Human Small Cell Lung Carcinoma Research Articles

Nonviral CRISPR/Cas9 mutagenesis for streamlined generation of mouse lung cancer models

Functional analysis in mouse models is necessary to establish the involvement of a set of genetic variations in tumor development. A modeling platform to facilitate and cost-effectively analyze the role of multiple genes in carcinogenesis would be valuable. Here, we present an innovative strategy for lung mutagenesis using CRISPR/Cas9 ribonucleoproteins delivered via cationic polymers. This approach allows the simultaneous inactivation of multiple genes. We validate the effectiveness of this system by targeting a group of tumor suppressor genes, specifically Rb1, Rbl1, Pten, and Trp53, which were chosen for their potential to cause lung tumors, namely small cell lung carcinoma (SCLC). Tumors with histologic and transcriptomic features of human SCLC emerged after intratracheal administration of CRISPR/polymer nanoparticles. These tumors carried loss-of-function mutations in all four tumor suppressor genes at the targeted positions. These findings were reproduced in two different pure genetic backgrounds. We provide a proof of principle for simplified modeling of lung tumorigenesis to facilitate functional testing of potential cancer-related genes.

Novel Mouse Cell Lines and In Vivo Models for Human High-Grade Neuroendocrine Lung Carcinoma, Small Cell Lung Carcinoma (SCLC), and Large Cell Neuroendocrine Carcinoma (LCNEC).

There is a clear need to expand the toolkit of adequate mouse models and cell lines available for preclinical studies of high-grade neuroendocrine lung carcinoma (small cell lung carcinoma (SCLC) and large cell neuroendocrine carcinoma (LCNEC)). SCLC and LCNEC are two highly aggressive tumor types with dismal prognoses and few therapeutic options. Currently, there is an extreme paucity of material, particularly in the case of LCNEC. Given the lack of murine cell lines and transplant models of LCNEC, the need is imperative. In this study, we generated and examined new models of LCNEC and SCLC transplantable cell lines derived from our previously developed primary mouse LCNEC and SCLC tumors. RNA-seq analysis demonstrated that our cell lines and syngeneic tumors maintained the transcriptome program from the original transgenic primary tumor and displayed strong similarities to human SCLC or LCNEC. Importantly, the SCLC transplanted cell lines showed the ability to metastasize and mimic this characteristic of the human condition. In summary, we generated mouse cell line tools that allow further basic and translational research as well as preclinical testing of new treatment strategies for SCLC and LCNEC. These tools retain important features of their human counterparts and address the lack of LCNEC disease models.

Novel Silver Complexes Based on Phosphanes and Ester Derivatives of Bis(pyrazol-1-yl)acetate Ligands Targeting TrxR: New Promising Chemotherapeutic Tools Relevant to SCLC Management

Bis(pyrazol-1-yl)acetic acid (HC(pz)2COOH) and bis(3,5-dimethyl-pyrazol-1-yl)acetic acid (HC(pzMe2)2COOH) were converted into the methyl ester derivatives 1 (LOMe) and 2 (L2OMe), respectively, and were used for the preparation of silver(I) complexes 3-5. The Ag(I) complexes were prepared by the reaction of AgNO3 and 1,3,5-triaza-7-phosphaadamantane (PTA) or triphenylphosphine (PPh3) with LOMe and L2OMe in methanol solution. All Ag(I) complexes showed a significant in vitro antitumor activity, proving to be more effective than the reference drug cisplatin in the in-house human cancer cell line panel containing examples of different solid tumors. Compounds were particularly effective against the highly aggressive and intrinsically resistant human small-cell lung carcinoma (SCLC) cells, either in 2D and 3D cancer cell models. Mechanistic studies revealed their ability to accumulate into cancer cells and to selectively target Thioredoxin (TrxR), thus leading to redox homeostasis unbalance and ultimately inducing cancer cell death through apoptosis.

THE EXPRESSION OF TLR4, IFN-γ, TGF-β AND TNF-αLL LINE OF HUMAN SMALL CELL LUNG CARCINOMA NCI-H69 AND IN CISPLATIN-RESISTANT SUBLINE NCI-H69/CPR

Aim: to investigate the effect of teichoic acid Staphylococcus aureus for expression of pro-inflammatory cytokines and of TLR4 in a human small cell lung carcinoma cell line NCI-H69, and cisplatin resistant subline NCI-H69/CPR. Methods. Incubation of cells with teichoic acid (1 ng/m) conducted for 2 days. Expression level of TLR4, TGF-β, INF-γ, TNF-α was evaluated by the real time PCR on 7500 Real-Time PCR System, using specific primers and fluorochrome SYBR Green. The reverse transcription reaction was performed with High-Capacity cDNA Reverse Transcription Kit carried out under the conditions: 25 °C - 10 min, 37 °C ‒ 120 min and 85 °C ‒ 5 min. Results. In cell line culture NCI-H69 addition of teichoic acid increased expression of TLR4 by 1.3 times, and IFN-γ – by 1,1 times. Expression of TGF-β and TNF-α was decreased 2.5 and 4.9 times respectively. In cell line culture NCI-H69/CPR the addition of teichoic acid inhibited the expression of all studied parameters. Expression TLR4 decreased by 4.2 times, IFN-γ – by 1.4 times. Expression TGF-β and TNF-α was depressed 1.6 and 1.2 times. The presented data indicate that teichoic acid of bacterial origin provided the effect of modulating the inflammatory effect in lung cancer cell culture, sensitive and resistant to cisplatin. Conclusions. Teichoic acid as a ligand of TLR4 modulates the expression of pro-inflammatory and anti-inflammatory cytokines in small cell lung cancer cell culture and suppresses the expression of TLR4 and all investigated cytokines in the cisplatin-resistant cell line NCI-H69.

Control Efficacy of Entomopathogenic Fungus Purpureocillium lilacinum against Chili Thrips (Scirtothrips dorsalis) on Chili Plant.

Simple SummaryChili thrips (Scirtothrips dorsalis) is an important pest of chili crops and a major vector of viral plant pathogens. Due to the widespread outbreak of thrips, chemical insecticides have been heavily used in the last few decades. To reduce the utilization of chemical pesticides, alternative biocontrol agents such as entomopathogenic fungi have been screened against the thrips. Laboratory screening revealed that 2 insect fungi isolates, Purpureocillium lilacinum TBRC 10638 and Beauveria bassiana BCC48145 were the most effective isolates against chili thrips. The fungus, P. lilacinum TBRC 10638 exhibited the highest efficacy against chili thrips in greenhouse and field trials and thus would be developed as a thrips control agent.In a laboratory assay, it was shown that B. bassiana BCC48145, BCC2660, and P. lilacinum TBRC10638 were the three strains that exhibited the highest insecticidal activity against chili thrips, causing 92.5% and 91.86% and 92.3% corrected mortality, respectively. The fungi B. bassiana BCC48145 and P. lilacinum TBRC10638 were selected for greenhouse spraying. Cytotoxicity test of the extracts from both fungi evaluated against 4 animal cell lines: KB; human oral cavity carcinoma, MCF7; human breast adenocarcinoma, NCI-H187; human small cell lung carcinoma and GFP-expressing Vero cells, showed none-cytotoxic to all cell lines. An efficacy validation in the greenhouse showed that P. lilacinum TBRC 10638 was more effective than B. bassiana BCC48145 and could control the thrips up to 80% when using the fungus at 108 spores/mL. The LC50 values of P. lilacinum TBRC 10638 against chili thrips based on total thrips count from two experiments were 1.42 × 108 and 1.12 × 107 spores/mL when the fungal spores were sprayed once a week. The optimal concentration of P. lilacinum TBRC 10638 spores for effective control of chili thrips was determined at 1.41 × 109 spores/mL. The average efficacy of P. lilacinum TBRC 10638 for thrips control from 3 field trials was 30.08%, 14.39%, and 29.92%. This result was not significantly different from that of the chemical insecticide treatment group, which showed efficacy at 19.27%, 14.92%, and 19.97%. Furthermore, there was no difference in productivity among the different treatment groups. Our results demonstrated that P. lilacinum TBRC 10638 is a promising biocontrol agent that could be used as an alternative to chemical insecticide for controlling chili thrips.

The Effect of VPA Treatment on Radiolabeled DOTATATE Uptake: Differences Observed In Vitro and In Vivo

Background: To improve peptide receptor radionuclide therapy (PRRT), we aimed to enhance the expression of somatostatin type-2 receptors (SSTR2) in vitro and in vivo, using valproic acid (VPA). Methods: Human NCI-H69 small-cell lung carcinoma cells were treated with VPA, followed by [111In]In-DOTATATE uptake studies, RT-qPCR and immunohistochemistry analysis. Furthermore, NCI-H69 xenografted mice were treated with VPA or vehicle, followed by [177Lu]Lu-DOTATATE injection. Biodistribution studies were performed, and tissues were collected for further analysis. Results: VPA significantly increased SSTR2 expression in vitro. In animals, a statistically significant increased [177Lu]Lu-DOTATATE tumoral uptake was observed when VPA was administered eight hours before [177Lu]Lu-DOTATATE administration, but increased tumor SSTR2 expression levels were lacking. The animals also presented significantly higher [177Lu]Lu-DOTATATE blood levels, as well as an elevated renal tubular damage score. This suggests that the enhanced tumor uptake was presumably a consequence of the increased radiotracer circulation and the induced kidney damage. Conclusions: VPA increases SSTR2 expression in vitro. In vivo, the observed increase in tumoral [177Lu]Lu-DOTATATE uptake is not caused by SSTR2 upregulation, but rather by other mechanisms, e.g., an increased [177Lu]Lu-DOTATATE circulation time and renal toxicity. However, since both drugs are safely used in humans, the potential of VPA to improve PRRT remains open for investigation.

Retraction: Structural characterization of centipede oligopeptides and capability detection in human small cell lung carcinoma: inducing apoptosis.

Retraction of ‘Structural characterization of centipede oligopeptides and capability detection in human small cell lung carcinoma: inducing apoptosis’ by JingQuan Zhao et al., RSC Adv., 2019, 9, 10927–10936, https://doi.org/10.1039/C8RA09018A.

Three novel furospirostanol glycosides and a steroidal alkaloid glycoside from the bulbs of Fritillaria camtschatcensis (L.) Ker Gawl., and their cytotoxicity

Three novel steroidal glycosides (1–3) and a previously described steroidal alkaloid glycoside (4) have been isolated from the bulbs of Fritillaria camtschatcensis (L.) Ker Gawl. (Liliaceae). The structures of novel compounds 1–3 were characterized based on NMR spectroscopy and chemical transformations. Compounds 1–3 are furospirostanol glycosides bearing a (3S)-3-hydroxy-3-methylglutaryl moiety at C-26 in the aglycone. Compounds 1–4 were evaluated in terms of their cytotoxic activities toward HL-60 human promyelocytic leukemia cells, A549 human lung adenocarcinoma cells, and SBC-3 human lung small cell carcinoma cells. Only 4 showed moderate cytotoxicity against HL-60, A549, and SBC-3 cells with IC50 values of 22.9, 13.3, and 11.9 µM, respectively. Compound 4 was found to cause necrotic-like cell death in HL-60 cells.

Analysis of Molecular Mechanism of YiqiChutan Formula Regulating DLL4-Notch Signaling to Inhibit Angiogenesis in Lung Cancer.

In order to explore the specific mechanism of YiqiChutan formula (YQCTF) in inhibiting the angiogenesis of lung cancer and its relationship with delta-like ligand 4- (DLL4-) Notch signaling, 30 healthy BALB/c-nu/nu rats were selected and divided into three groups: A549 group (implanted with lung adenocarcinoma cell line A549), NCI-H460 group (implanted with human lung large-cell carcinoma cell line NCI-H460), and NCI-H446 group (implanted with human lung small cell carcinoma cell line NCI-H446) for constructing lung cancer transplanted tumor models. After modeling, the group treated with normal saline was taken as control group, 200 mg/kg of YQCTF was adopted for intervention, and the tumor volume and growth inhibition rate were compared with the vascular targeted inhibitor Sorafenib. HE staining, CD31 fluorescent antibody staining, and microelectron microscopy were adopted to observe the neovascular endothelial cells of the transplanted tumor. The expression of VEGF, HIF-1α, DLL4, and Notch-1 in the transplanted tumors in each group was detected by Western blot and RT-PCR at the protein level or mRNA level. Compared with the control group, the YQCTF-treated group had obvious inhibitory effect on lung cancer transplanted tumor and lung cancer angiogenesis. In the YQCTF-treated group, the density of angiogenesis decreased significantly and the vascular lumen structure also decreased, and the expression levels of VEGF, HIF-1α, DLL4, and Notch-1 in the YQCTF-treated group were all lower than those in the control group. YQCTF could inhibit the growth of lung cancer transplanted tumor through antiangiogenesis, and it could also reduce the amount of angiogenesis in lung cancer transplanted tumor. In addition, the generation of lumen structure was also hindered, which was realized through the VEGF signaling pathway and DLL4-Notch signaling pathway.

Effects of Sclareol Against Small Cell Lung Carcinoma and the Related Mechanism: In Vitro and In Vivo Studies.

This study aimed to investigate the anticancer effects and potential mechanisms of sclareol in a human small cell lung carcinoma (SCLC) cell line. Cell viability was determined by the MTT assay. Cell cycle, apoptosis and caspase activity were evaluated by flow cytometry. Cell cycle and DNA damage related protein expression was determined by western blotting. In vivo evaluation of sclareol was carried out in xenografted tumor mice models. Sclareol significantly reduced cell viability, induced G1 phase arrest and subsequently triggered apoptosis in H1688 cells. In addition, this sclareol-induced growth arrest was associated with DNA damage as indicated by phosphorylation of H2AX, activation of ATR and Chk1. Moreover, in vivo evaluation of sclareol showed that it could inhibit tumor weight and volume in a H1688 xenograft model. Sclareol might be a novel and effective therapeutic agent for the treatment of SCLC patients.

Comparative analysis of TTF-1 binding DNA regions in small-cell lung cancer and non-small-cell lung cancer.

Thyroid transcription factor‐1 (TTF‐1, encoded by the NKX2‐1 gene) is highly expressed in small‐cell lung carcinoma (SCLC) and lung adenocarcinoma (LADC), but how its functional roles differ between SCLC and LADC remains to be elucidated. Here, we compared the genome‐wide distributions of TTF‐1 binding regions and the transcriptional programs regulated by TTF‐1 between NCI‐H209 (H209), a human SCLC cell line, and NCI‐H441 (H441), a human LADC cell line, using chromatin immunoprecipitation‐sequencing (ChIP‐seq) and RNA‐sequencing (RNA‐seq). TTF‐1 binding regions in H209 and H441 cells differed by 75.0% and E‐box motifs were highly enriched exclusively in the TTF‐1 binding regions of H209 cells. Transcriptome profiling revealed that TTF‐1 is involved in neuroendocrine differentiation in H209 cells. We report that TTF‐1 and achaete‐scute homolog 1 (ASCL1, also known as ASH1, an E‐box binding basic helix–loop–helix transcription factor, and a lineage‐survival oncogene of SCLC) are coexpressed and bound to adjacent sites on target genes expressed in SCLC, and cooperatively regulate transcription. Furthermore, TTF‐1 regulated expression of the Bcl‐2 gene family and showed antiapoptotic function in SCLC. Our findings suggest that TTF‐1 promotes SCLC growth and contributes to neuroendocrine and antiapoptotic gene expression by partly coordinating with ASCL1.

ROS-mediated cell death induced by mixed ligand copper(II) complexes of l-proline and diimine: effect of co-ligand

A series of mixed-ligand Cu(II) complexes of the type: (i) [Cu(l-pro)(diimine)(H2O)n](ClO4) (n = 0 or 1), where l-pro is l-proline and diimine is 2,2′-bipyridine (bpy; 1), (ii) 4,4′-dimethyl-2,2′-bipyridine (dmbpy; 2), (iii) 1,10-phenanthroline (phen; 3), (iv) 5,6-dimethyl-1,10-phenanthroline (5,6-dmp; 4), (v) 3,4,7,8-tetramethyl-1,10-phenanthroline (3,4,7,8-tmp; 5), (vi) dipyrido-[3,2-f:2′,3′-h]-quinoxaline (dpq; 6), and (vii) dipyrido[3,2-a:2′,3′-c]phenazine (dppz; 7) have been synthesized and characterized systematically. Complexes 2 and 3 have been structurally characterized. DNA- and protein-binding and cleavage studies revealed that 7 possesses stronger DNA- and protein-binding efficiency and also exhibits self-activating DNA-cleavage in the absence of an activating agent. The hydrophobic complexes 4 and 5 induced self-activated protein cleavage and yielded a single-cleaved fragment each. The cytotoxic property of all the seven complexes was examined by incubation with the human small cell lung carcinoma cell line A549. Complexes 4 and 6 exhibited almost similar cytotoxic properties [IC50 = 1.4 µM (4) and 1.3 µM (6)], which was 9.3 and 10 times, respectively, more efficient than cisplatin (IC50 = 13 µM). Complex 4 induced generation of the highest level of intracellular ROS which correlates with its efficient cytotoxic activity and provides scope for further investigation as a potential anticancer agent.

Antitumor effect of kurarinone and underlying mechanism in small cell lung carcinoma cells.

BackgroundKurarinone, a prenylated flavonone isolated from the roots of Sophora flavescens, is known to be cytotoxic against many human cancer cells but not human small cell lung carcinoma (SCLC) yet. Also, the exact molecular mechanism of kurarinone for induction cytotoxicity remains unknown.Material and methodsWe investigated the effects of kurarinone on cell proliferation, apoptosis, and migration in H1688 SCLC cells. Cell viability was determined by the MTT assay. Apoptotic indices such as cell cycle, mitochondrial membrane potential, cytochrome c release, caspase activity, and death receptors were evaluated by flow cytometry. Transwell migration and invasion assays were also included.ResultsOur results indicated that kurarinone significantly decreased H1688 cell viability and induced the accumulation of sub-G1 fractions by activating caspase-3, -9, and PARP cleavage accompanied by the elevated release of cytochrome c and mitochondrial dysfunction in H1688 cells. Additionally, kurarinone promoted Fas and TRAIL receptor-1 and -2 expression via the caspase-8/Bid pathway, suggesting that kurarinone triggered apoptosis via the mitochondria-mediated and receptor-mediated apoptotic pathways. We also observed that kurarinone repressed migration and invasion capabilities of SCLC cells by suppressing the expression of epithelial-mesenchymal transition-related proteins and matrix metalloproteinases.ConclusionOur findings provided evidence that kurarinone can induce apoptosis in SCLC cells via multiple mechanisms and delayed the cell migration and invasion of SCLC cells.

Abstract 3905: Translational efficacy and safety modeling and simulation to support the clinical development of JNJ-64619178, a PRMT5 inhibitor

Abstract Protein arginine methyltransferase 5 (PRMT5) is an epigenetic enzyme with oncogenic properties. JNJ-64619178 (JNJ178) is a potent, selective, structurally unique PRMT5 inhibitor with good preclinical efficacy in inhibiting the growth of hematologic and solid tumor cell lines. Toxicology studies showed that JNJ178 decreased reticulocytes and neutrophils in rats and dogs. The objectives of translational modeling and simulation were to understand the exposure-response relationship of both safety and efficacy and provide guidance to the first-in-human clinical development of JNJ178. Experimental data for the PK/PD (Pharmacokinetics/Pharmacodynamics) modeling included: plasma concentration after single dose of JNJ178 in non-tumor bearing mice, plasma concentration and PD markers of dimethylation of arginine (%SDMA in plasma and %SMD1/3-Me2 in tumor, respectively) after multiple doses (1 to 10 mg/kg) QD (once daily) of JNJ178 in H1048 (human small cell lung carcinoma) xenografts, and tumor volume in four xenograft mouse models (A427, human lung carcinoma; H441, human lung adenocarcinoma; H520, human squamous cell lung carcinoma; and H1048). Plasma PK were first described by a standard two-compartment model and used as a driver of PD and tumor volume (efficacy). Plasma and tumor PD were modeled using an indirect response model. A hybrid tumor growth coupled with transit compartment mediated tumor killing model was used to fit the tumor volume data. To predict the safety profile of JNJ178, lifespan based indirect response model for erythropoiesis and Friberg myelosuppression model were used to simulate hemoglobin and neutrophil kinetics in human. The PK/PD model described the data well and validated the hypothesis that PD is driven by trough concentration. Based on the exposure-response relationship from the four xenograft models, the trough concentration needed to achieve tumor stasis for mouse was determined. In addition, the level of inhibition in tumor and plasma PD marker that was associated with tumor stasis was identified. Together with human PK parameters scaled using allometry, the dose range needed to achieve target therapeutic exposure for a typical human subject was predicted. Simulation results from erythropoiesis and Friberg myelosuppression models informed the optimal doing schedules for certain dose levels that would allow hematological toxicity to be manageable with <40% reduction in hemoglobin and >1.0 x 109/L neutrophil counts at all times. Overall, a translational modeling and simulation approach that considers safety and efficacy has been instrumental in the design of the first-in-human clinical development of PRMT5 inhibitor JNJ178 regarding selection of dose and schedule. Citation Format: Yue Guo, Nahor Haddish-Berhane, Hillary J. Millar, Tinne Verhulst, Tony Greway, Junguo Zhou, Loeckie DeZwart, Dana Gaffney, Joseph Portale, Dirk Brehmer, An Boeckx, Erika Van Heerde, Daniele Ouellet. Translational efficacy and safety modeling and simulation to support the clinical development of JNJ-64619178, a PRMT5 inhibitor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3905.

Abstract 2897: Dynamic transdifferentiation programs in small cell lung carcinoma

Abstract Tumors must also have the ability to rapidly adjust to a mutable environment during the lifespan of the cell. This form of phenotypic plasticity is typified by biological programs regulated by a host of transcription factors in a well-coordinated “switch” that is initiated by both cell autonomous and microenvironmental cues. We sought to study these dynamics in small cell lung carcinoma (SCLC). First, we used k-means clustering to identify discrete transcriptional programs that are represented across SCLC using gene expression values from 53 and 71 SCLC cell lines and primary tumors, respectively. To investigate these programs further, we used our inventory of 67 patient-derived xenografts (PDX) to establish an ex vivo cell culture system that reconstitutes with high fidelity the gene expression values observed in the PDXs and the primary tumors from which these cells were derived. Our data demonstrates that gene expression groupings or clusters across samples mark distinct SCLC transcriptional states, representing intertumoral variation. However, in an intriguing demonstration of intratumoral heterogeneity, the very same groupings define morphologically and phenotypically distinct subpopulations within individual samples. Subpopulations were represented by suspending aggregates of very small cells (neuroendocrine or NEn), pre-suspension aggregates that are nested above mesenchymal-like cells and appear to give rise to the suspending aggregates (proneural or PN), pleomorphic cells growing as a monolayer composed of larger cells with visible cytoplasm and spindle-like membrane extensions (mesenchymal or MS), and semi-adherent cells that are physically positioned over the latter (neuroepithelial or NEp). WES demonstrated high genetic fidelity across all subpopulations, indicating that the significant morphologic and phenotypic differences were non-genetic. RNAseq and gene ontology was applied to differentially expressed genes to inform classifications. Using network analyses, we identified transcription factors (TFs) that function as central nodes in the gene expression networks of each population. Namely, we show that ASCL1 marks a rapidly proliferating neuroendocrine population (NEn), NEUROD1 marks a primitive neural population (PN) and YAP1 marks a slow-growing mesenchymal population (MS). We also developed a promoter-reporter fluorescent protein system that positions distinctly colored fluorescent proteins under the regulatory control of the promoters of the three TFs. We show that a Markov process models the transition probability of states. Taken together, our results show that human SCLC is comprised of distinct tumor subpopulations characterized by significant, non-genetic plasticity. We posit that understanding of the SCLC intratumoral ecosystem can create new opportunities to evolutionarily steer this tumor toward improved therapeutic results. Note: This abstract was not presented at the meeting. Citation Format: Priyanka Gopal, Kevin Rogacki, Craig D. Peacock, Mohamed E. Abazeed. Dynamic transdifferentiation programs in small cell lung carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2897.

Kdm1a promotes SCLC progression by transcriptionally silencing the tumor suppressor Rest

Small cell lung carcinoma (SCLC) is one of the deadliest cancer types, with a 5-year survival rate less than 10%. Kdm1a/Lsd1 has recently been implicated as a potential therapeutic target for SCLC. However, the underlying molecular mechanism by which Kdm1a promotes the oncogenesis of SCLC has not been fully understood. Kdm1a is significantly elevated in most human SCLC specimens, whereas Rest, a tumor suppressor and neuronal repressive transcriptional factor, is typically inactivated. Knock-out of Kdm1a (Kdm1a-KO) in mouse SCLC cell lines resulted in the suppression of cell growth and soft agar colony formation. RNA-Seq analysis of the Kdm1a-KO cells revealed significant repression of a program of neuroendocrine signature genes, and conversely, a significant upregulation of a network of genes capable of inhibiting tumor cell growth. Rest was identified among the top 10 upregulated genes in Kdm1a-KO cells. The treatment of the SCLC cells with Kdm1a demethylase inhibitors resulted in a dramatic up-regulation of Rest similar to the extent of that in Kdm1a-KO cells. Importantly, accompanying the restored expression of the SCLC signature genes, knock-out of Rest in Kdm1a-KO cells rescued the restricted cell growth and soft agar colony formation. Taken together, these novel findings show that Kdm1a is a key transcriptional repressor of Rest, and that suppression of SCLC progression by the targeted inhibition of Kdm1a depends on the reactivation of Rest, suggesting a new strategy for effective SCLC treatment by targeting the Kdm1a/Rest molecular pathway.

Biomolecular Interaction, Anti-Cancer and Anti-Angiogenic Properties of Cobalt(III) Schiff Base Complexes

Two cobalt(III) Schiff base complexes, trans-[Co(salen)(DA)2](ClO4) (1) and trans-[Co(salophen)(DA)2](ClO4) (2) (where salen: N,N’-bis(salicylidene)ethylenediamine, salopen: N,N’-bis(salicylidene)-1,2-phenylenediamine, DA: dodecylamine) were synthesised and characterised using various spectroscopic and analytical techniques. The binding affinity of both the complexes with CT-DNA was explored adopting UV-visible, fluorescence, circular dichroism spectroscopy and cyclic voltammetry techniques. The results revealed that both the complexes interacted with DNA via intercalation as well as notable groove binding. Protein (BSA) binding ability of these complexes was investigated by absorption and emission spectroscopy which indicate that these complexes engage in strong hydrophobic interaction with BSA. The mode of interaction between these complexes and CT-DNA/BSA was studied by molecular docking analysis. The in vitro cytotoxic property of the complexes was evaluated in A549 (human small cell lung carcinoma) and VERO (African green monkey kidney cells). The results revealed that the complexes affect viability of the cells. AO and EB staining and cell cycle analysis revealed that the mode of cell death is apoptosis. Both the complexes showed profound inhibition of angiogenesis as revealed in in-vivo chicken chorioallantoic membrane (CAM) assay. Of the two complexes, the complex 2 proved to be much more efficient in affecting the viability of lung cancer cells than complex 1. These results indicate that the cobalt(III) Schiff base complexes in this study can be potentially used for cancer chemotherapy and as inhibitor of angiogenesis, in general, and lung cancer in particular, for which there is need for substantiation at the level of signalling mechanisms and gene expressions.

Retracted Article: Structural characterization of centipede oligopeptides and capability detection in human small cell lung carcinoma: inducing apoptosis.

Lung cancer is the most frequent cause of cancer deaths in the world, and smoking is considered as one of the major causes. Small cell lung carcinoma (SCLC) represents a highly malignant and particularly aggressive form, with properties of widespread metastases and poor prognosis. Herein, twenty-five Scolopendra subspinipes mutilans L. Koch Oligopeptides (SSMOs) were isolated and their structures were identified, and the anti-proliferative activity against lung cancer cell lines was evaluated. Results showed that SSMO-5 induced the production of reactive oxygen species (ROS) markedly in NCI-H446 cells. Furthermore, SSMO-5 decreased the mitochondrial membrane potential (MMP) and enhanced the mitochondria-related apoptosis. These results demonstrate that in NCI-H446 cells, the apoptotic and cytotoxic effects of SSMO-5 are mediated by the intrinsic mitochondria-mediated apoptotic pathway, which in turn causes the activation of caspases and increases Bax expression, while decreases Bcl-2 and Bcl-xL expressions and regulates the interaction of p53/MDM2. In conclusion, a ROS-mediated mitochondrial pathway plays an important role in the process of SSMO-5-induced apoptosis against SCLC.

ТЕЙХОЄВА КИСЛОТА МОДУЛЮЄ ЕКСПРЕСІЮ ТОЛЛ-ПОДІБНИХ РЕЦЕПТОРІВ 4 У ЧУТЛИВИХ ТА РЕЗИСТЕНТНИХ ДО ДІЇ ЦИСПЛАТИНУ ЛІНІЯХ КЛІТИН ДРІБНОКЛІТИННОГО РАКУ ЛЕГЕНІ

The Toll-like receptor family plays crucial role in the innate immune system, recognizing the molecular structures associated with pathogens derived from different microbes. TLRs also recognize the molecular structures associated with damage associated with certain diseases, such as cancer. They can either promote tumorigenesis, or suppress it. Besides, the promotion of the tumor cells growth may be associated with the acquisition of drug resistance. The data on the TLR4 expression level changes during this pathological process are controversial. That is why the purpose of our study was to determine the expression level of TLR4 in cisplatin-sensitive and cisplatin-resistant lung cancer cells. Cells line NCI-H69 (human small cell lung carcinoma) and a drug resistant subline of NCI-H69/CPR were used to determine the expression level of TLR4.Incubation of cells with teichoic acid (1 μg / ml) was performed under standard conditions for two days. The level of TLR4 expression in the cell was determined with RT-PCR at 7500 Real-Time PCR Systems ("Applied Bіosystems", USA) and specific primers and asymmetric cyan cationic fluorochrom SYBRGreen (С32Н37N4S+) with spike of consumption at λmax = 488 nm and fluorescence at λmax = 522 nm. It was shown that TLR4 expression level was almost two times lower in WT H69 cells compared with H69 cells resistant to cisplatin. In contrast, teichoic acid oppositely influenced TLR4 expression level: increased by 1.3-times in WT H69 cells whereas decreased 4-times in H69 cells resistant to cisplatin compared with corresponding control samples without teichoic acid. In cisplatin-resistant lung cancer cells found high expression of TLR4 can be inhibited by addingteichoic acida ligand to TLRs.

Synthesis of Novel α‐Aminophosphonate Derivatives, Biological Evaluation as Potent Antiproliferative Agents and Molecular Docking

AbstractA series of novel fluorine containing α‐aminophosphonate derivatives (4 a–4 q) were synthesized in excellent yield and high purity. All these novel Fluorinated α‐aminophosphonate compounds were screened for antiproliferative and apoptosis activity on human non small cell lung carcinoma cells (A549) and human skin melanoma cells (SK‐MEL‐2). Compounds 4 a, 4 b, 4 c, 4 f, 4 i, 4 j and 4 m were found to be more active antiproliferative agent against A549 and SK‐MEL‐2 cells with IC50 value 0.22 to 1.25 μM. Molecular docking study related to binding affinity and binding mode analysis showed that synthesized compounds had potential to inhibit human Topoisomerase IIa enzyme system. Flow cytometric study showed some of these derivatives also induced cell apoptosis and arrest cell cycle at G1 and at G2/M phase. Overall, this study provides future perspective of lead candidate for the future anticancer drug discovery initiatives.