.

Jaceidin Flavonoid Isolated from Chiliadenus montanus Attenuates Tumor Progression in Mice via VEGF Inhibition: In Vivo and In Silico Studies.

Phytochemical study of Chiliadenus montanus aerial parts afforded six compounds; Intermedeol (1), 5α-hydroperoxy-β-eudesmol (2), 5,7-dihydroxy-3,3’,4’-trimethoxyflavone (3), 5,7,4’-trihydroxy-3,6,3’-trimethoxyflavone (jaceidin) (4), eudesm-11,13-ene-1β,4β,7α-triol (5) and 1β,4β,7β,11-tetrahydroxyeudesmane (6). These compounds were identified based on their NMR spectral data. The isolated compounds were tested for their cytotoxicity against liver cancer cell line (HepG2) and breast cancer cell line (MCF-7). Jaceidin flavonoid (4) exhibited the highest cytotoxic effect in vitro. Therefore, both of jaceidin and C. montanus extract were evaluated for their in vivo anti-tumor activity against Ehrlich’s ascites carcinoma (EAC). Compared to control group, jaceidin and C. montanus extract decreased the tumor weight, improved the histological picture of tumor cells, lowered the levels of VEGF and ameliorate the oxidative stress. Molecular docking and in silico studies suggested that jaceidin was a selective inhibitor of VEGF-mediated angiogenesis with excellent membrane permeability and oral bioavailability.

In Vitro Anticancer Evaluation of Some Synthesized 2H-Quinolinone and Halogenated 2H-Quinolinone Derivatives as Therapeutic Agents.

Searching for new cytotoxic agents with apoptosis induction may represent a viable strategy for cancer treatment to overcome the increased resistance to available anticancer agents. The purpose of the current study was aimed at preparation and anticancer evaluation of two new series of 2H-quinolinone and halogenated 2H-quinolinone derivatives against two cancer cell lines. Two new series of 2H-quinolinone and halogenated 2H-quinolinone derivatives were prepared and screened for their cytotoxicity against breast MCF-7 and liver HepG-2 cancer cell lines as well as normal breast MCF-10a. The tested molecules revealed good cytotoxicity and selectivity toward cancer cell lines relative to normal cells. These compounds were analyzed by DNA flow cytometry on MCF-7 cells. They were found to cause G2/M phase arrest and induced apoptosis at the pre-G1 phase. In addition, increased caspase 3/7 activity and decreased osteopontin expression verified the apoptotic activity. The potent compounds discovered in this study can be a hit for the discovery of new cytotoxic agents and are worthy of further investigation.

Trimetazidine improves hepatic lipogenesis and steatosis in non‑alcoholic fatty liver disease via AMPK‑ChREBP pathway.

Clinical studies have demonstrated that trimetazidine (TMZ) possesses a synergistic hypolipidemic effect together with statins, but the underlying mechanism remains to be elucidated. The present study aimed to investigate the role of TMZ in non-alcoholic fatty liver disease (NAFLD). By investigating the TMZ treatment of NAFLD, it was identified that high-fat diet (HFD) mice exhibit significant changes in several physiologic indices, including body weight, plasma lipids and glucose tolerance. Notably, hepatocyte bullous steatosis and fibrosis in HFD mice are greatly attenuated by 8 weeks of TMZ treatments. The results of the present study also indicated that the expression of carbohydrate-responsive element-binding protein (ChREBP), fatty acid synthase and acetyl-CoA carboxylase were all significantly reduced in the HFD + TMZ group compared with the HFD group. In order to confirm the hypothesis in vitro, the palmitate-treated liver cancer cell line (HepG2) was employed and similar results were obtained in TMZ-treated HepG2 cells. Furthermore, TMZ markedly upregulated the AMP-activated protein kinase (AMPK) signaling pathway and reduced the expression of forkhead box O1 (FOXO1) in the cells, while these effects controlled by TMZ were abolished by the AMPK inhibitor Compound C. The present study reported that knockdown of FOXO1 expression by FOXO1 small interfering RNA resulted in a reduction of ChREBP protein expression and post-transcriptional activity. In summary, for the first time, to the best of the authors' knowledge, the present study revealed a novel role of TMZ in hepatic steatosis; TMZ ameliorated ChREBP-induced de novo lipogenesis by activating the AMPK-FOXO1 pathway.

A novel Romani microdeletion variant in the promoter sequence of ASS1 causes citrullinemia type I

BackgroundCitrullinemia type I (CTLN1, MIM #215700) is an autosomal recessive urea cycle disorder caused by deficiency of argininosuccinate synthase (ASS). CTLN1 is characterized by life-threatening hyperammonemia and risk for resulting neurocognitive impairments. The diagnosis of CTLN1 is confirmed by the identification of biallelic pathogenic variants in the ASS1 gene. However, there are a small percentage of CTLN1 patients with a characteristic biochemical phenotype without identifiable variants in ASS1. We describe the molecular characterization of two related Romani children with biochemically diagnosed CTLN1, whose clinical genetic testing failed to detect any pathogenic variant in ASS1. MethodsGenomic DNA was extracted from peripheral blood lymphocytes collected from both patients. Sanger sequencing was performed after PCR amplifications of 5′- and 3′-untranslated regions of the ASS1 gene. A luciferase reporter assay was performed using the human malignant melanoma A2058 cell line and the human liver cancer cell line HepG2. ResultsWe interrogated the non-coding regions of ASS1 by targeted PCR amplification and identified a homozygous 477-bp microdeletion in the promoter region of the ASS1 gene in both patients. Heterozygosity of the variant was confirmed in their parents. Sanger sequencing confirmed the microdeletion contained the entire sequence of the non-coding exon 1 of ASS1 that includes promoter elements of GC-box, E-box, AP2-binding site, and TATA-box. Luciferase reporter assay using an expression plasmid containing the wild-type or mutant ASS1 sequences showed robust reporter expression from the wild-type sequence and significantly reduced expression driven by the mutant insert (3.6% in A2058 cells and 3.3% in HepG2 cells). These findings were consistent with the hypothesis that the microdeletion identified in the patients disrupted an essential promoter element and resulted in deficiency of ASS1 mRNA expression. ConclusionsThis is the first report of CTLN1 patients caused by a Romani microdeletion variant affecting the non-coding upstream sequence of ASS1. Ablation of the promoter sequence can cause CTLN1 by the reduction of ASS1 expression. Currently available clinical sequencing methods usually do not cover the promoter sequence including the non-coding exon of ASS1, highlighting the importance of evaluating this region in genetic testing for CTLN1.

Apatinib Inhibits the Invasion and Metastasis of Liver Cancer Cells by Downregulating MMP-Related Proteins via Regulation of the NF-κB Signaling Pathway.

Objective We aimed to investigate whether apatinib has an inhibitory effect on the invasion and metastasis of liver cancer in vitro. Methods The anti-invasion and antimetastasis effects of apatinib in HepG2, Hep3B,Huh7 and SMMC-7721 liver cancer cell lines were tested by the wound-healing and transwell invasion assays. Real-time PCR and Western blot were used to detect the influence of apatinib on the gene expression of MMPs, TIMPs, and constituents of the NF-κB signaling pathway in Hep3B and HepG2 liver cell lines. Results Apatinib has a significant inhibitory effect on the metastasis and invasion of liver cancer cells. The expression levels of MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, MMP-10, MMP-11, and MMP-16 were downregulated, while the expression levels of TIMP-3 and TIMP-4 were upregulated by apatinib treatment at both the mRNA and protein levels. The phosphorylation of IκBα and NF-κB p65 was significantly reduced compared with that in the control group. Conclusions Apatinib inhibits the invasion and metastasis of human liver cancer cells by downregulating the expression of MMP-related genes. This may be achieved by inhibiting the activation of the NF-κB signaling pathway.

Cytotoxic effects of individual and binary combinations of zearalenone and ochratoxin a on liver

Abstract Co-contamination with mycotoxin represents a serious concern for human and animal health. In this in vitro study, we investigated the combined effects of ZEA and OTA, mycotoxins which frequently contaminate cereals, in binary mixtures on the viability of human liver cancer cell line (HepG2). Cell viability was assessed after 24 h using a neutral red assay. An antagonistic effect was observed for binary toxins combinations affecting 25% of cell viability (CI=4.18), which turn into a synergistic effect as followed: slight at IL50 (CI=1.51), moderate at IL75 (CI=0.554) and strong at IL90 (CI=0.203). In conclusion, our results show an important additive and even synergistic cytotoxic effect of two commonly occurred mycotoxins: zearalenone and ochratoxin when they are present simultaneously in food or feed. The co-exposure to mycotoxins lead to a higher toxicity than the exposure to single toxin. Our study provides important data for mycotoxins risk assessment. In this context, a re-evaluation of the guidance levels for mycotoxins will be required in the future, in order to reduce the health risk associated with the possible consumption of mycotoxin co-contaminated food or feed.

Bioprospecting a native silver-resistant Bacillus safensis strain for green synthesis and subsequent antibacterial and anticancer activities of silver nanoparticles

Green nanomaterials have gained much attention due to their potential use as therapeutic agents. The present study investigated the production of silver nanoparticles (AgNPs) from a silver-resistant Bacillus safensis TEN12 strain, which was isolated from metal contaminated soil and taxonomically identified through 16S rRNA gene sequencing. The formation of AgNPs in bacterial culture was confirmed by using UV–vis spectroscopy with an absorption peak at 426.18 nm. Fourier transform infrared (FTIR) spectroscopy confirmed the involvement of capping proteins and alcohols for stabilization of AgNPs. Moreover, X-ray diffraction analysis (XRD), scanning and transmission electron microscopy (SEM and TEM) confirmed the crystalline nature and spherical shape of AgNPs with particle size ranging from 22.77 to 45.98 nm. The energy dispersive X-ray spectroscopy (EDX) revealed that 93.54% silver content is present in the nano-powder. AgNPs showed maximum antibacterial activity (20.35 mm and 19.69 mm inhibition zones) at 20 µg mL−1 concentration against Staphylococcus aureus and Escherichia coli, respectively and significantly reduced the pathogen density in broth culture. Furthermore, AgNPs demonstrated significant anticancer effects in the human liver cancer cell line (HepG2) in MTT assay, whereas, no cytotoxic effects were demonstrated by AgNPs on normal cell line (HEK293). The present study suggests that the biogenic AgNPs may substitute chemically synthesized drugs with wider applications as antibacterial and anticancer agents.

In vitro cytotoxic activity of thiazole-indenoquinoxaline hybrids as apoptotic agents, design, synthesis, physicochemical and pharmacokinetic studies.

In this study, anti-proliferative effects of twenty-seven indeno[1,2-b]quinoxalin-11-one derivatives were investigated in three human cancer cell lines, namely: the colon cancer cell line HCT-116, the liver cancer cell line HepG-2, and the breast cancer cell line MCF-7. Among them, 5, 6, 13, 14a, b and 15d-f derivatives displayed excellent anti-proliferative activities against the three tested cell lines compared to the reference standard Imatinib. Therefore, they were selected for further studies. First, to ensure the safety of our hits, investigation of the IC50 values on normal human cells (WI-38) was executed indicating that, they are highly selective (IC50>107μM) in their cytotoxic effect. Second, the induction of apoptosis by these active compounds was achieved by down-regulation of Bcl-2 and up-regulation of BAX and caspase-3. Further investigations have shown that 14b and 15f, the most potent derivatives, induced cell cycle arrest at G2/M phase. Moreover, in silico evaluation of ADME properties indicated that all the potent compounds are orally bioavailable with no permeation to the blood brain barrier.

Phytogenic Synthesis of Nickel Oxide Nanoparticles (NiO) Using Fresh Leaves Extract of Rhamnus triquetra (Wall.) and Investigation of Its Multiple In Vitro Biological Potentials.

Chemically nickel oxide nanoparticles (NiONPs) involve the synthesis of toxic products, which restrict their biological applications. Hence, we developed a simple, eco-friendly, and cost-efficient green chemistry method for the fabrication of NiONPs using fresh leaf broth of Rhamnus triquetra (RT). The RT leaves broth was used as a strong reducing, capping, and stabilizing agent in the formation of RT-NiONPs. The color change in solution from brown to greenish black suggests the fabrication of RT-NiONPs which was further confirmed by absorption band at 333 nm. The synthesis and different physicochemical properties of RT-NiONPs were investigated using different analytical techniques such as UV-Vis (ultraviolet−visible spectroscopy), XRD (X-ray powder diffraction), FT-IR (Fourier-transform infrared spectroscopy), SEM (scanning electron microscopy), TEM (transmission electron microscopy), EDS (energy-dispersive X-ray spectroscopy), DLS (dynamic light scattering) and Raman. Further, RT-NiONPs were subjected to different in vitro biological activities and revealed distinctive biosafe and biocompatibility potentials using erythrocytes and macrophages. RT-NiONPs exhibited potential anticancer activity against liver cancer cell lines HUH7 (IC50: 11.3 µg/mL) and HepG2 (IC50: 20.73 µg/mL). Cytotoxicity potential was confirmed using Leishmanial parasites promastigotes (IC50: 27.32 µg/mL) and amastigotes (IC50: 37.4 µg/mL). RT-NiONPs are capable of rendering significant antimicrobial efficacy using various bacterial and fungal strains. NiONPs determined potent radical scavenging and moderate enzyme inhibition potencies. Overall, this study suggested that RT-NiONPs can be an attractive and eco-friendly candidate. In conclusion, current study showed potential in vitro biological activities and further necessitate different in vivo studies in various animal models to develop leads for new drugs to treat several chronic diseases.

Kaempferol-Mediated Sensitization Enhances Chemotherapeutic Efficacy of Sorafenib Against Hepatocellular Carcinoma: An In Silico and In Vitro Approach.

Purpose: Sorafenib is the sole FDA approved drug conventionally used for the treatment of advanced hepatocellular carcinoma (HCC). Despite of the beneficial use of sorafenib in the treatment of HCC, multidrug resistance still remains a challenge. HCC is inherently known as chemotherapy resistant tumor due to P-glycoprotein (P-gp)-mediated multidrug resistance. Methods: We studied the interaction energy of kaempferol with human multidrug resistance protein-1 (RCSB PDB ID: 2CBZ) using in silico method with the help of BIOVIA Discovery Studio. HepG2 and N1S1 liver cancer cell lines were treated in suitable cell culture media to evaluate the efficacy of kaempferol in chemo-sensitizing liver cancer cells towards the effect of sorafenib. Cell viability study was performed by MTT assay. Results: In silico analysis of kaempferol showed best docking score of 23.14 with Human Multi Drug Resistant Protein-1 (RCSB PDB ID: 2CBZ) compared with positive control verapamil. Inin-vitro condition, combination of sub-toxic concentrations of both kaempferol and sorafenib produced 50% cytotoxicity with concentration of 2.5 µM each which indicates that kaempferol has the ability to reverse the MDR by decreasing the over-expression of P-gp. Conclusion: Kaempferol is able to sensitize the HepG2 and N1S1 against the sub-toxic concentration of sorafenib. Hence, we consider that the efficacy of sorafenib chemotherapy can be enhanced by the significant approach of combining the sub-toxic concentrations of sorafenib with kaempferol. Thus, kaempferol can be used as a better candidate molecule along with sorafenib for enhancing its efficacy, if validated through preclinical studies.

Silence of cytoskeleton-associated protein 2 represses cell proliferation and migration and promotes apoptosis in liver cancer cell lines.

To investigate the roles of cytoskeleton-associated protein 2 (CKAP2) in proliferation, apoptosis, and migration in liver cancer cells and the potential mechanisms. Human normal hepatocyte L02 and liver cancer cell lines HepG2, Huh7, and SMMC-7721 were cultured. The CKAP2 expression was detected by real-time PCR and Western blotting. HepG2 cells were randomly divided into a control group, a negative control (NC) group, and a CKAP2 silencing (siCKAP2) group. CCK-8 and BrdU assays were used to evaluate cell viability and proliferation, respectively. Transwell assay was employed to determine cell migration and invasion. The protein levels of cleaved-caspase 3, Bax, E-cadherin, N-cadherin, Vimentin, phosphorylated Janus kinase 2 (p-JAK2), and phosphorylated signal transducer and activator of transcription 3 (p-STAT3) were determined by Western blotting. Compared with normal hepatocyte L02, CKAP2 was highly expressed in liver cancer cell lines HepG2, Huh7, and SMMC-7721 (all P<0.05). Compared with the NC group, cell viability and proliferation rate of the siCKAP2 group were decreased (both P<0.05). The apoptotic rate, protein expression of cleaved-caspase 3 and Bax in the siCKAP2 group were significantly higher than those in the NC group (all P<0.05). Compared with the NC group, cell migration and invasion rates of the siCKAP2 group were significantly attenuated (both P<0.05). Compared with the NC group, E-cadherin protein expression in siCKAP2 group was increased, while protein expression levels of Vimentin, N-cadherin, p-JAK2, and p-STAT3 were decreased (all P<0.05). CKAP2 gene silence inhibits proliferation, migration, and invasion, and promotes apoptosis in liver cancer cells, while JAK2/STAT3 signaling pathway may be involved in these processes.