Human Hepatoma Cancer Cell Lines Research Articles

Synthesis of substituted terpyridine nickel nitrate complexes and their inhibitory selectivity against cancer cell lines

Six terpyridine‑nickel complexes 1–6 were formed by the coordination of 4′-(4-R-phenyl)-2,2′:6′,2″-terpyridine (R = hydroxyl (L1), methoxyl (L2), methylsulfonyl (L3), fluoro (L4), bromo (L5), iodo (L6)) derivatives to nickel nitrate. The compositions and structures of these complexes were analyzed by Fourier Transform infrared spectroscopy (FT-IR), elemental analyses, electrospray ionization mass spectra (ESI-MS), solid-state ultraviolet-visible (UV–Vis) spectroscopy, and single crystal X-ray diffraction (1, 2 and 4) studies. In vitro anticancer cell proliferation experiments against SiHa (human cervical squamous cancer cell line) cells, Bel-7402 (human hepatoma cancer cell line), Eca-109 (human esophageal cancer cell line) and HL-7702 (human normal hepatocyte cell line) indicate that they have more excellent anti-proliferation effects than the cis-platin against Siha cells, Bel-7402 cells and Eca-109 cells. Especially, complex 5 showed a rather outstanding inhibitory effect against the SiHa cell line and was less toxic than the other compounds to the HL-7702 cell line, implying an obvious specific inhibitory effect. Therefore, complex 5 has the potential value to be developed as an anticancer cell-specific drug against human cervical squamous carcinoma. Molecular docking simulation, UV–vis absorption spectroscopy and circular dichroism experiments show that they prefer to bind to DNA part in an embedded binding manner.

Fe(II) complexes of 2,2':6',2''-terpyridine ligands functionalized with substituted-phenyl groups: synthesis, crystal structures and anticancer potential.

With the aim of developing potential anticancer drug candidates, a series of Fe(II) complexes were synthesized using nine 2,2':6',2''-terpyridine ligands functionalized with substituted-phenyl groups, and their biological activities were systematically investigated. Their bis-terpyridine sandwich-like structures were determined by single crystal X-ray crystallography. In vitro antiproliferative experiments based on three human cancer cell lines, including human hepatoma cancer cell line (Bel-7402), human esophageal cancer cell line (Eca-109), and human cervical squamous cancer cell line (SiHa), indicate the high antiproliferation activities of these complexes compared with commercial cisplatin. And their toxicity to normal cells was estimated based on human normal hepatocyte (HL-7702) cell line. In particular, when the phenyl in terpyridine ligand was modified by a carboxyl group, the corresponding complex 3 exhibited much higher antiproliferation to cancer Bel-7402 cells (IC50 = 3.653 μmol L-1) than cisplatin and low toxicity to normal HL-7702 cells (IC50 = 99.92 μmol L-1), implying a significant selectivity for 3 in killing hepatoma cancer cells. Combined with the fact that iron element is more accessible than platin, this series of Fe(II) complexes comprises potential candidates for anticancer drugs with specific inhibition of hepatoma cancer. UV titration experiments and circular dichroism (CD) showed a strong binding affinity between these nine complexes and CT-DNA. However, molecular docking simulation revealed the competitive binding of DNA and protein to these complexes. Further, the interactions between these complexes and bovine serum albumin (BSA) have been studied by fluorescence titration and CD spectroscopy.

In Vitro Antioxidant, Antiinflammation, and Anticancer Activities and Anthraquinone Content from Rumex crispus Root Extract and Fractions.

Rumex crispus is a perennial plant that grows in humid environments across Korea. Its roots are used in traditional Korean medicine to treat several diseases, including diseases of the spleen and skin and several inflammatory pathologies. In this study, different solvent fractions (n-hexane, dichloromethane, ethyl acetate, n-butanol, and aqueous fractions) from an ethanol extract of R. crispus roots were evaluated for the presence and composition of anthraquinone compounds and antioxidants by checking for such things as free radical scavenging activity, and electron and proton atom donating ability. In addition, anti-inflammatory activity was measured by NO scavenging activity and inflammatory cytokine production; furthermore, anti-cancer activity was measured by apoptosis-inducing ability. Polyphenolic and flavonoid compounds were shown to be abundant in the dichloromethane and ethyl acetate fractions, which also exhibited strong antioxidant activity, including free radical scavenging and positive results in FRAP, TEAC, and ORAC assays. HPLC analysis revealed that the dichloromethane fractions had higher anthraquinone contents than the other fractions; the major anthraquinone compounds included chrysophanol, emodin, and physcione. In addition, results of the anti-inflammatory assays showed that the ethyl acetate fraction showed appreciable reductions in the levels of nitric oxide and inflammatory cytokines (TNF-α, IL-1β, and IL-6) in Raw 264.7 cells. Furthermore, the anthraquinone-rich dichloromethane fraction displayed the highest anticancer activity when evaluated in a human hepatoma cancer cell line (HepG2), in which it induced increased apoptosis mediated by p53 and caspase activation.

Synthesis and evaluation of the antiproliferative efficacy of BRM270 phytocomposite nanoparticles against human hepatoma cancer cell lines

BRM270 is the most leading phytochemical extract that possesses potent anticancer properties. A major challenge associated with this drug is its low bioavailability and thus requires high dosages for cancer treatment. Here, we report the novel nano-synthesis of phyto-composite, BRM270 for the first time by mechanical milling method with specific modifications for enhanced cytotoxicity against HepG2 human hepatoma cancer cells. Unlike free BRM270 and other phytomedicines, BRM270 nanoparticles (BRM270 NPs) are well-dispersed and small sized (23 to 70 nm) which is believed to greatly enhanced cellular uptake. Furthermore, the acidic tumor microenvironment attracts BRM270 NPs enhancing targeted therapy while leaving normal cells less affected. The comparative cytotoxicity analysis using MTT assay among the three treatment groups, such as free BRM270, BRM270 NPs, and doxorubicin demonstrated that BRM270 NPs induced greater cytotoxicity against HepG2 cells with an effective drug concentration of 12 μg/ml. From FACS analysis, we observed an apoptotic cell death of 44.4% at BRM270 NPs treated cells while only 12.5% found in the free BRM270 treated cells. Further, the comparative relative expression profiling of the candidate genes were showed significant (p < 0.05) down-regulation of IL6, BCL2, p53, and MMP9 in the BRM270 NPs treated cells, compared to the free BRM270 and doxorubicin. Indeed, the genes, CASPASE 9 and BAX have shown significant (p < 0.05) upregulation in cells treated with BRM270 NPs as compared to counter treatment groups. The investigation of the signal pathways and protein-protein network associations were also carried out to elucidate the functional insights underlying anti-cancer potential of BRM270 NPs in HepG2 cells. Taken together, our findings demonstrated that these uniquely engineered BRM270 NPs effectively enter into the cancer cells due to its acidic microenvironment thereby inducing apoptosis and regulate the cell-proliferation in-vitro at extremely low dosages.

Evolution of a Strategy for the Enantioselective Total Synthesis of (+)-Psiguadial B.

(+)-Psiguadial B is a diformyl phloroglucinol meroterpenoid that exhibits antiproliferative activity against the HepG2 human hepatoma cancer cell line. This full account details the evolution of a strategy that culminated in the first enantioselective total synthesis of (+)-psiguadial B. A key feature of the synthesis is the construction of the trans-cyclobutane motif by a Wolff rearrangement with in situ catalytic, asymmetric trapping of the ketene. An investigation of the substrate scope of this method to prepare enantioenriched 8-aminoquinolinamides is disclosed. Three routes toward (+)-psiguadial B were evaluated that featured the following key steps: (1) an ortho-quinone methide hetero-Diels-Alder cycloaddition to prepare the chroman framework, (2) a Prins cyclization to form the bridging bicyclo[4.3.1]decane system, and (3) a modified Norrish-Yang cyclization to generate the chroman. Ultimately, the successful strategy employed a ring-closing metathesis to form the seven-membered ring and an intramolecular O-arylation reaction to complete the polycyclic framework of the natural product.

English

In this study, the ability of Tinospora crispa aqueous extract (TCAE) to regulate cholesterol metabolism in human hepatoma cancer cell line (Hep G2) was determined. Cytotoxic study was performed by exposing hepatoma cell (Hep G2) towards TCAE with concentration ranging from 0.002 to 20 mg/ml for 24 h at 37&deg;C and with 5% CO2 atmosphere. Result revealed that TCAE was not toxic to the cell. The ability of TCAE to reduce cholesterol in cell culture experiment was carried out by pre-treating Hep G2 with selected concentrations of TCAE (10, 5, 2.5, 1.25 and 0.625 mg/ml) in 6-well plate before the cell was exposed to low density lipoprotein (LDL). The concentration of apolipoprotein A1 (Apo A1), lecithin-cholesterol acyltransferase (LCAT), low density lipoprotein receptor (LDLR), scavenger receptor B1 (SRB1) and hepatic Lipase (HL) which involve in reverse cholesterol transport (RCT) pathway were determined from the 6-well plate medium. The direct pathway of cholesterol synthesis was performed according to the instruction provided in HMG-CoA Reductase Assay Kit manuals. The results showed that TCAE significantly increase (p&lt;0.05) the concentration of Apo A1, LCAT, LDLR, SRB-1 and HL. The efficacy of these activities is appreciably good when compared with standard drug simvastatin. However, TCAE showed moderate effect in controlling mevalonate pathway. These findings suggested that TCAE has the potential to reduce cholesterol metabolism in Hep G2 cancer cell lines and the pathway of TCAE action possibly more on RCT. &nbsp; Key words: Tinospora crispa, cholesterol metabolism, reverse cholesterol transport, cytotoxic, Hep G2.

Preparation and use of combustion derived Bi2O3 for the generation of novel heterocycles via Suzuki-Coupling Reactions: potential application as anti-cancer agents

Bismuth oxide was synthesized via simple, rapid and energy efficient solution combustion synthesis (SCS) by employing sucrose as a fuel. This SCS-Bi2O3 was characterized by analytical techniques such as PXRD, SEM, EDX, UV-Visible and BET surface area measurement. Using the prepared SCS-Bi2O3, several classes of heterocyclic compounds were synthesized in good yields via Suzuki-coupling reaction in aqueous medium. Interestingly, the recovered SCS-Bi2O3 could be reused three times without a significant loss of catalytic activity. Further, the synthesized compounds were tested for cytotoxic activity against a human hepatoma cancer cell line (HepG2). Among these, compound 3n effectively inhibited the proliferation of these cells with an IC50 value of 8.4 μM. Thus, this paper describes the preparation of highly effective Bi2O3, which can be used for synthesizing various classes of heterocycles, including those having anti-cancer property.

Effect of silencing RRM2 by specific small interference RNA on biological behaviors of human hepatoma cancer cell line HepG2

Objective To investigate the effect of small interference RNA (siRNA) silencing RRM2 on the biological behaviors of human hepatoma cancer cell line HepG2 and and the molecular mechanisms.Methods RRM2 expression was knocked down in human hepatoma cancer cell line HepG2 by RRM2 siRNA.The expression level of RRM2 was detected in human hepatoma cancer cell line HepG2 and human normal liver cell line HL-7702 at mRNA and protein levels by real-time polymerase chain reaction (PCR) or Western blotting.The cell proliferation was measured by cell counting Kit-8 (CCK-8).The migration was observed by using Transwell cell migration system.The expression levels of B lymphocytes/leukemia-2 (bcl-2),bax and cytochrome C (Cyt-C) proteins were detected by using Western blotting.Ten male BALB/C nude mice were subcutaneously injected with siRNA-RRM2/HepG2,siRNA-NC/HepG2 or NEG/HepG2 cells respectively to study the growth of tumor in each group.Results siRNA-RRM2 could down-regulate the expression of RRM2 in HepG2 remarkably,and siRNA-RRM2 could inhibit both the proliferation [(41.20 ± 2.13) ％ vs (91.35 ± 10.22) ％] and migration ability of HepG2 cells (21.20 ±1.28 vs 132.50 ±7.88) as compared with the negative control group.siRNA-RRM2 could down-regulate the expression of bcl-2 and up-regulate the expression of bax and Cyt-C.Additionally,the volume of xenograft tumors in siRNA-RRM2 group was decreased significantly as compared with control group.The volume of xenograft tumors in siRNA-RRM2 group was (171.13 ± 28.12) mm3 and that in siRNA-NC group was (1487.41 ± 168.20) mm3 at 8th week.Conclusion siRNA-RRM2 can down-regulate the expression of RRM2 in the HepG2 cells and may play an important role in the regulation of biological behaviors of HepG2 cells,and also inhibit the tumor growth of nude mice model. Key words: RNA interference ; RRM2 gene ; Carcinoma,hepatocellular

Antiproliferative potential of astaxanthin-rich alga Haematococcus pluvialis Flotow on human hepatic cancer (HepG2) cell line

To investigate the efficacy of astaxanthin from unicellular green alga, Haematococcus pluvialis inhibited the cell growth and trigger the apoptosis in human hepatoma cancer cell line (HepG2) in a dose and time dependent manner. Cell viability was analyzed by 3-(4, 5-dimethylthiazol2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. The different concentration of astaxanthin viz. 5, 10, 15, 20 and 25μg/mL to incubate the cell lines at different time course. The lactate dehydrogenase leakage (LDH) content and reduced glutathione (GSH) levels were also estimated. The depletion of GSH may be involved in the induction of apoptosis of the cancer cells. The apoptosis was evaluated with Annexin-V and Propidium iodide under fluorescent microscopy study. Moreover, the efficacy of astaxanthin 25μg/mL significantly inhibits the hepatoma cancer cell growth and induces cell apoptosis. In addition, immunofluorescence staining and flow cytometry studies were revealed that further evidence to confirm that the astaxanthin 15 and 25μg/mL to trigger the apoptosis 42.55% and 58.55% for 24h, respectively. Taken together, the present study suggests that H. pluvialis astaxanthin may protect from liver cancer.

Anti-proliferative effects of cinnamaldehyde on human hepatoma cell lines

Abstract Cinnamaldehyde has been shown to exert diverse health benefits on humans. We investigated the molecular effects of cinnamaldehyde on the growth regulation mechanism and the Janus kinase–signal transducers and activators of the transcription (JAK–STAT) signalling pathway in human hepatoma cells. We found that cinnamaldehyde caused inhibition of cellular mitogenesis, partly by promoting apoptosis in two human hepatoma cancer cell lines – HepG2 and Hep3B. Amplified changes in caspase 3 activity, Bcl-2 protein expression, and mitochondrial cytochrome c release were displayed in cinnamaldehyde treatments in these cells. The ability of cinnamaldehyde to induce growth arrest was also verified by the observation that it significantly decreased the protein levels of cyclin D1 and proliferative cell nuclear antigen (PCNA) but increased the protein levels of p27Kip1 and p21Waf1/Cip1. Especially, the JAK2–STAT3/STAT5 signalling pathway was markedly blocked by cinnamaldehyde in human hepatoma cells. Suppressor of cytokine signalling 1 (SOCS1) and SOCS3 expressions were slightly activated by cinnamaldehyde. The specific JAK2 inhibitor, AG490, significantly prevented the phosphorylation of STAT3 and STAT5 proteins and enhanced the effect of cinnamaldehyde-inhibited cellular mitogenesis. Hence, these results suggest that cinnamaldehyde has a potent inhibitory effect against human hepatoma cell growth, and the JAK2–STAT3/STAT5 and that the apoptotic pathways may be important targets of cinnamaldehyde.

Novel Pd(ii)–salen complexes showing high in vitro anti-proliferative effects against human hepatoma cancer by modulating specific regulatory genes

We have reported the synthesis of a novel salen ligand and its mononuclear Pd-salen complexes derived from 2-{[2-hydroxy-3-{[(E)-(2-hydroxyphenyl)methylidene]amino}propyl)imino]methyl}phenol. The newly synthesized and isolated Pd(II) complexes have been identified and fully characterized by various physico-chemical studies viz., elemental analyses, IR, UV-Vis, (1)H, (13)C NMR spectroscopy, electron spray ionization mass spectrometry (ESI-MS) and TGA/DTA studies. The molecular structure of the salen ligand has been ascertained by single-crystal XRD and it is coordinated to Pd(II) ion through two nitrogen and two oxygen atoms. The UV-Vis data clearly suggest a square-planar environment around both the Pd(II) ions. The DNA binding studies of the synthesized compounds has been investigated by electron spectroscopy and fluorescence measurements. The results suggest that Pd(II) complexes bind to DNA strongly as compared to the free ligand. The free salen ligand and its Pd(II) complexes have also been tested against human hepatoma cancer cell line (Huh7) and results manifested exceptional anti-proliferative effects of the Pd(II) complexes. The anti-proliferative activity of Pd(II) complexes has been modulated by specific regulatory genes.

Perifosine induces cell cycle arrest and apoptosis in human hepatocellular carcinoma cell lines by blockade of Akt phosphorylation

Hepatocellular carcinoma (HCC) is one of the most common solid cancers, representing the third cause of cancer-related death among cirrhotic patients. Treatment of advanced HCC has become a very active area of research. Perifosine, a new synthetic alkylphospholipid Akt inhibitor, has shown anti-tumor activity by inhibition of Akt phosphorylation. In this study, the effect of perifosine on the cell proliferation and apoptosis in hepatoma cells has been investigated. Cell growth inhibition was detected by MTT assay, cell cycle was analyzed by flow cytometry, AnnexinV-FITC apoptosis detection kit was used to detect cell apoptosis, and protein expression was examined by Western blotting analysis. Our present studies showed that Akt phosphorylation was inhibited by perifosine in HepG2 and Bel-7402 human hepatocellular carcinoma cells. Perifosine inhibited the growth of HepG2 cells and Bel-7402 cells in a dose-dependent manner, and arrested cell cycle progression at the G(2) phase. Apoptosis induction became more effective with increasing perifosine concentration. The caspase cascade and its downstream effectors, Poly (ADP-ribose) polymerase (PARP), were also activated simultaneously upon perifosine treatment. The proapoptotic effect of perifosine was in part depending on regulation of the phosphorylation level of ERK and JNK. Perifosine cotreatment substantially increased cytotoxic effects of cisplatin in HepG2 cells. Down-regulating the expression of Bcl-2 and up-regulating the level of Bax may be the potential mechanism for this synergistic effect. Our findings suggest that the small molecule Akt inhibitor perifosine shows substantial anti-tumor activity in human hepatoma cancer cell lines, and is a good candidate for treatment combinations with classical cytostatic compounds in hepatocellular carcinoma.

Butein induces G 2/M phase arrest and apoptosis in human hepatoma cancer cells through ROS generation

We investigated the molecular effects of 3,4,2′,4′-tetrahydroxychalcone (butein) treatment in two human hepatoma cancer cell lines–HepG2 and Hep3B. Butein treatment inhibited cancer cell growth by inducing G 2/M phase arrest and apoptosis. Butein-induced G 2/M phase arrest was associated with increased ATM, Chk1, and Chk2 phosphorylations and reduced cdc25C levels. Additionally, butein treatment enhanced inactivated phospho-Cdc2 levels, reduced Cdc2 kinase activity, and generated reactive oxygen species (ROS) that was accompanied by JNK activation. The extent of butein-induced G 2/M phase arrest significantly decreased following pretreatment with N-acetyl- l-cysteine or glutathione and following JNK phosphorylation reduction by SP600125. Both N-acetyl- l-cysteine and glutathione also decreased butein-mediated apoptosis. Taken together, these results imply a critical role of ROS and JNK in the anticancer effects of butein.

Functional characterization of interleukin-15 gene transduction into the human natural killer cell line NKL

Genetic modification of natural killer (NK) cells is a potential approach to gene-based immunotherapy of cancer. We created human interleukin-15 (hIL-15) gene-modified NKL cells and investigated their functional characterization in vitro. A recombinant vector (pcDNA3-IL15) or control vector (pcDNA3) was transferred into NKL cells by an electroporation method. Standard reverse transcriptase-polymerase chain reaction (RT-PCR), flow cytometry and MTT methods were performed for NK cell proliferation, apoptosis, cytotoxicity assays and gene expression tests. Compared with parental NKL cells, hIL-15 gene modification promoted NK cell proliferation at low doses of IL-2 and inhibited cell apoptosis, which was associated with the up-regulation of anti-apoptosis genes Bcl-2, Bcl-xl and Mcl-1 as well as the down-regulation of apoptosis genes Bim and Noxa. Moreover, the anti-tumor activity of hIL-15 gene-transduced NKL cells against human hepatoma cancer cell line HepG2, H7402 and PLC/PRF-5 cells was enhanced, at least partly, through increasing expression of cytotoxicity-associated genes, including interferon (IFN)-gamma, perforin and FasL. The hIL-15 genetic modification could improve the proliferation, anti-apoptosis and natural cytotoxicity of NKL cells against hepatocarcinoma cells. These data suggest that hIL-15 gene-modified NKL cells could be useful for clinical cancer immunotherapy in the future.