Lung Epithelial Carcinoma Cells Research Articles

Anticancer effects of alloxanthoxyletin and fatty acids esters – In vitro study on cancer HTB-140 and A549 cells

Alloxanthoxyletin, a natural occurring pyranocoumarin isolated from a number of plant sources, such as family of Rutaceae, and its synthetic derivatives show cytotoxic and antitumor activities. In the present study new eleven esters of alloxanthoxyletin and fatty acids were synthesized and evaluated for their anticancer toxicity. The structures of the compounds were confirmed by Proton Nuclear Magnetic Resonance (1H NMR), Carbon-13 Nuclear Magnetic Resonance (13C NMR) and High Resolution Mass Spectrometry (HRMS) analyses.For all compounds 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cytotoxic effect on human melanoma cells (HTB-140), human epithelial lung carcinoma cells (A549) and human keratinocyte line (HaCaT). For the most active compounds (8–11) lactate dehydrogenase (LDH) assay to assess the level of cell damage as well as migration inhibition assay were performed. To explain the basic mechanism of cell death induction, the effect of derivatives 8–11 on early and late apoptosis in Annexin V-FITC/7-AAD flow cytometry analysis was investigated.The results indicate that human melanoma cells (HTB-140) and human epithelial lung carcinoma cells (A549) were more sensitive to new alloxanthoxyletin derivatives exposure compared to human keratinocytes (HaCaT). Both, the cytotoxicity and the migration tests showed a concentration-dependent inhibition of cell growth, although with a different degree of efficacy. Tested compounds induced apoptosis in cancer cells, however, derivatives 8, 9, 10 and 11 were found to be much more potent inducers of early apoptosis in HTB-140 cells than in A549 and HaCaT cells.To establish the potent mechanism of action of alloxanthoxyletin derivatives 8, 9, 10 and 11 on HaCaT, A549 and HTB-140 cells, the level of IL-6 was measured. Our results indicate, that tested compounds significantly decrease the release of IL-6 for all cancer cell lines.

Triazine- and Heptazine-Based Carbon Nitrides: Toxicity

Graphitic carbon nitride has attracted extensive interests recently because of its potential in biosensing, photocatalytic, and biomedical applications. Similar to graphene, it is a two-dimensional carbon and nitrogen-based nanomaterial with weak van der Waals forces between each layer. Carbon nitrides can have various structural moieties such as triazine and heptazine. Unlike graphene-substituted nanosheets, the toxicity of graphitic carbon nitrite is largely unknown. In respond to that, toxicological study was carried out to determine its toxicity toward human lung carcinoma epithelial cells (A549). Two formazan-based cell viability assays (water-soluble tetrazolium salt (WST-8) assay and methylthiazolyldiphenyltetrazolium bromide (MTT) assay) were utilized on the A549 cell line to derive the cytotoxicity profile. Both materials demonstrated a dose-dependent toxicological effect with triazine-based carbon nitrides being more cytotoxic than heptazine-based carbon nitrides. These findings are of great imp...

EmPis-1L, an Effective Antimicrobial Peptide Against the Antibiotic-Resistant VBNC State Cells of Pathogenic Bacteria.

The antibiotic-resistant viable but non-culturable (VBNC) pathogenic bacteria are considered as a new threat to public health. Antimicrobial peptides (AMPs), possessing bactericidal effects in a rapid membrane attacking mode, are supposed to be effective against bacteria entering the VBNC state. In the current study, the activity of grouper AMP piscidin killing the VBNC state cells of pathogenic bacteria Escherichia coli O157, Staphylococcus aureus, and Vibrio parahaemolyticus OS4 was studied. After entering the VBNC state, cells of E. coli O157, S. aureus, and V. parahaemolyticus OS4 developed resistance to the antibiotics Ampicillin and Kanamycin. Rather than truncated form of Malabar grouper piscidin 1 (EmPis-1S), full-length Malabar grouper piscidin 1 (EmPis-1L) showed strong activity to kill the above VBNC bacteria. The VBNC state cells (1 × 105CFU/mL) of the three species of bacteria could be totally lysed by 10μmol/L of EmPis-1L in 1h. The VBNC state cells of S. aureus were most susceptible to EmPis-1L, which killed the cells by 100% in 30min at the low concentration of 2.0μmol/L. In addition, EmPis-1L at the concentration of no more than 10μmol/L showed no observed toxicity to human lung carcinoma epithelial cells (A549) and mouse neuroblastoma cells (N2a). Accordingly, EmPis-1L could be a promisingly safe and efficient agent for eliminating the traditional antibiotic-resistant VBNC state cells of pathogenic bacteria, E. coli, S. aureus, and V. parahaemolyticus.

Regulation and biological role of the peptide/histidine transporter SLC15A3 in Toll-like receptor-mediated inflammatory responses in macrophage

The peptide/histidine transporter SLC15A3 is responsible for transporting histidine, certain dipeptide and peptidomimetics from inside the lysosome to cytosol. Previous studies have indicated that SLC15A3 transcripts are mainly expressed in the lymphatic system, however, its regulation and biological role in innate immune responses and inflammatory diseases are as yet unknown. In this study, mouse peritoneal macrophages (PMs), mouse bone marrow-derived macrophages (BMDMs), the human acute monocytic leukemia cell line THP-1 and the human lung epithelial carcinoma cell line A549 were used to investigate the regulation and biological role of SLC15A3 in TLR-mediated inflammatory responses. Our results showed that SLC15A3 was upregulated by TLR2, TLR4, TLR7 and TLR9 ligands in macrophages at both the mRNA and protein levels via activation of NF-κB (nuclear factor-kappa-B), MAPK (mitogen-activated protein kinase) and IRF3 (interferon regulatory factor 3). Furthermore, knockdown or overexpression of SLC15A3 influenced the TLR4-triggered expression of proinflammatory cytokines. A reporter gene assay showed that the SLC15A3 promotor contained potential NF-κB binding sites, which were reasonable for regulating SLC15A3 by TLR-activation through NF-κB signaling. Additionally, SLC15A3 expression was increased and positively related to inflammation in mice with bacterial peritonitis. The collective findings suggest that SLC15A3 is regulated by various TLRs, and that it plays an important role in regulating TLR4-mediated inflammatory responses.

An experimental investigation of a novel iron chelating protoporphyrin IX prodrug for the enhancement of photodynamic therapy.

ObjectivesNon‐melanoma skin cancers are the most frequently occurring type of cancer worldwide. They can be effectively treated using topical dermatological photodynamic therapy (PDT) employing protoporphyrin IX (PpIX) as the active photosensitising agent as long as the disease remains superficial. Novel iron chelating agents are being investigated to enhance the effectiveness and extend the applications of this treatment modality, as limiting free iron increases the accumulation of PpIX available for light activation and thus cell kill.MethodsHuman lung fibroblasts (MRC‐5) and epithelial squamous carcinoma (A431) cells were treated with PpIX precursors (aminolaevulinic acid [ALA] or methyl‐aminolevulinate [MAL]) with or without the separate hydroxypyridinone iron chelating agent (CP94) or alternatively, the new combined iron chelator and PpIX producing agent, AP2‐18. PpIX fluorescence was monitored hourly for 6 hours prior to irradiation. PDT effectiveness was then assessed the following day using the lactate dehydrogenase and neutral red assays.ResultsGenerally, iron chelation achieved via CP94 or AP2‐18 administration significantly increased PpIX fluorescence. ALA was more effective as a PpIX‐prodrug than MAL in A431 cells, corresponding with the lower PpIX accumulation observed with the latter congener in this cell type. Addition of either iron chelating agent consistently increased PpIX accumulation but did not always convey an extra beneficial effect on PpIX‐PDT cell kill when using the already highly effective higher dose of ALA. However, these adjuvants were highly beneficial in the skin cancer cells when compared with MAL administration alone. AP2‐18 was also at least as effective as CP94 + ALA/MAL co‐administration throughout and significantly better than CP94 supplementation at increasing PpIX fluorescence in MRC5 cells as well as at lower doses where PpIX accumulation was observed to be more limited.ConclusionsPpIX fluorescence levels, as well as PDT cell kill effects on irradiation can be significantly increased by pyridinone iron chelation, either via the addition of CP94 to the administration of a PpIX precursor or alternatively via the newly synthesized combined PpIX prodrug and siderophore, AP2‐18. The effect of the latter compound appears to be at least equivalent to, if not better than, the separate administration of its constituent parts, particularly when employing MAL to destroy skin cancer cells. AP2‐18 therefore warrants further detailed analysis, as it may have the potential to improve dermatological PDT outcomes in applications currently requiring enhancement. Lasers Surg. Med. 50:552–565, 2018. © 2018 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

In vitro cytotoxicity of covalently protected layered molybdenum disulfide

Layered molybdenum disulfide (MoS2) is in forefront of materials research. MoS2 has been reported to display various unique electronic, catalytic and sensing properties. Functionalization of MoS2 is common as it can improve biocompatibility, electrochemical behaviour, catalytic performance, and sensing abilities. In this study, a cytotoxicity study of MoS2 functionalized with an organic molecule, thiobarbituric acid (TBA) was conducted. The functionalization of MoS2 with TBA (MoS2-TBA) is unique as it was reported to retain metallic 1T-type character which is important for electrochemical applications. In addition, MoS2-TBA could possibly be utilized to a wider range of applications as it can be further modified due to the presence of alcohol groups. Thus, investigating the toxicity of MoS2-TBA is essential since it has promising properties for several applications. Functionalized MoS2-TBA is expected to have different toxicity profile than non-functionalized MoS2 as several physicochemical properties may have changed after functionalization. The cytotoxicity study has been conducted via two common cell viability assays which are methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and water-soluble tetrazolium salt (WST-8) assay. Human lung carcinoma epithelial cell line (A549) was chosen for this study it is highly probable for nanomaterials to enter the lungs and interact with the body via inhalation through the respiratory tract from the environment. This study has found that MoS2-TBA has become less toxic as compared to MoS2.

Vivo-morpholino oligomers strongly inhibit dengue virus replication and production.

Dengue virus (DENV) infection is a worldwide public health problem, which can cause severe dengue hemorrhagic fever (DHF) and life-threatening dengue shock syndrome (DSS). There are currently no anti-DENV drugs available, and there has been an intensive search for effective anti-DENV agents that can inhibit all four DENV serotypes. In this study, we tested whether vivo-morpholino oligomers (vivo-MOs), whose effect on DENV infection has not previously been studied, can inhibit DENV infection. Vivo-MOs were designed to target the top of 3' stem-loop (3' SL) in the 3' UTR of the DENV genome and tested for inhibition of DENV infection in monkey kidney epithelial (Vero) cells and human lung epithelial carcinoma (A549) cells. The results showed that vivo-MOs could bind to a DENV RNA sequence and markedly reduce DENV-RNA, protein, and virus production in infected Vero and A549 cells. Vivo-MOs at a concentration of 4 µM could inhibit DENV production by more than 104-fold when compared to that of an untreated control. In addition, vivo-MOs also inhibited DENV production in U937 cells and primary human monocytes. Therefore, vivo-MOs targeting to the 3' SL in the 3' UTR of DENV genomes are effective and have the potential to be developed as anti-DENV agents.

An in vitro cytotoxicity assessment of graphene nanosheets on alveolar cells

The collection of intrinsic properties possessed by graphene family nanomaterials (GFNs) results in their continuous exploitation for biomedical applications. The materials biomedical potential has motivated an upsurge in green preparation routes for the production of graphene like materials with limited toxicity. A number of bio-friendly reducing agents have been utilized for the preparation of chemically reduced graphene oxide (GO), and their resulting cytotoxic effects examined. However, the toxicology effects of one of the first biomolecules implemented for the reduction of GO, ascorbic acid (AA) has yet to be investigated. Herein, the toxicity of three distinct GFNs; GO, hydrazine reduced GO (H.rGO) and AA.rGO, prepared through diverse chemical routes are studied, to demonstrate the cytotoxic activity of a green reducer, in comparison to an established reduction method using hydrazine hydrate. The variation in atomic structure of GO, H.rGO and AA.rGO resulting from different synthesis techniques demonstrates the dependence of toxicity on particle shape and size. All GFNs induced high levels of alveolar cell toxicity. Interaction of AA.rGO with the A549 human lung epithelial carcinoma cell line resulted in increased leakage of lactate dehydrogenase, indicative of diminished cell membrane integrity. The uncharacteristic shape of the AA.rGO may be responsible for this proliferated release of the essential protein. The presented data therefore demonstrates that modification of synthetic processes significantly alter the biological activities of GFNs.

Cytotoxicity of Group 5 Transition Metal Ditellurides (MTe2 ; M=V, Nb, Ta).

Much research effort has been put in to study layered compounds with transition metal dichalcogenides (TMDs) being one of the most studied compounds. Due to their extraordinary properties such as excellent electrochemical properties, tuneable band gaps, and low shear resistance due to weak van der Waals interactions between layers, TMDs have been found to have wide applications such as electrocatalysts for hydrogen evolution reactions, supercapacitors, biosensors, field-effect transistors (FETs), photovoltaics, and lubricant additives. In very recent years, Group 5 transition metal ditellurides have received an immense amount of research attention. However to date, little has been known of the potential toxicities posed by these materials. As such, we conducted the cytotoxicity study by incubating various concentrations of the Group 5 transition metal ditellurides (MTe2 ; M=V, Nb, Ta) with human lung carcinoma epithelial A549 cells for 24 hours and the remaining cell viabilities after treatment was measured. Our findings indicate that VTe2 is highly toxic whereas NbTe2 and TaTe2 are deemed to exhibit mild toxicities. This study constitutes an exemplary first step towards the understanding of the Group 5 transition metal ditellurides' toxicity effects in preparation for their possible future commercialisation.

Abstract 3665: Tumor Treating Fields (TTFields) plus anti-PD-1 therapy induce immunogenic cell death resulting in enhanced antitumor efficacy

Abstract Tumor Treating Fields (TTFields) are an effective anti-neoplastic treatment modality delivered via noninvasive application of low intensity, intermediate frequency, alternating electric fields. TTFields is approved for the treatment of both newly diagnosed and recurrent glioblastoma. TTFields interrupt cancer cell mitosis by disrupting microtubules and septin filaments, which play key roles in mitosis. The mitotic effects of TTFields include abnormal chromosome segregation and ER stress, which trigger different forms of cell death. We evaluated the in vitro and in vivo effects of TTFields combined with an immune checkpoint inhibitor (anti-PD1) on immunogenic cell death. Murine Lewis lung carcinoma (LLC) and ovarian surface epithelial (MOSE) cells were treated with TTFields using the inovitroTM system. Levels of calreticulin (CRT) on the surface of treated cells and intracellular ATP levels were evaluated using flow cytometry. High mobility group box 1 (HMGB1) secretion was measured using an ELISA assay. Mice were implanted with LLC cells were treated with TTFields, anti-PD-1, or a combination of the two modalities. Tumor volume was monitored; flow cytometry analysis was performed for phenotypic characterization of infiltrating immune cells. TTFields induced elevated cell surface expression of CRT, decreased intracellular ATP levels, and promoted HMGB1 secretion. In vivo, the combined treatment of lung tumor-bearing mice with TTFields plus anti-PD-1 led to a significant decrease in tumor volume compared to anti-PD-1 alone or to the control group. Significant increases in CD45+ tumor infiltrating cells were observed in the TTFields plus anti-PD-1 group. Infiltrating cells demonstrated a significant upregulation of surface PD-L1 expression. Both F4/80+CD11b+ cells and CS11c+ cells exhibited higher tumor infiltration and elevated PD-L1 expression as compared to infiltrating immune cell in the control group. Our results demonstrate that TTFields treatment potentiates immunogenic cell death in cancer cells. Combining TTFields with specific immunotherapies such as anti-PD-1 may enhance antitumor immunity and result in increased tumor control. Citation Format: Tali Voloshin, Orna Tal-Yitzhaki, Noa Kaynan, Moshe Giladi, Anna Shteingauz, Mijal Munster, Roni Blat, Yaara Porat, Rosa S. Schneiderman, Shay Cahal, Aviran Itzhaki, Eilon D. Kirson, Uri Weinberg, Yoram Palti. Tumor Treating Fields (TTFields) plus anti-PD-1 therapy induce immunogenic cell death resulting in enhanced antitumor efficacy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3665. doi:10.1158/1538-7445.AM2017-3665

Abstract 5524: Gene 33/Mig6 regulates apoptosis and the DNA damage response through independent mechanisms

Abstract Gene 33 (Mig6, ERRFI1) is an inducible adaptor/scaffold protein whose expression can be induced by both stress and mitogenic signals. It contains multiple domains for protein-protein interaction and is involved in a broad spectrum of cellular functions including cell proliferation, cell migration, cell apoptosis, and cell senescence. Many functions of Gene 33 are attributable to its ability to bind to the kinase domain of the EGFR family receptor tyrosine kinases thereby inhibiting their activities. Gene 33 also binds and activates tyrosine kinase c-Abl and is involved in other signing pathways such as NF-κB, HGF, and JNK. Growing evidence at the genetic, cellular, and animal levels indicates that Gene 33 functions as a tumor suppressor in the lung. Gene 33 has been shown to promote apoptosis. A recent study has linked this protein to the DNA damage response induced by hexavalent chromium [Cr(VI)]. In the current study we find that ectopic expression of Gene 33 strongly induces apoptosis by both activating the c-Abl/p73 pathway and inhibiting the EGFR/AKT pathways in BEAS-2B lung epithelial cells and A549 lung carcinoma cells. Surprisingly, ectopic expression of Gene 33 also triggers the DNA damage response in an ATM-dependent fashion and through pathways with and without association with apoptosis. We observed striking presence of Gene 33 in the nucleus and chromatin, which is in contrast with the belief that Gene 33 is an exclusively cytoplasmic protein. Our data show that chromatin localization of Gene 33 is partially dependent on the EBD motif of Gene 33, a domain required for interaction with EGFR and c-Abl. Our data also indicate that Gene 33 may regulate chromatin targeting of c-Abl and EGFR. Furthermore we find that both endogenous and ectopically expressed Gene 33 strongly interact with histone H2AX. Our data have revealed potential nuclear/chromatin-associated mechanisms that underline the function of Gene 33 in apoptosis and the DNA damage response. Citation Format: Cen Li, Soyoung Park, Xiaowen Zhang, Leonard Eisenberg, Hong Zhao, Zbigniew Darzynkiewicz, Dazhong Xu. Gene 33/Mig6 regulates apoptosis and the DNA damage response through independent mechanisms [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5524. doi:10.1158/1538-7445.AM2017-5524

Abstract 1123: Sphingolipids transfer proteins (GLTP and CPTP) regulate the neoplastic progression of colon and breast cancer cells

Abstract Sphingolipids have previously been shown to affect cancer progression by regulating cell death (apoptosis) and survival (autophagy). While apoptosis is promoted by elevations in sphingosine and ceramide, increases in sphingosine-1-phosphate and ceramide-1-phosphate can tip the balance toward survival. Changes in the expression of sphingolipid transfer proteins such as GLTP (glycolipid transfer protein) and CPTP (ceramide-1-phosphate transfer protein) have been shown to modulate cell shape change and regulate pro-inflammatory cytokine release, respectively, in human cancer epithelial cell lines. To gain a better understanding of the situation, we investigated further. We discovered that GLTP overexpression inhibits the growth of human colon carcinoma cell lines (HT-29; HCT-116), but spares normal colonic epithelial cells (HCEC) largely due to growth arrest at the G0/G1 cell cycle checkpoint. Mechanistically, we found that GLTP overexpression modulated cell cycle progression by upregulating Kip1/p27 and Cip1/p21 protein and mRNA levels, while decreasing CDK2, CDK4, cyclin E and cyclin D1 protein levels. The cell cycle arrest and growth inhibition induced by GLTP overexpression led to apoptotic cell death of HT-29 cells. Neither GLTP nor CPTP overexpression induced apoptosis in breast cancer cell lines (T47D and HTB-126). However, depletion of CPTP, but not GLTP, induced autophagy as indicated by increased LC3 transcript and autophagosome levels (GFP-LC3-II puncta). In the breast cancer cell lines, increased transcript levels of cPLA2 and CERK were detected consistent with the earlier pro-inflammatory effect reported for CPTP depletion in A549 lung epithelial carcinoma cells. Our findings shed light on the mechanistic ways by which GLTP and CPTP affect apoptosis and autophagy to impact cancer progression. Future in vivo studies are planned to ascertain the role(s) of these new players in cancer development and therapy. [Support: NIH GM45928, HL-125353, Paint-the-Town-Pink (PTTP) Cancer Funds, Hormel Fdn] Citation Format: Shrawan K. Mishra, Rhoderick E. Brown. Sphingolipids transfer proteins (GLTP and CPTP) regulate the neoplastic progression of colon and breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1123. doi:10.1158/1538-7445.AM2017-1123

Rapid microwave assisted synthesis and antiproliferative evaluation of novel steroidal thiazole derivatives

Three series of novel steroidal thiazole derivatives were synthesized by microwave assisted one-pot reaction from pregnenolone, testosterone and estrone, respectively. Their structures were characterized by IR, NMR and HRMS, and the antiproliferative activities of all the synthesized compounds against human cervical carcinoma (HeLa), human liver carcinoma(HepG2), human lung carcinoma(A549), nasopharyngeal carcinoma(CNE2) and normal kidney epithelial cells(HEK293T) were screened. Among the compounds, 1-estron-17′-ylidene-2-[4′-(p-bromophenyl)-2′-thiazol] hydrazone(12) displayed distinct antiproliferative activity against all the tested cancer cell lines and was almost inactive to normal kidney epithelial cells.

In vitro detection of diesel exhaust particles induced human lung carcinoma epithelial cells damage and the effect of resveratrol.

People are taking up antioxidants in their daily diet and being exposed to a potential diesel exhaust particles (DEP)-containing environment. Thus it is important to study in vitro cellular responses when cells are exposed to DEP with or without antioxidant treatment. The investigation of DEP and resveratrol (RES) on cellular biophysical and biochemical changes is needed to better understand the mechanisms of DEP and RES in mammalian cells. A combination of two non-invasive techniques (atomic force microscopy, AFM, and Raman spectroscopy, RM) and multimodal tools were applied to evaluate the biophysical, biochemical alterations and cytokine, membrane potential and cell cycle of cells with or without RES pretreatment to different times of DEP exposure. AFM results indicated that RES protected cells from DEP-induced damage to cytoskeleton and cell architectures, and noted that RES treatments also attenuated DEP-induced alterations in cell elasticity and surface adhesion force over DEP incubation time. RM monitored the changes in characteristic Raman peak intensities of DNA and protein over the DEP exposure time for both RES and non-RES treated groups. The cytokine and chemokine changes quantified by Multiplex ELISA revealed that the inflammatory responses were enhanced with the increase in DEP exposure time and that RES enhanced the expression levels of cytokine and chemokine. This work demonstrated that significant biophysical and biochemical changes in cells might be relevant to early pathological changes induced by DEP damage. Copyright © 2016 John Wiley & Sons, Ltd.

Induction of cancer-associated fibroblast-like cells by carbon nanotubes dictates its tumorigenicity.

Tumor microenvironment has been recognized as a key determinant of tumor formation and metastasis, but how tumor microenvironment is affected by nanomaterials is essentially unknown. Here, we investigated whether carbon nanotubes (CNTs), a widely used nanomaterial with known carcinogenic potential, can affect cancer-associated fibroblasts (CAFs), which are a key component of tumor microenvironment that provides necessary support for tumor growth. We show for the first time that single-walled CNT and to a lesser extent multi-walled and its COOH-functionalized form induced CAF-like cells, which are non-tumorigenic in animals, but promote tumor growth of human lung carcinoma and CNT-transformed lung epithelial cells. The mechanism by which CNT-induced CAF-like cells promote tumor growth involved the acquisition of cancer stem cells (CSCs) in cancer population. Gene knockdown experiments showed that an expression of podoplanin on CAF-like cells is essential for their effects, indicating the functional role of CAF-like cells and podoplanin in CNT tumorigenic process. Our findings unveil a novel mechanism of CNT-induced carcinogenesis through the induction of CAF-like cells that support CSCs and drive tumor formation. Our results also suggest the potential utility of podoplanin as a mechanism-based biomarker for rapid screening of carcinogenicity of CNTs and related nanomaterials for their safer design.

Integrins are not essential for entry of coxsackievirus A9 into SW480 human colon adenocarcinoma cells.

BackgroundCoxsackievirus A9 (CV-A9) is a pathogenic enterovirus type within the family Picornaviridae. CV-A9 infects A549 human epithelial lung carcinoma cells by attaching to the αVβ6 integrin receptor through a highly conserved Arg-Gly-Asp (RGD) motif, which is located at the exposed carboxy-terminus of the capsid protein VP1 detected in all studied clinical isolates. However, genetically-modified CV-A9 that lacks the RGD motif (CV-A9-RGDdel) has been shown to be infectious in some cell lines but not in A549, suggesting that RGD-mediated integrin binding is not always essential for efficient entry of CV-A9.MethodsTwo cell lines, A549 and SW480, were used in the study. SW480 was the study object for the integrin-independent entry and A549 was used as the control for integrin-dependent entry. Receptor levels were quantitated by cell sorting and quantitative PCR. Antibody blocking assay and siRNA silencing of receptor-encoding genes were used to block virus infection. Peptide phage display library was used to identify peptide binders to CV-A9. Immunofluorescence and confocal microscopy were used to visualize the virus infection in the cells.ResultsWe investigated the receptor use and early stages of CV-A9 internalization to SW480 human epithelial colon adenocarcinoma cells. Contrary to A549 infection, we showed that both CV-A9 and CV-A9-RGDdel internalized into SW480 cells and that function-blocking anti-αV integrin antibodies had no effect on the binding and entry of CV-A9. Whereas siRNA silencing of β6 integrin subunit had no influence on virus infection in SW480, silencing of β2-microglobulin (β2M) inhibited the virus infection in both cell lines. By using a peptide phage display screening, the virus-binding peptide identical to the N-terminal sequence of HSPA5 protein was identified and shown to block the virus infection in both A549 and SW480 cell lines. HSPA5 was also found to co-localize with CV-A9 at the SW480 cell periphery during the early stages of infection by confocal microscopy.ConclusionsThe data suggest that while αVβ6 integrin is essential for CV-A9 in A549 cell line, it is not required in SW480 cell line in which β2M and HSPA5 alone are sufficient for CV-A9 infection. This suggests that the choice of CV-A9 receptor(s) is dependent on the tissue/cellular environment.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-016-0619-y) contains supplementary material, which is available to authorized users.

The antitumor activity of alternating electric fields (TTFields) in combination with immune checkpoint inhibitors.

e14570 Background: Tumor Treating Fields (TTFields) are an effective anti-neoplastic treatment modality delivered via application of low intensity, intermediate frequency, alternating electric fields. This therapy is approved for the treatment of patients with glioblastoma. Previous investigations have shown that some of the outcomes of mitosis under TTFields application include abnormal chromosome segregation and ER stress which trigger different forms of cell death. In this study we evaluated whether TTFields induced cancer cell death can be perceived as immunogenic by the immune system, and explored the possibility of combining TTFields with immune-modulating drugs. Methods: Murine Lewis lung carcinoma (LLC) and ovarian surface epithelial (MOSE) cells were treated with TTFields using the inovitro system. The exposure of calreticulin (CRT) on the surface of treated cells was evaluated using flow cytometry. HMGB1 secretion was measured using ELISA assay. For in-vivo experiments, mice were implanted with LLC cells, and treated with TTFields, anti-PD-1 or combination of the two modalities. Changes in tumor volume were monitored and flow cytometry analysis was performed for phenotypic characterization of tumor infiltrating immune cells. Results: We demonstrate that application of TTFields leads to surface exposure of CRT and also promotes release of HMGB1. In vivo, the combined treatment of TTFields and anti-PD-1 led to a significant decrease in tumor volume compared to control group and to animals treated with anti-PD-1 alone. Significant increase in CD45+ tumor infiltrating cells was observed in the TTFields+anti-PD-1 combination treatment group. This cell population demonstrated a significant upregulation of PD-L1 expression on the cell surface. Specifically, this upregulation was observed in both F4/80+CD11b+ cells and CD11c+ cells whereas no effect on the infiltration pattern of these immune cell populations was noted. Conclusions: Our results demonstrate that TTFields application potentiates immunogenic cell death in cancer cells and that combining TTFields with specific immunotherapies such as anti-PD-1 might achieve tumor control by further enhancing antitumor immunity.

Cytotoxicity of Graphene Derivatives and Exfoliated Transition-Metal Dichalcogenides (MoS2, WS2, and WSe2)

Many 2D nanomaterials have been demonstrated to possess amazing physiochemical properties and are likely to be incorporated in future consumer products. Therefore, there is a urgent need to ensure that these nanomaterials do not cause adverse health effects. In this study, the cytotoxicity of three common exfoliated TMDs (exTMDs), namely MoS2, WS2, and WSe2, were compared with that of graphene oxides and halogenated graphenes to determine which group of 2D nanomaterials would display better biocompatability. Cell viability assessments using methylthiazolyldiphenyl-tetrazolium bromide (MTT) and water-soluble tetrazolium salt (WST-8) assays on human lung carcinoma epithelial cells (A549) exposed to varying concentrations of these 2D nanomaterials revealed that MoS2 and WS2 nanosheets induced very low cytotoxicity to A549 cells, even at high concentrations. On the other hand, WSe2 was found to exhibit dose-dependent toxicological effects on A549 cells, reducing cell viability to 31.8 % at the maximum concentration of 400 μg mL−1. In comparison with graphene oxides and halogenated graphenes, MoS2 and WS2 were much less hazardous, whereas WSe2 showed similar degree of cytotoxicity, suggesting that MoS2 and WS2 might be suitable and safe materials for potential applications. However, future in-depth studies should be built upon this first work on the in vitro cytotoxicity of MoS2 and WS2 to ensure that they do not pose acute toxicity. Lastly, nanomaterial-induced interference control experiments indicated that MTT assay is not suitable for measuring the cytotoxicity of exTMDs as inflated absorbance values will be obtained due to undesired reaction between exTMDs and the MTT assay viability markers, giving false impressions that the materials are less toxic.

Synthesis and Evaluation of Some New B-Norcholesteryl Thiazole Compounds as Anticancer Agents

Some B-norcholesteryl thiazole compounds were synthesized and their structures were determined by IR, NMR and HRMS. The antiproliferative activity of the compounds against human lung carcinoma (A549), cervical carcinoma (HeLa), liver carcinoma (HEPG2) and normal kidney epithelial (HEK293T) cells was assayed. The results showed that some compounds with a structure of N-methylthiazole showed distinct antiproliferative activity against A549 and HEPG2 cells. The compounds with a structure of N-phenylthiazole displayed a selective antiproliferative activity against HeLa cells and were almost inactive to HEK293T cells. The research provided a theoretical reference for the exploration of new anti-cancer agents.

A novel surgical mesh suitable for laparoscopy, studied on animal model

Reconstruction of the abdominal wall with mesh is a widely used surgical procedure. The non-absorbable meshes tend to cause numerous side-effects. The aim of the author was to produce an absorbable, polymer-based mesh that possesses appropriate chemical, mechanical and biological properties. A three-dimensional, biocompatible mesh was produced from poly-vinyl-alcohol using reactive electrospinning. Toxicity and cell-mesh interactions were tested using human lung carcinoma epithelial cells (A-549), and in vivo tests were conducted in 42 male Wistar rats at the 1-5, 7 and 14 postoperative days (3 rats/groups). In the in vitro tests poly-vinyl-alcohol was biocompatible. In the in vivo tests no major complication was associated with the mesh made of poly-vinyl-alcohol. The author concludes that this polymer mesh is biocompatible, it does not damage the surrounding tissues and integrates well with them.