Hepatocellular Cell Line Research Articles

In vitro cytotoxicity analysis of Zizyphus spina-christi stem bark extract on human cancer cell lines.

Zizyphus spina-christi (Rhamnaceae family) is an edible plant used in folk medicine. Therefore, it is of interest to report the cytotoxic effects of Z. spina-christi bark crude extract on human cell lines. Crude ethanol extract of Z. spina-christi bark was fractionated with increasing polarity (diethyl ether, chloroform, ethyl acetate and butanol fractions). The fractions were examined for their cytotoxicity against human colon cancer (HCT-116 and CACO-2), cervical cancer (HeLa and HEp-2), lung carcinoma (A-549), hepatocellular carcinoma (HepG-2), breast cancer (MCF-7) and prostate cancer (PC-3) cell lines using viability assay. Diethyl ether fraction of Z. spina-christi showed the highest cytotoxic effects among the four extracts of Z. spina-christi. The IC50 of diethyl ether fraction was 7.14, 11.2, 11.6, 15.4, 39.8, 42.2, 84.2 and 153.8 µg/ml on HepG-2, A-549, CACO-2, HCT-116, MCF-7, PC-3, HeLa, and HEp-2 cell lines, respectively. Data shows that the diethyl ether fraction of Z. spina-christi showed effective cytotoxic effects in colon, lung and hepatocellular cancer cell lines.

Engineered adeno-associated virus 3 vector with reduced reactivity to serum antibodies

The natural serotypes of adeno-associated virus (AAV) or their variants, such as AAV8 and AAV5, are commonly used as vectors in the clinical programs for liver-targeted gene therapy. While AAV8 vectors are not highly efficient at targeting primary human hepatocytes, AAV3 vectors have recently demonstrated remarkable efficiency at targeting both human and non-human primate hepatocytes. However, the presence of high levels of neutralizing antibodies (NAbs) impedes transduction into hepatocytes, representing a major obstacle to the clinical application of AAV3 vectors. Herein, we engineered the viral capsid to reduce its reactivity with pre-existing NAbs, thereby enhancing the transduction efficiency. By introducing three substitutions (S472A, S587A, and N706A) on the surface loop of AAV3B capsid protein, we generated a triple mutant AAV3 (AAV.GT5) vector with less reactivity to anti-AAV capsid NAbs. While the transduction efficiency of AAV.GT5 into human hepatocellular cell lines was similar to those of parental AAV3B, it was 50-fold higher for hepatocytes derived from humanized mice compared to AAV8 vectors. Moreover, the AAV.GT5 vector yield was similar to those of the AAV2 and AAV3B vectors. Thus, high resistance to pre-existing NAbs makes AAV.GT5 a promising candidate for future liver-targeted gene therapy clinical trials.

Functional roles of GRP78 in hepatitis B virus infectivity and antigen secretion.

Viruses utilize cellular proteins to mediate their life cycle. However, the hepatitis B virus (HBV) life cycle is still mysterious and remains to be elucidated. Here, GRP78/BiP/HSPA5, a 78 kDa glucose-regulated protein, was identified as a preS2 interacting protein. Pulldown assay showed the interaction of glucose-regulated protein 78 (GRP78) with both the preS2 domain-containing large S and middle S proteins expressed in a human hepatocellular cell line. The immunofluorescence studies revealed that the preS2 colocalized with GRP78. Interestingly, it was found that preS2 specifically bound to the ATPase domain of GRP78. To understand how GRP78 plays a role in HBV infection, stably GRP78-expressing cells were established, which promoted HBV infectivity and replication. In contrast, knockdown of GRP78 changed the HBV antigen secretion but not the viral DNA amplification. Taken together, these results suggest that GRP78 should interact with preS2 via the ATPase domain and modulate both the HBV infectivity and HBV antigen secretion.

Retracted] Hypoxia‑induced mitochondrial translocation of DNM1L increases mitochondrial fission and triggers mPTP opening in HCC cells via activation of HK2.

Subsequently to the publication of the above paper, the authors have retrospectively realized that they used the human normal liver cell, line L‑02, for the experiments reported in this study instead of the intended hepatocellular cell line, Huh‑7. Consequently, the results and conclusions reported in this article must be considered to lack reliability. Therefore, the authors have requested that the article be retracted from the publication. The authors apologize to the Editor and to the readership for any inconvenience caused. [the original article was published in Oncology Reports 42: 1125‑1132, 2019; DOI: 10.3892/or.2019.7213].

Abstract PO-030: The harmala alkaloid harmine as a novel cancer cell radiosensitizer

Abstract Radiation therapy resistance remains a persistent clinical challenge in the treatment of multiple different cancer types. Better appreciation of novel molecular targets to improve radiation therapy efficacy is needed. TWIST1 is an epithelial-mesenchymal transcription factor whose expression is linked to therapy resistance, including radioresistance, in cancer. We have shown previously that the harmala alkaloid harmine degrades TWIST1 and possess anti-tumor activity. We hypothesized that harmine may radiosensitize cancer cells via downregulation of TWIST1. We examined the effects of harmine combined with radiation (RT) on two TWIST1 overexpressing tumor cell lines, A549 human non-small cell lung cancer (NSCLC) cells and the LMT mouse-derived hepatocellular cancer cell line (HCC), both with doxycycline-inducible Twist1 expression. We also used the LM mouse-derived HCC cell line without Twist1 expression as a control. In vitro assays for clonogenic survival, apoptosis, cell cycle distribution, γH2AX foci kinetics, and protein expression in pathways important for cancer cell survival and radioresistance were performed. Harmine alone exhibited anti-cancer activity irrespective of Twist1 expression as shown by decreased clonogenic survival. Harmine caused a supra-additive radiosensitization in all cancer cell lines with radiation enhancement ratios between 1.05 – 3.04. In vivo we are evaluating tumor growth delay and effects of combined harmine and RT treatment in a NSCLC hind-flank tumor growth model. Mechanistic elucidation of these results, studies on normal cells, and in vivo validation are ongoing. Citation Format: Caleb R.P. Smack, Audrey Lafargue, Christine Lam, Francesca A. Carrieri, Ismaeel Siddiqui, Hailun Wang, Phuoc T. Tran. The harmala alkaloid harmine as a novel cancer cell radiosensitizer [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-030.

IN VITRO CYTOTOXIC EVALUATION OF KARHEE WEED

Locally famous Karhee or Berre (Sorbaria tomentosa) exhibits medicinal value is large woody shrub indigenous to Pakistan. The present study examined the cytotoxic activities of S. tomentosa using methanolic extracts and fractions (n-hexane, dichloromethane, ethyl acetate and water) against three cáncer cell lines (lung A-549, hepatocellular HepG2and urinary bladder EI-138). Cytotoxic assays were carried out with five different concentrations (0.05, 0.01, 0.05, 0.1 and 0.5 mg/mL) of methanolic extract and its sub fraction through MTT assay. Results revealed n-hexane and ethyl acetate fraction being the most potent against all test cancer cell lines with higher IC50 values. Both fractions also exhibited the maximum reduction in the cell viability in dose dependent manner. Preliminary results suggest the promising anticancer potential of n-hexane and ethyl acetate S. tomentosa against lung A-549, hepatocellular HepG2and urinary bladder EI-138 cell lines. Further studies are required to know the mechanism(s) involved in the cell death.

LvHemB1, a novel cationic antimicrobial peptide derived from the hemocyanin of Litopenaeus vannamei, induces cancer cell death by targeting mitochondrial voltage-dependent anion channel 1.

Current cancer treatment regimens such as chemotherapy and traditional chemical drugs have adverse side effects including the appearance of drug-resistant tumor cells. For these reasons, it is imperative to find novel therapeutic agents that overcome these factors. To this end, we explored a cationic antimicrobial peptide derived from Litopenaeus vannamei hemocyanin (designated LvHemB1) that induces cancer cell death, but sparing normal cells. LvHemB1 inhibits the proliferation of human cervical (HeLa), esophageal (EC109), hepatocellular (HepG2), and bladder (EJ) cancer cell lines, but had no significant effect on normal liver cell lines (T-antigen-immortalized human liver epithelial (THLE-3) cells). In addition to its antiproliferative effects, LvHemB1 induced apoptosis, by permeating cells and targeting mitochondrial voltage-dependent anion channel 1 (VDAC1). Colocalization studies revealed the localization of LvHemB1 in mitochondria, while molecular docking and pull-down analyses confirmed LvHemB1-VDAC1 interaction. Moreover, LvHemB1 causes loss in mitochondrial membrane potential and increases levels of reactive oxygen species (ROS) and apoptotic proteins (caspase-9, caspase-3, and Bax (Bcl-2-associated X)), which results in mitochondrial-mediated apoptosis. Thus, peptide LvHemB1 has the potential of being used as an anticancer agent due to its antiproliferation effect and targeting to VDAC1 to cause mitochondrial dysfunction in cancer cells, as well as its ability to induce apoptosis by increasing ROS levels, and the expression of proapoptotic proteins.

Cytotoxic and Genotoxic Effects of Cyanobacterial and Algal Extracts-Microcystin and Retinoic Acid Content.

In the last decade, it has become evident that complex mixtures of cyanobacterial bioactive substances, simultaneously present in blooms, often exert adverse effects that are different from those of pure cyanotoxins, and awareness has been raised on the importance of studying complex mixtures and chemical interactions. We aimed to investigate cytotoxic and genotoxic effects of complex extracts from laboratory cultures of cyanobacterial species from different orders (Cylindrospermopsis raciborskii, Aphanizomenon gracile, Microcystis aeruginosa, M. viridis, M. ichtyoblabe, Planktothrix agardhii, Limnothrix redekei) and algae (Desmodesmus quadricauda), and examine possible relationships between the observed effects and toxin and retinoic acid (RA) content in the extracts. The cytotoxic and genotoxic effects of the extracts were studied in the human hepatocellular carcinoma HepG2 cell line, using the MTT assay, and the comet and cytokinesis-block micronucleus (cytome) assays, respectively. Liquid chromatography electrospray ionization mass spectrometry (LC/ESI-MS) was used to detect toxins (microcystins (MC-LR, MC-RR, MC-YR) and cylindrospermopsin) and RAs (ATRA and 9cis-RA) in the extracts. Six out of eight extracts were cytotoxic (0.04–2 mgDM/mL), and five induced DNA strand breaks at non-cytotoxic concentrations (0.2–2 mgDM/mL). The extracts with genotoxic activity also had the highest content of RAs and there was a linear association between RA content and genotoxicity, indicating their possible involvement; however further research is needed to identify and confirm the compounds involved and to elucidate possible genotoxic effects of RAs.

Effect of Valproic Acid on the Class I Histone Deacetylase 1, 2 and 3, Tumor Suppressor Genes p21WAF1/CIP1 and p53, and Intrinsic Mitochondrial Apoptotic Pathway, Pro- (Bax, Bak, and Bim) and anti- (Bcl-2, Bcl-xL, and Mcl-1) Apoptotic Genes Expression, Cell Viability, and Apoptosis Induction in Hepatocellular Carcinoma HepG2 Cell Line.

Hepatocellular carcinoma (HCC), Primary liver cancer, is the fifth most common cancer in men. Histone deacetylation causes chromatin condensation resulting in gene silencing and tumorigenesis. These enzymes have become a novel target for the treatment of cancer. Histone deacetylase inhibitors (HDACIs) can reactivate tumor suppressor genes (TSGs) by inhibition of histone deacetylases (HDACs) activity leads to apoptosis induction in cancer cells. Further, these compounds can induce apoptosis through the intrinsic/mitochondrial pathway. Previously, we reported the effect of valproic acid (VPA) and trichostatin A (TSA) on TSGs p21WAF1/CIP1 (p21), p27Kip1 (p27), and p57Kip2 (P57) and also HDAC1 in colon cancer. The present study was designed to investigate the effect of VPA on the class I histone deacetylase (HDAC) 1, 2 and 3, TSGs p21and p53, and intrinsic mitochondrial pathway, pro- (Bax, Bak, and Bim) and anti- (Bcl-2, Bcl-xL, and Mcl-1) apoptotic genes, viability, and apoptosis in HCC HepG2 cell line. The HepG2 cells were cultured and treated with VPA. To determine viability, apoptosis, and the relative expression level of the mentioned genes, MTT assay, cell apoptosis assay, and qRT-PCR were done respectively. VPA downregulated class I histone deacetylase (HDAC) 1, 2, and 3, Bcl-2, Bcl-xL, and Mcl-1 and upregulated p21, p53, Bax, Bak, and Bim resulting in apoptosis induction. VPA can induce apoptosis via activation of the intrinsic mitochondrial apoptotic pathway and also epigenetic reactivation of p21 and p53 through inhibition of class I HDAC 1, 2 and 3, activity.<br />.

Hemecitones A and B: two phenanthrenes with cytotoxicity from Hemerocallis fulva (L.) L

Two new phenanthrenes, Hemecitones A and B, were isolated from Hemerocallis fulva (L.) L. Their structures were determined by UV, IR, HR-ESI-MS, 1D and 2D NMR analysis. The in vitro cytotoxic activities of compounds 1 and 2 were studied against breast carcinoma MDA-MB-231, hepatocellular carcinoma HepG2 and lung carcinoma A549 cell lines. And compounds 1 and 2 exhibited anti-proliferative effects against these least one of the three types of cell lines with IC50 ranging from 12.57 ± 2.34 to 31.22 ± 1.36 μM.

Estradiol is significantly associated with prognosis in non-surgical liver cancer patients: from bench to bedside.

There are rarely systematic studies to analyze the prognostic factors among non-surgical liver cancer patients. Whether there is a gender difference in the survival of non-surgical liver cancer patients and what may cause this difference is still unclear.A total of 12,312 non-surgical liver cancer patients were enrolled in this study. Age, race, sex, grade, tumor TNM stage, marital status, tumor size, and histological type were independent risk factors in liver cancer and were confirmed in the validation cohort. Before menopause, females demonstrated a better mean survival probability than males (39.4±1.4 vs. 32.7±0.8 months, respectively; p<0.001), and continued in post-menopause. The results of differentially expressed genes (DEGs) and KEGG pathway analysis showed that there were significant differences in steroid hormone biosynthesis between male and female liver cancer patients. In vitro experiments revealed that estradiol inhibited the proliferation of hepatocellular cancer cell lines and increased apoptosis, but estrone exerted no effect.In conclusion, gender differences in prognosis among non-surgical liver cancer patients were confirmed and attributable primarily to estradiol.

The expression of CD44, CD90 and CD133 in response to cisplatin in hepatocellular cancer cells

Introduction. Cancer is a leading cause of mortality. Hepatocellular cancer is one of the malignancies associated with poor outcome and resistance to pharmacotherapy. Cancer stem cells (CSCs) contribute to resistance to therapy and hence lead to the treatment failure of tumors. Aim. This study aims to explore the expression of CSCs in response to cisplatin treatment in HepG2 hepatocellular cancer cell line. Material and methods. Cell proliferation test, CCK-8, was used to evaluate the cell proliferation following cisplatin treatment for 72 hours. The expressions of CSC markers CD44, CD90, and CD133 were assessed by flow cytometric analysis. Results. The results showed a dose-dependent decrease in cell proliferation and increased expression of CSC markers CD44 and CD90 in response to cisplatin. Conclusion. Understanding the roles of CSC markers may point to new targets and therapeutic strategies to predict and overcome cisplatin resistance.

Virucidal activity of oriental hornet Vespa orientalis venom against hepatitis C virus.

BackgroundHepatitis C virus (HCV) infection is a major worldwide health problem that can cause liver fibrosis and hepatocellular carcinoma (HCC). The clinical treatment of HCV infection mainly relies on the use of direct-acting antivirals (DAAs) that are usually expensive and have side effects. Therefore, achieving the discovery of more successful agents is always urgent. In this context, antiviral compounds that inhibit viral infections and disease progression with important therapeutic activities have been identified in animal venoms including arthropod toxins. This indicates that arthropod venoms represent a good natural source of promising candidates for new antivirals. MethodsThe antiviral activity of the wasp venom (WV), isolated from the Oriental hornet (Vespa orientalis), was assessed using cell culture technique with human hepatocellular carcinoma-derived cell line (Huh7it-1) and the recombinant strain of HCV genotype 2a (JFH1). ResultsThe results revealed that WV inhibited HCV infectivity with 50% inhibitory concentration (IC50) of 10 ng/mL, while the 50% cytotoxic concentration (CC50) was 11,000 ng/mL. Time of addition experiment showed that the WV blocked HCV attachment/entry to the cells probably through virucidal effect. On the other hand, the venom showed no inhibitory effect on HCV replication. ConclusionWV can inhibit the entry stage of HCV infection at non-cytotoxic concentrations. Therefore, it could be considered a potential candidate for characterization of natural anti-HCV agents targeting the entry step.

Phytochemical Biological and Cytotoxic Activities of Calothamnus quadrifidus Leaves Cultivated in Egypt

The hydro distilled essential oil obtained from Calothamnus quadrifidus leaves was analyzed by gas liquid chromatography-mass spectroscopy. Quantification of the oil constituents was carried out using the gas liquid chromatography-flame ionization detection device where 1,8-cineole was quantified as 525.62 μg/ml. The oil showed potential antioxidant activity when tested by 1,1-diphenyl-2-picrylhydrazine radical scavenging assay where the concentration required to scavenge 1,1-diphenyl-2-picrylhydrazine by 50 %=37.3 μg. The cytotoxic activity of the essential oil was evaluated using 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide assay compared with 1,8-cineole and doxorubicin. Calothamnus quadrifidus leaves essential oil cytotoxic activity was significant against human lung cell line A-549 (2.37 μg), human colon cell line HCT-116 (2.39 μg), human hepato-cellular cell line HepG-2 (2.55 μg) and intestinal epithelium carcinoma Caco-2 (5.80 μg) cell lines, respectively. The oil cytotoxicity exceeded 1,8-cineole except for intestinal epithelium carcinoma Caco-2. Additionally, the antimicrobial activity was evaluated using agar disc diffusion and minimum inhibitory concentration where the oil exhibited the best activity against Penicillium italicum and Geotrichum candidum with minimum inhibitory concentration=0.49 and 0.98 μg, respectively. Structure elucidation of the pure compounds isolated from Calothamnus quadrifidus leaves was done according to their chromatographic behavior, chemical and spectroscopic data. These compounds were isolated for the first time from Calothamnus quadrifidus leaves and identified as gallic acid, gallic acid methyl ester, 7,4`-dimethoxy-apigenin, 6,7-dimethoxy-apigenin, (Bis[3(2-methyl propan-1- one)-2, 6 dihydroxy-4- methoxy phenyl] methane, quercetin 3-O-beta-D-arbinopyranoside and quercetin 3-O-beta-D-4C1-xylopyranoside. In addition to the molecular docking of novel compound 3 was done in peroxiredoxins and the binding mode was found. Conformation 2 was found to be the most stable and may be responsible for the antioxidant effect as it showed the best free binding energy and affinity when compared to conformations.

Anticancer, antimicrobial, and dye degradation activity of biosynthesised silver nanoparticle using Artemisia kopetdaghensis

Biosynthesis of nanoparticles (NPs) is gaining attention due to the presence of functional plant macromolecules that benefit from NPs’ synthesis and also due to their exceptional bioactive compounds. Artemisia kopetdaghensis extract acted as a reducing/capping agent. Characterisation of greener-synthesised AgNPs (Ak-AgNPs) was performed by various techniques, such as Ultraviolet (UV)−visible spectrophotometry, TEM, Fourier transform infrared, and XRD. Ak-AgNPs indicated great bactericidal properties in terms of zone of inhibition (ZI) against all of the pathogenic bacteria (i.e. Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus pyogenes, Staphylococcus epidermidis, and Staphylococcus aureus). The maximum ZI of Ak-AgNPs was 22.3 and 17.8 mm against K. pneumoniae and S. aureus, respectively. Anti-proliferative activity of Ak-AgNPs on human hepatocellular cancer cell line (HepG2) using the MTT test showed dosage-dependent inhibition activity of Ak-AgNPs against HepG2 cancer cells. Also, green-synthesised Ak-AgNPs showed catalytic properties under UV-light in organic dye (methylene blue) degradation. This study revealed that the green-synthesised AgNPs using A. kopetdaghensis shoots extract had antibacterial and catalytic activities. Also, Ak-AgNPs had anti-proliferative potential against human liver cancer cell lines. The green-synthesised Ak-AgNPs have the potential to be exploited in anti-bacterial, anticancer, and biocatalyst technologies.

Optimization of the spherical integrity for sustained-release alginate microcarriers-encapsulated doxorubicin by the Taguchi method

This study aimed to develop biodegradable calcium alginate microcarriers with uniform particle size and spherical integrity for sustained-release targeting transarterial chemoembolization. To determine related parameters including the ratio of cross-linking volume (sodium alginate: CaCl2), concentrations of sodium alginate and CaCl2 solutions, collection distance, flow rate, stirring speed, syringe needle diameter and hardening time to fabricate the microcarriers, the Taguchi method was applied. Using different conditions, a total of 18 groups were prepared. The average size of microspheres from different groups was estimated as ~ 2 mm (range 1.1 to 1.6 mm). Signal-to-noise ratio analysis showed the optimal spherical integrity (F1) achieved when the above parameters were designed as 0.1, 2.5 wt%, 6 wt%, 8 cm, 30 mL/h, 150 rpm, 0.25 mm and 2 h, respectively. The best (F1), middle (F2) and worst (F3) groups were used for further experiments. Fourier-transform infrared spectroscopy spectrum showed that F1, F2 and F3 conformations were distinct from original sodium alginate. Drug-loaded calcium alginate microcarriers demonstrated rougher surfaces compared to microspheres without drug under transmission electron microscopy. Compared to pH 7.4, swelling rates in PBS were decreased at pH 6.5. Encapsulation and loaded efficiencies of the Dox-loaded microcarriers were estimated as ~ 40.617% and ~ 3.517%. In vitro experiments indicated that the F1 Dox-loaded microcarriers provide a well sustained-release efficacy for about two weeks at 37 °C in PBS. Treatments of calcium alginate microcarriers without the Dox in two distinct hepatocellular carcinoma-derived cell lines, Huh-7 and Hep-3B, indicated that these microcarriers were non-toxic. The Dox-loaded microcarriers displayed sustained-release capacity and reduced cell viabilities to ~ 30% in both cell lines on Day 12.

Synthesis, in vitro biological activity, hydrolytic stability and docking of new analogs of BIM-23052 containing halogenated amino acids.

One of the potent somatostatin analogs, BIM-23052 (DC-23-99) D-Phe-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-NH2, has established in vitro growth hormone inhibitory activity in nM concentrations. It is also characterized by high affinity to some somatostatin receptors which are largely distributed in the cell membranes of many tumor cells. Herein, we report the synthesis of a series of analogs of BIM-23052 containing halogenated Phe residues using standard solid-phase peptide method Fmoc/OtBu-strategy. The cytotoxic effects of the compounds were tested in vitro against two human tumor cell lines-breast cancer cell line and hepatocellular cancer cell line, as well as on human non-tumorigenic epithelial cell line. Analogs containing fluoro-phenylalanines are cytotoxic in μM range, as the analog containing Phe (2-F) showed better selectivity against human hepatocellular cancer cell line. The presented study also reveals that accumulation of halogenated Phe residues does not increase the cytotoxicity according to tested cell lines. The calculated selective index reveals different mechanisms of antitumor activity of the parent compound BIM-23052 and target halogenated analogs for examined breast tumor cell lines. All peptides tested have high antitumor activity against the HepG2 cell line (IC50 ≈ 100μM and SI > 5) compared to breast cells. This is probably due to the high permeability of the cell membrane and the higher metabolic activity of hepatocytes. In silico docking studies confirmed that all obtained analogs bind well with the somatostatin receptors with preference to ssrt3 and ssrt5. All target compounds showed high hydrolytic stability at acid and neutral pH, which mimic physiological condition in stomach and human plasma.

GDF11 restricts aberrant lipogenesis and changes in mitochondrial structure and function in human hepatocellular carcinoma cells.

Growth differentiation factor 11 (GDF11) has been characterized as a key regulator of differentiation in cells that retain stemness features. Recently, it has been reported that GDF11 exerts tumor-suppressive properties in hepatocellular carcinoma cells, decreasing clonogenicity, proliferation, spheroid formation, and cellular function, all associated witha decrement in stemness features, resulting in mesenchymal to epithelial transition and loss of aggressiveness. The aim of the present work was to investigate the mechanism associated with the tumor-suppressive properties displayed by GDF11 in liver cancer cells. Hepatocellular carcinoma-derived cell lines were exposed to GDF11 (50 ng/ml), RNA-seq analysis in Huh7 cell line revealed that GDF11 exerted profound transcriptomic impact, which involved regulation of cholesterol metabolic process, steroid metabolic process as well as key signaling pathways, resembling endoplasmic reticulum-related functions. Cholesterol and triglycerides determination in Huh7 and Hep3B cells treated with GDF11 exhibited a significant decrement in the content of these lipids. The mTOR signaling pathway was downregulated, and this was associated witha reduction in key proteins involved in the mevalonate pathway. In addition, real-time metabolism assessed by Seahorse technology showed abridged glycolysis as well as glycolytic capacity, closely related to an impaired oxygen consumption rate and decrement in adenosine triphosphate production. Finally, transmission electron microscopy revealed mitochondrial abnormalities, such as cristae disarrangement, consistent with metabolic changes. Results provide evidence that GDF11 impairs cancer cell metabolism targeting lipid homeostasis, glycolysis, and mitochondria function and morphology.

Next-generation sequencing reveals alternative L-DOPA decarboxylase (DDC) splice variants bearing novel exons, in human hepatocellular and lung cancer cells.

The human L-DOPA decarboxylase (DDC) is an enzyme that displays a pivotal role in metabolic processes. It is implicated in various human disorders, including hepatocellular and lung cancer. Several splice variants of DDC have previously been described, most of which encode for protein isoforms of this enzyme. In the present study, we used next-generation sequencing (NGS) technology along with nested touchdown PCR and Sanger sequencing to identify new splice variants bearing novel exons of the DDC gene, in hepatocellular and lung cancer cell lines. Using an in-house-developed algorithm, we discovered seven novel DDC exons. Next, we determined the structure of ten novel DDC transcripts, three of which contain an open reading frame (ORF) and probably encode for three previously unknown protein isoforms of this enzyme. Future studies should focus on the elucidation of their role in cellular physiology and cancer pathobiology.

New Platinum (II) Ternary Complexes of Formamidine and Pyrophosphate: Synthesis, Characterization and DFT Calculations and In vitro Cytotoxicity.

Platinum (II) and platinum (IV) of pyrophosphate complexes have been prepared and characterized to discover their potential as antitumor drugs. This study was conducted to prepare and characterize new ternary platinum (II) complexes with formamidine and pyrophosphate as an antitumor candidate. The complexes have been characterized by mass, infrared, UV-Vis. spectroscopy, elemental analysis, magnetic susceptibility, thermal analyses, and theoretical calculations. They have been tested for their cytotoxicity, which was carried out using the fastcolorimetric assay for cellular growth and survival against MCF-7 (breast cancer cell line), HCT- 116 (colon carcinoma cell line), and HepG-2 (hepatocellular cancer cell line). All complexes are diamagnetic, and the electronic spectral data displayed the bands due to square planar Pt(II) complexes. The optimized complexes structures (1-4) indicated a distorted square planar geometry where O-Pt-O and N-Pt-N bond angles were 82.04°-96.44°, respectively. Results also show that all complexes are neutral, stable and non-hygroscopic and have noticeable cytotoxicity with IC50 (μM): 0.035-0.144 MCF-7(breast cancer cell line), 0.042-0.187 HCT-116 (colon carcinoma cell line), and 0.063-0.168 HepG-2 (hepatocellular cancer cell line). Moreover, the results show that the complex (4) has the best IC50 value. The complexes showed noticeable cytotoxicity and are considered as promising antitumor candidates for further applications.