Epithelial Lung Cancer Cell Lines Research Articles

Anticancer activity of peptide W-0803 derived from Anoplophoa glabripennis

Natural compounds are known as a resource of anti-cancer agents. Anti-cancer capacity toward human epithelial lung cancer cell lines (A549, H460) was examined with and without treatment of trypsin on the extract of Anoplophora glabripennis. IC50 values without trypsin treatment were about 21.3 and 25.0 μg/mL for H460 and A549, respectively. When the extract was treated with trypsin, the IC50 values were 16.0 and 15.6 μg/mL for H460 and A549, respectively, indicating that the trypsin treatment increased the anti-cancer capacity. Because trypsin treatment increased the capacity, the extract was treated with trypsin to isolate the peptide W-0803 which has lysin (K) in C-terminal and α-helix structures. With treatment of W-0803, the cell viability decreased dose-dependently for H460 and A549 cells. Apoptosis analysis showed that the cell death with the treatment of peptide W-0803 was mainly by apoptosis. The wound-healing assay also showed that the peptide W-0803 has an inhibitory capability on cell migration of H460 and A549 cells. All these results suggest that the peptide W-0803 is an anti-cancer agent for lung cancer cell treatment.

Exploring the synthesis, structure and bioactivity of pyrimidine carboxylic acid-derived organic antimony (V) complexes: Cytostatic and antimicrobial evaluations

Eight new organoantimony complexes, R3SbL2 (R = Ph, p-MePh; L = L1, L2) and (R3Sb)2OL2 (R = pHPh, m-MePh; L = L1, L2), [Ph3Sb(C5H3N2O2)2] (1), [(Ph3Sb)2O(C5H3N2O2)2] (2), [(p-tol)3Sb(C5H3N2O2)2] (3), [(m-tol)6Sb2O(C5H3N2O2)2] (4), [Ph3Sb(C5H3N2O2)2] (5), [(Ph3Sb)2O(C5H3N2O2)2] (6), [(p-tol)3Sb(C5H3N2O2)2] (7) and [(m-tol)6Sb2O(C5H3N2O2)2] (8), were designed and synthesized by the reaction of 4-pyrimidine carboxylic acid (HL1) or 5-pyrimidine carboxylic acid (HL2) with organoantimony precursors. All the complexes were characterized by elemental analysis, FT-IR, NMR (1H and 13C), and single crystal X-ray diffraction analysis. Crystal structure results reveal that the central antimony atom of all the complexes displays a triangular biconical geometry. Among them, complexes 1, 3, 5 and 7 show a monomeric structure, while complexes 2, 4, 6 and 8 have a dimeric structure with oxygen bridges. The in vitro cytostatic activities of the eight complexes against a human alveolar basal epithelial lung cancer cell lines (A549) and hepatocellular carcinoma cell lines (HepG-2) were investigated, and the results showed that the organoantimony derivatives with the tolyl groups (3, 4, 7, 8) exhibited higher in vitro cytostatic activities compared with those with the phenyl groups (1, 2, 5, 6). At the same time, bacteriostatic tests were conducted to assess the inhibitory effects of complexes 1–8 against three prevalent plant pathogenic fungi and one bacterium. The results revealed that complex 4 exhibited significant fungicidal activities, on the other hand, complex 3 exhibited higher antibacterial activity against MRSA. These results will provide significant guidance for the further exploration of potential efficient biological activity complexes.

Antiproliferative and immunomodulative potential of Citrullus colocynthis and its bioactive compounds in human lymphocytes and lung cells

Ethnopharmacological relevanceCitrullus colocynthis (L.) Schrad is a member of the Cucurbitaceae plant family which has been used in traditional medicine for the treatment of lung diseases such as asthma and bronchitis. Aim of the studyThe study was conducted to investigate antiproliferative and immunomodulating effects of C. colocynthis and isolated cucurbitacins on human T lymphocytes and lung epithelial cells in order to evaluate their potential in the treatment of airway diseases. Materials and methodsDifferent concentrations of an ethanolic extract of C. colocynthis fruits and cucurbitacins B (CuB), E (CuE) and E-glucopyranoside (CuE-Glu) were analysed for their cytotoxicity and immunomodulatory potential on Peripheral Blood Mononuclear Cells (PBMCs) of healthy donors and on the epithelial lung cancer cell line A549. Viability and proliferation were tested using WST1 and CFSE assays. Flow cytometric analysis of AnnexinV/PI staining was used to investigate cell death through apoptosis/necrosis. Effects on regulatory mechanisms of T lymphocytes, such as CD69 and CD25 marker activation, cytokine production of the cytokines interleukin 2 (IL2), tumor necrosis factor α (TNFα) and interferon γ (IFNy) were also analysed via flow cytometry. Influences on the activator protein 1 (AP1), nuclear factor of activated T-cells (NFAT) or nuclear factor ‘kappa-light-chain-enhancer’ of activated B-cells (NFκB) pathways were analysed in the Jurkat reporter cell line. Cytokine secretion in A549 cells stimulated with virus-like particles was analysed using the bead-based Legendplex™ assay. ResultsNon-toxic concentrations of C. colocynthis and CuE-Glu showed dose-dependent effects on viability and proliferation in both T lymphocytes and A549 cells. The extracts inhibited lymphocyte activation and suppressed T cell effector functions, which was also shown by lower production of cytokines IL2, TNFα and IFNy. A dose dependent inhibition of the pathways NFκB, NFAT and AP1 in Jurkat cells could be observed. In A549 cells, especially CuE and CuE-Glu showed inhibitory effects on cytokine production following a simulated viral infection. Unglycosylated cucurbitacins were more effective in suppressing the immune function in lymphocytes than glycosylated cucurbitacins, however this activity is limited to cytotoxic concentrations. ConclusionIn our study we could confirm the immunmodulating effect of C. colocynthis and cucurbitacins B, E and E-glucopyranoside in vitro by suppression of different pathways of inflammation and T cell proliferation. Activity in a lung cell model using a virus-like stimulation shows promise for further research regarding cucurbitacins in airway diseases.

High NHLRC2 expression is associated with shortened survival in lung adenocarcinoma.

Certain variants of NHL repeat (named after NCL-1, HT2A and LIN-41)-containing protein 2 (NHLRC2) gene have been linked to severe fibrotic interstitial lung disease in children. The aim of the current study was to evaluate the expression of NHLRC2 in lung cell and tissue samples from patients with lung adenocarcinoma (ADC) and squamous cell carcinoma (SCC). The expression of NHLRC2 in lung tissue samples was studied by immunohistochemistry (102 ADC, 111 SCC), mRNA in situ hybridization (4 ADC, 3 SCC), and Western blot analysis (3 ADC, 2 SCC). The immunohistochemical NHLRC2 expression was measured by image analysis software and the percentage of NHLRC2-positive cancer cells was evaluated by semiquantitative analysis. The immunohistochemical results of NHLRC2 were compared with the clinical and histological characteristics of the patients. NHLRC2 protein levels in primary stromal and epithelial lung cancer cell lines were measured by Western blot analysis. NHLRC2 was mainly expressed in cancer cells and inflammatory cells within the tumor. The NHLRC2 expression evaluated by image analysis method was significantly higher in ADC compared with that in SCC (P<0.001). High NHLRC2 expression was associated with reduced disease specific survival (P=0.002), overall survival (P=0.001), and high mitotic activity (P=0.042) in ADC. Additionally, the proportion of NHLRC2-positive cancer cells analyzed by the semiquantitative method was significantly higher in ADC than in SCC (P<0.001). NHLRC2 expression was higher in lung ADC than in SCC and its expression was associated with poor survival in ADC patients. Further studies are required to clarify the pathogenetic role of NHLRC2 in lung cancer.

Stress granule formation as a marker of cellular toxicity in lung organoids

Cells regulate protein synthesis under stressful circumstances by forming cytoplasmic RNA granules, termed stress granules (SGs). SGs are membrane-less organelles that function as a protective mechanism in response to stress. They function through liquid-liquid phase separation, which is a vital process comprising 2 distinct de-mixed liquid phases. The components of SGs, such as G3BP1, can serve as biomarkers of cell toxicity. Respiratory diseases are among the leading causes of death globally. After the humidifier disinfectant disaster in Korea in 2011, social concerns over respiratory disease-related deaths have been raised, and the importance of inhalation toxicity testing has been emphasized. Traditionally, in vivo animal models have been used to assess inhalation toxicity, but these models still have limitations owing to physiological differences between species. To overcome these limitations, human immortalized lung epithelial and lung cancer cell lines have been used to evaluate lung toxicity in vitro. Human stem cell-derived 3-dimensional organoid technology has recently been developed in various research fields, including lung toxicity. This review discusses SG-related proteins as potential biomarkers for lung toxicity assessment, especially in human lung organoids under stress conditions, such as exposure to toxic chemicals.

P182 VALSARTAN LIMITS PULMONARY AND CARDIAC DAMAGE INDUCED BY SARS–COV–2 IN EXPERIMENTAL MODELS BY REDUCING THE EXPRESSION OF ACE2

BackgroundSARS–CoV–2 primarily affects the respiratory system, but cardiac complications also occur very often. The entry of SARS–CoV–2 into host cells is mediated by the interaction between the viral glycoprotein Spike (S) and the host angiotensin–converting enzyme 2 (ACE2) protein. The use of ACE inhibitors (ACEIs) and angiotensin II type 1 receptor blockers (ARBs) might influence both ACE2 expression and viral infection, but our knowledge about these possible interactions is limited. Aim. To evaluate the effects of ACEIs and ARBs during active viraemia.MethodsWe tested the effects of exposure to the ACEI Lisinopril (100nM and 500nM) and the ARB Valsartan (10µM and 50µM) on three different cell types: green monkey–derived epithelial cells (VERO E6) which are used to replicate SARS–CoV–2, cardiomyocytes derived from hiPSC (iPSC–CMs) as heart model and a lung epithelial cancer cell line (16HBE) as pulmonary model. The SARS–CoV–2 wild strain was inoculated on cell lines for 1 hour. Cell viability was measured 72 hours after infection. The supernatants of the infected cells were collected and titrated by the micro–neutralization assay on VERO E6 cells to verify the presence of the virus. Levels of ACE2 mRNA and protein content on cell lysates were quantified after each treatment by RT–qPCR and western blot, respectively.ResultsACEI and ARB at both concentrations do not affect the viability of the 3 cell lines. Vice versa, viral infection significantly decreases the viability of VERO E6 (–60%, p < 0.0001) and iPSC–CMs (–44%, p < 0.001), while 16HBE cells do not show a cytopathic effect after infection. Viral titration shows that SARS–CoV–2 replicated in cell lines and was actively released into supernatants. Valsartan 50µM decreased virus release in the three cell lines and increased the viability of VERO E6 (+69%, p < 0.01) and iPSC–CMs (+20.5%, p < 0.05) after the infection. Valsartan 50µM also decreases both mRNA (–65% in VERO E6, p < 0.001; –50% in iPSC–CMs, p < 0.05; –60.5% in 16HBE, p < 0.01) and protein levels of ACE2 in all 3 cell lines.ConclusionThe data suggest that ACEIs and ARBs do not worsen the SARS–CoV–2 infection and that Valsartan, by reducing the levels of ACE2 expression, might result protective.

Berberine-loaded liquid crystalline nanoparticles inhibit non-small cell lung cancer proliferation and migration in vitro

Non-small cell lung cancer (NSCLC) is reported to have a high incidence rate and is one of the most prevalent types of cancer contributing towards 85% of all incidences of lung cancer. Berberine is an isoquinoline alkaloid which offers a broad range of therapeutical and pharmacological actions against cancer. However, extremely low water solubility and poor oral bioavailability have largely restricted its therapeutic applications. To overcome these limitations, we formulated berberine-loaded liquid crystalline nanoparticles (LCNs) and investigated their in vitro antiproliferative and antimigratory activity in human lung epithelial cancer cell line (A549). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), trypan blue staining, and colony forming assays were used to evaluate the anti-proliferative activity, while scratch wound healing assay and a modified Boyden chamber assay were carried out to determine the anti-migratory activity. We also investigated major proteins associated with lung cancer progression. The developed nanoparticles were found to have an average particle size of 181.3 nm with spherical shape, high entrapment efficiency (75.35%) and have shown sustained release behaviour. The most remarkable findings reported with berberine-loaded LCNs were significant suppression of proliferation, inhibition of colony formation, inhibition of invasion or migration via epithelial mesenchymal transition, and proliferation related proteins associated with cancer progression. Our findings suggest that anti-cancer compounds with the problem of poor solubility and bioavailability can be overcome by formulating them into nanotechnology-based delivery systems for better efficacy. Further in-depth investigations into anti-cancer mechanistic research will expand and strengthen the current findings of berberine-LCNs as a potential NSCLC treatment option.

Abstract 12174: Valsartan Protects Both Cardiac and Lung Cells From SARS-CoV-2 Infection

Background: Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2) disease (COVID-19) mainly affects the respiratory system, but cardiac complications occur very often. SARS-CoV-2 entry in host cells is mediated by the interaction between the viral Spike (S) glycoprotein and the host angiotensin-converting enzyme 2 (ACE2). The use of angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin II type 1 receptor blockers (ARBs) might influence the expression of ACE2 and viral infection, but not much is known about these interactions. Aim: To evaluate the effects of ACEIs and ARBs during active viraemia. Methods: We tested the ACEI Lisinopril (at 100nM and 500nM) and the ARB Valsartan (at 10uM and 50uM) for one week on two cell types: cardiomyocytes derived from hiPSC (hiPSC-CMs) as heart model and a lung epithelial cancer cell line (16HBE) as pulmonary model. The SARS-CoV-2 wild strain was inoculated in the two treated cell types for one hour. Cell viability was measured 72 hours after infection. Supernatants were collected and titrated to verify the presence of infectious virus using a micro-neutralization assay on VERO-E6 cells. Levels of ACE2 mRNA and protein content on cell lysates were quantified after each treatment by RT-qPCR and western blot, respectively. Results: ACEI and ARB at both concentrations affected the viability of neither hiPSC-CMs nor 16HBE cells in the absence of virus. Vice versa, viral infection significantly decreased viability of both hiPSC-CMs (-46%; p&lt;0,01) and 16HBE (-19%; p&lt;0,05). Viral titration revealed that SARS-CoV-2 replicated in both cell lines and was actively released in supernatants. Importantly, pre-treatment with Valsartan 50uM increased the viability of both hiPSC-CMs and 16HBE after infection, while Lisinopril and the lower dose of Valsartan had neutral effect. Of note, Valsartan 50uM treatment decrease ACE2 mRNA level in both hiPSC-CMs (-47%, p&lt;0,01) and 16HBE (-37%, p&lt;0,01). Also ACE2 protein levels were reduced in cell lysates of hiPSC-CMs and 16HBE treated with Valsartan 50uM. Conclusion: These data suggest that ACEIs and ARBs do not worsen the SARS-CoV-2 infection. On the contrary, Valsartan seems to be protective against SARS-CoV-2 infection, possibly by reducing ACE2 expression.

A Study on the Cytotoxicity of plectranthus amboinicus and bacopa monnieri stem Extracts on Lung Cancer Cell Line

Cancer is one of the leading causes of morbidity and mortality. Many treatment modalities have been developed and the need for plant derived therapeutic is growing due to the adverse effects of chemotherapeutic agents. The present study aims to discover the potential of two herbs Plectranthus amboinicus and Bacopa monnieri. Screening for the bioactives in the methanolic extracts of Plectranthus amboinicus and Bacopa monnieri stem showed the presence of alkaloids, flavonoids, terpenoids, phenols, saponins, carbohydrates. A comparative study on the cytotoxicity of methanolic extracts of the stem of these plants on lung epithelial cancer cell line A549 and normal fibroblast cell line L929 revealed that both extracts possess cancer cell specific cytotoxic activity. However, Bacopa monnieri stem extracts were found to be more effective in inducing cytotoxicity than Plectranthus amboinicus.

Evaluation of l‐2,4‐diaminobutyric acid‐based ultrashort cationic lipopeptides as potential antimicrobial and anticancer agents

AbstractUltrashort cationic lipopeptides (USCLs) are amphiphiles that exhibit antimicrobial and anticancer activity. They are composed of an aliphatic chain attached to a positively charged peptide group consisting of a few amino acid residues. Here, we report the activity of a series of l‐2,4‐diaminobutyric acid (Dab)‐rich USCLs with varying N‐acyl chain length and charge against Escherichia coli, Staphylococcus aureus, Candida albicans, colon, breast, and lung epithelial cancer cell lines. USCLs with chain lengths of 14–18 carbons showed significant antimicrobial activity with C16‐conjugated peptides being the most active against E. coli and S. aureus, and C18‐conjugated peptides being the most active against C. albicans. Doubly charged USCLs with aliphatic tails that are 14 carbon atoms and longer showed more potent anticancer activity compared to their triply charged counterparts. C14‐Dab2‐NH2 displayed little toxicity to normal cells, and selectivity to microbial and cancer cells over normal cells. Its structural simplicity, bioactivity, nontoxicity, and selectivity make it a promising therapeutic agent.

MUC1-C influences cell survival in lung adenocarcinoma Calu-3 cells after SARS-CoV-2 infection

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) induces coronavirus disease 2019 (COVID-19) and may increase the risk of adverse outcomes in lung cancer patients. In this study, we investigated the expression and function of mucin 1 (MUC1) after SARS-CoV-2 infection in the lung epithelial cancer cell line Calu-3. MUC1 is a major constituent of the mucus layer in the respiratory tract and contributes to pathogen defense. SARS-CoV-2 infection induced MUC1 C-terminal subunit (MUC1-C) expression in a STAT3 activation-dependent manner. Inhibition of MUC1-C signaling increased apoptosis-related protein levels and reduced proliferation-related protein levels; however, SARS-CoV-2 replication was not affected. Together, these results suggest that increased MUC1-C expression in response to SARS-CoV-2 infection may trigger the growth of lung cancer cells, and COVID-19 may be a risk factor for lung cancer patients.

Analysis of Single Nucleotide Variants (SNVs) Induced by Exposure to PM10 in Lung Epithelial Cells Using Whole Genome Sequencing.

There are many epidemiological studies asserting that fine dust causes lung cancer, but the biological mechanism is not clear. This study was conducted to investigate the effect of PM10 (particulate matter less than 10 μm) on single nucleotide variants through whole genome sequencing in lung epithelial cancer cell lines (HCC-827, NCI-H358) that have been exposed to PM10. The two cell lines were exposed to PM10 for 15 days. We performed experimental and next generation sequencing analyses on experimental group that had been exposed to PM10 as well as an unexposed control group. After exposure to PM10, 3005 single nucleotide variants were newly identified in the NCI-H358 group, and 4402 mutations were identified in the HCC-827 group. We analyzed these single nucleotide variants with the Mutalisk program. We observed kataegis in chromosome 1 in NCI-H358 and chromosome 7 in HCC-827. In mutational signatures analysis, the COSMIC mutational signature 5 was highest in both HCC-827 and NCI-H358 groups, and each cosine similarity was 0.964 in HCC-827 and 0.979 in the NCI-H358 group. The etiology of COSMIC mutational signature 5 is unknown at present. Well-designed studies are needed to determine whether environmental factors, such as PM10, cause COSMIC mutational signature 5.

Biological Activity of Some Pyrimidine Derivatives: Cytotoxicity and Oxidative Stress Potential in Human Lung Cancer Cell line (A549).

Compounds with pyrimidine ring in their structure have many biological activities including antimicrobial, antiviral and anticancer. Recently, studies related to their synthesis and so their applications have increased. In a previous study, a solid-phase microwave method was used to synthesized 25 new hydroxy- and methoxy-substituent 4,6-diarylpyrimidin-2 (1H) -ol and 4,6 diarylpyrimidin-2 (1H) –thiol compounds. In the present study, as a preliminary estimation of the anticancer activity, the cytotoxicity and oxidative stress induction potential of 6 derivatives that show highest antibacterial activities was evaluated in human lung epithelial cancer cell line (A549). Results of the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity test indicate that pyrimidine derivatives caused concentration-dependent cell death that IC50 values were calculated between 16.7 and 41.5 µg/mL. additonally, pyrimidine derivatives induced significant changes in malondialdehyde (MDA) and glutathione (GSH) levels and catalase (CAT) activity; oxidative stress could be the mechanism of action of the tested pyrimidine derivatives in the cancerous cells. The results could be used to design the further in vivo and in vitro detailed studies to appreciate these pyrimidine derivatives anticancer activity, compare this activity with in-use known drugs, elucidate their mechanisms of action and estimate their safety.

Abstract 16446: Human Mesenchymal Stem Cells of Adult and Fetal Origin Do Not Express ACE2 and are Immune to SARS-CoV-2 Spike Pseudotyped Virus

Background: Angiotensin-converting enzyme 2 (ACE2) expression in the lung has a protective role against injury. However, ACE2 is the main host cell receptor for the Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2) entry. Human mesenchymal stem cells (hMSCs) are currently under investigation for the treatment of pulmonary and cardiac complications of the SARS-CoV-2 disease (COVID-19). However, it is unclear if hMSCs express ACE2 and if they possibly can secrete ACE2. Also, their predisposition to be infected by SARS-CoV-2 is unknown. Aim: To assess if hMSCs of fetal and adult origin constitutively express and secrete ACE2 and if they can be infected by SARS-CoV-2. Methods: We studied 8 hMSC lines: 4 of fetal origin isolated from human placentas (hA-MSC) and 4 of adult origin isolated from bone marrow aspirates (hBM-MSCs). The lung epithelial cancer cell line CALU-3, which expresses high levels of ACE2, was used as positive control. ACE2 expression was evaluated in both standard culture conditions and after 48 hrs of hypoxia exposure (1% O 2 ). We quantified the mRNA levels by RT-qPCR and verified protein content by western blot on both cell lysates and serum-free 48 hrs-conditioned media (MSC-CM). ACE2 levels in MSC-CM were quantified by ELISA assay. Finally, to test MSC viral susceptibility, we produced replication-defective, GFP-tagged retroviral particles bearing the SARS-CoV-2 Spike protein; the pantropic VSV glycoprotein (VSV-G) was used as positive control. Results: ACE2 mRNA level in both fetal and adult MSC was 200-fold lower than in CALU-3. ACE2 protein expression was undetectable in MSC lysates and CM, both by western blot and ELISA. Hypoxia slightly increased ACE2 mRNA level, but not protein level, which still was undetectable. Finally, CALU-3 but not MSC were infected by SARS-CoV-2 Spike pseudovirus, whereas both cell types were susceptible to VSV-G pseudovirus infection. Conclusions: Both fetal and adult MSC do not express significant levels of ACE2 both under normoxia and hypoxia, and therefore the putative therapeutic mechanism of hMSCs in COVID-19 cannot go through the release of ACE2. Importantly, MSCs are immune to SARS-CoV-2 infection and therefore their use appears safe.

Design and synthesis of some barbituric and 1,3-dimethylbarbituric acid derivatives: A non-classical scaffold for potential PARP1 inhibitors

Six series based on barbituric acid 5a-e, 10a-d; thiobarbituric acid 6a-e, 11a-d and 1,3-dimethylbarbituric acid 7a-e, 12a-d were prepared and screened for their in vitro PARP1 inhibition. They revealed promising inhibition at nanomolar level especially compounds 5c, 7b, 7d and 7e (IC50 = 30.51, 41.60, 41.53 and 36.33 nM) with higher potency than olaparib (IC50 = 43.59 nM). Moreover, compounds 5b, 5d, 7a, 12a and 12c exhibited good comparable activity (IC50 = 65.93, 58.90, 66.57, 45.40 and 50.62 nM, respectively). Furthermore, the most active compounds 5c, 7b, 7d, 7e, 12a and 12c against PARP1 in vitro were evaluated in the BRCA1 mutated triple negative breast cancer cell line MDA-MB-436 where 5c and 12c showed higher potency compared to olaparib and result in cell cycle arrest at G2/M phase. 5c and 12c showed apoptotic effects in MDA-MB-436 and potentiated the cytotoxicity of temozolomide in A549 human lung epithelial cancer cell line. Compounds 5c and 12c represent interesting starting points towards PARP1 inhibitors.

In Vitro Photodynamic Therapy of Mononuclear and Dinuclear Iridium(III) Bis(terpyridine) Complexes

Three mononuclear or dinuclear bis(terpyridine) (tpy) iridium(III) complexes bearing pyren-1-yl (pyr) group(s) were synthesized. Their photophysical properties in water and in vitro photodynamic therapy (PDT) effects toward the human lung epithelial cancer cell line A549 and the human epidermal skin cancer cell line A431 were investigated to evaluate the effects of dinuclear versus mononuclear complexes and the impact of the oligoether substituent at the ligand. All complexes possessed pyr-tpy ligand-associated charge transfer (1CT)/1π,π* absorption bands at 350-550 nm, with the dinuclear complex Ir3 showing the much enhanced absorptivity of this band. These complexes exhibited dual emission upon excitation at >430 nm in most cases, with the emitting states being ascribed to 1ILCT (intraligand charge transfer) and 3π,π*/3CT states, respectively. All complexes exhibited relatively weak to moderate cytotoxicity in the dark but high photocytotoxicity upon broadband visible light irradiation. Among them, the dinuclear complex Ir3 showed the highest intracellular reactive oxygen species (ROS) generation and PDT efficiency compared to its mononuclear counterpart Ir1. Introducing an oligoether substituent on one of the tpy ligands in Ir2 also improved its intracellular ROS generation and PDT efficacy compared to those induced by Ir1. Ir3 induced both mitochondrial dysfunction and lysosomal damage upon light activation toward both cell lines, whereas Ir1 and Ir2 caused both mitochondrial dysfunction and lysosomal damage in A431 cells but only lysosomal damage in A549 cells. The dominant cell death pathway induced by Ir1-Ir3 PDT is apoptosis.

Characterization of Amplified Malate Dehydrogenase 1 (MDH1) in the Context of Squamous Cell Lung Cancer

Cancer cells often reroute or stimulate their metabolism to support proliferation, which requires a change in the activity or regulation of key metabolic residues. Malate dehydrogenase 1 (MDH1) catalyzes the NADH‐dependent conversion of oxaloacetate to malate, which can help drive glycolysis through the production of NAD+. MDH1 is amplified in many squamous cell lung carcinomas, the second leading cause of cancer related death, and this amplification is associated with a poor prognosis of 50% reduced survival. The precise role of MDH1 in driving squamous cell lung cancer and the mechanisms that regulate this protein are not yet well understood.Here, we develop a tool to examine the effect of MDH1 amplification in an in vitro model. We aim to examine MDH1 amplification in squamous cell lung carcinoma with an isogenic control. It has been previously shown that MDH1 methylation can suppress glutamine metabolism in pancreatic cancers, and that acetylation of MDH1 alters its activity. We will examine how MDH1 amplification affects squamous cell carcinomas by using biochemical kinetics and cellular methods. We have found that MDH1 amplification influences the production of metabolites in the TCA cycle. Through monoclonal single cell isolation, we will generate a squamous epithelial lung cancer cell line stably overexpressing MDH1. We hypothesize that MDH1 has cysteine residues that can be sensitive to oxidative stress which can alter the normal enzymatic activities of MDH1. Through site‐directed mutagenesis, we will mutate a cysteine residue to a serine residue at the 137 position to examine the possible effect of oxidative stress on MDH1. This residue has been shown to be sensitive to oxidation in the mouse form of MDH1. We will also heterologously express and purify MDH1 mutants and perform kinetic assays to examine the impact of amplified MDH1. By understanding the regulatory strategies of MDH1, we hope to identify new pathways that may be targeted therapeutically.Support or Funding InformationThis work was funded by a Research Scholar Grant, RSG‐19‐075‐01‐TBE, from the American Cancer Society (C.D.S.), National Institutes of Health R00 CA187594 (C.D.S.), U54CA132384 (SDSU) &amp; U54CA132379 (UC San Diego), and IMSD 5R25GM058906 (SDSU), as well as the California Metabolic Research Foundation (SDSU).

Carbon Nanotubes Modulate Activity of Cytotoxic Compounds via a Trojan Horse Mechanism.

Carbon nanotubes (CNTs) are an emerging drug delivery system, but their success is thwarted by potential toxicity concerns. In vitro and in vivo studies imply toxic potential of CNTs, but their potential to influence toxicity of coadministered compounds still remains elusive. Therefore, the present study was conducted to determine the effect of multiwalled CNTs (MWCNTs) on the toxicity of cytotoxic compounds in macrophage (RAW 264.7), lung epithelial (A549), and breast cancer (MCF-7) cell lines. The results suggest that hydrophilicity/lipophilicity of the compounds is a critical parameter. The correlation between log P and enhanced cytotoxic activity followed an inverted U-shaped curve and log P close to 1 exhibited the highest increase in cytotoxicity. Further, the increase in cytotoxicity of drug/MWCNT combinations was proportional to the degree of cellular uptake of MWCNTs. A mathematical model was developed and validated with a test set of compounds. These results suggest that MWCNTs act as a "Trojan horse" for increased intracellular delivery of drugs resulting in enhanced cytotoxic activity.

Integration of dry-column flash chromatography with NMR and FTIR metabolomics to reveal cytotoxic metabolites from Amphoricarpos autariatus

Metabolomics generate a profile of small molecules from plant extracts, which could be directly responsible for bioactivity effects. Using dry-column flash chromatography enabled a rapid and inexpensive method for the very efficient separation of plant extract with a high resolution. This separation method coupled to NMR and FTIR-based metabolomics is applied to identify bioactive natural products. OPLS multivariate analysis method, was used for correlation the chemical composition of the plant extracts, Amphoricarpos autariatus, with the results of cytotoxic activity against Human cervical adenocarcinoma cell line (HeLa) and epithelial lung cancer cell line (A549). In this way, the highest contribution to the cytotoxic activity was recorded for the guaianolide sesquiterpene lactones named amphoricarpolides. The compounds indicated as bioactive after metabolomics analysis were tested, and their cytotoxic activity were confirmed.

Spectroscopic, quantum chemical, molecular docking and in vitro anticancer activity studies on 5-Methoxyindole-3-carboxaldehyde

The FT-IR and FT-Raman spectra of 5-Methoxyindole-3-carboxaldehyde (MICA) molecule were recorded. The experimental and theoretical vibrational wavenumbers were assigned and compared. The non-linear optical (NLO) activity of the molecule was confirmed using second harmonic generation test and compared with theoretical result. The natural bond orbital (NBO) analysis was carried out to evaluate the intramolecular stabilization interactions of the molecule, which are responsible for the bio-activity of the molecule. The molecular electrostatic potential surface, frontier molecular orbitals and Fukui function analysis reveal the charge transfer interactions in the molecule. The total density of states (TDOS), partial density of states (PDOS), and overlap population density of states (OPDOS) studies were performed in order to visualize the molecular orbital contributions of the molecule. The molecular docking analysis was carried out to explore the inhibitory nature of the molecule, which indicates that molecule can acts as a novel inhibitor of lung cancer causing VEGF receptor. In addition, the in vitro cytotoxicity of the molecule confirms the anticancer activity against the human pulmonary epithelial lung cancer cell lines (A549). Hence, the present investigations pave the way for the development of lung cancer drugs.