Induced Lung Cancer Research Articles

Bromelain and Olea europaea (L.) leaf extract mediated alleviation of benzo(a)pyrene induced lung cancer through Nrf2 and NFκB pathway.

Lung cancer is the most aggressive as well as deadly form of cancer and most of the lung cancer cases are involved in direct smoking or passive smoking. Oxidative stress and pulmonary inflammation regulated by some transcription factors like Nrf2, NF-κB etc. play important roles in lung cancer. Various combinations of therapies are currently attributed to lung cancer treatment. A plethora of evidence supports that the consumption of plant-derived foods can prevent chronic diseases like cancer. Leaves of olive (Olea europaea L.) are rich in phenolic compounds which are having antioxidant and anti-inflammatory property. Also, bromelain from pineapple juice and from pineapple stem is a potent anti-inflammatory agent. We took a pragmatic approach to prevent carcinogenesis by supplementing the combination of these two extracts. In this study, we have tried to evaluate the amelioration of various hallmarks associated with benzo(a)pyrene-induced lung carcinogenesis upon the combinatorial treatment of ethanolic olive leaf extract (EOLE) and bromelain. We have studied the role of EOLE in amelioration of BaP-induced oxidative stress in the lung. As several reports of anticancer activity of bromelain are available, we have combined EOLE with bromelain to study their protective role against BaP-mediated lung damage. Changes in DNA integrity, LPO level in lung after EOLE-treated animal were examined. Then, we have evaluated the synergistic role of EOLE and bromelain. We have found that EOLE in combination with bromelain was able to increase the translocation of Nrf2 from cytoplasm to nucleus and decrease the translocation of NF-κB from cytoplasm to nucleus. Combination of treatment also reduced the expression of TNFα, IL-6, and some matrix metalloproteinases in lung tissue. Our findings suggest that EOLE and bromelain can synergistically reduce the BaP-induced lung carcinogenesis associated with inflammation and oxidative stress via regulating the expression of various inflammatory markers and also modulating the activity of pulmonary antioxidant armories.

Radon Adsorption in Charcoal

Radon is pervasive in our environment and the second leading cause of lung cancer induction after smoking. Therefore, the measurement of radon activity concentrations in homes is important. The use of charcoal is an easy and cost-efficient method for this purpose, as radon can bind to charcoal via Van der Waals interaction. Admittedly, there are potential influencing factors during exposure that can distort the results and need to be investigated. Consequently, charcoal was exposed in a radon chamber at different parameters. Afterward, the activity of the radon decay products 214Pb and 214Bi was measured and extrapolated to the initial radon activity in the sample. After an exposure of 1 h, around 94% of the maximum value was attained and used as a limit for the subsequent exposure time. Charcoal was exposed at differing humidity ranging from 5 to 94%, but no influence on radon adsorption could be detected. If the samples were not sealed after exposure, radon desorbed with an effective half-life of around 31 h. There is also a strong dependence of radon uptake on the chemical structure of the recipient material, which is interesting for biological materials or diffusion barriers as this determines accumulation and transport.

Discovery of novel dihydroartemisinin-cinnamic hybrids inducing lung cancer cells apoptosis via inhibition of Akt/Bad signal pathway

A series of dihydroartemisinin-cinnamic acid hybrids were designed, synthesized and evaluated. Most of the tested compounds showed enhanced anti-proliferative activities than artemisinin and dihydroartemisinin, among which 16 g had the superior potency with IC50 values ranging from 5.07 μM to 7.88 μM against four tested cancer cell lines. The cell cycle arrest revealed that 16 g induced A549 cell cycle arrest at G0/G1 phase via regulation of G1-related protein expression (Cdk4). Further mechanism studies reveal that 16 g induced A549 cells apoptosis via inhibiting Akt/Bad pathway. Moreover, 16 g depolarized the mitochondria membrane potentials and induced ROS generation in A549. Additionally, 16 g blocked migration of A549 cells in a concentration-dependent manner. What’s more, 16 g is barely nontoxic to zebrafish embryos. Overall, the cell cycle arrest, inhibition of Akt/Bad signal pathway, ROS generation and migration blocked might explain the potent anti-proliferative activities of these compounds.

Germacrone induces lung cancer cell apoptosis and cell cycle arrest via the Akt/MDM2/p53 signaling pathway.

Germacrone (GM) displays a wide range of antitumor, antioxidant and anti-inflammatory effects; however, to the best of our knowledge, the effects of GM on lung cancer cell apoptosis and cell cycle arrest have not been previously reported. The aim of the present study was to investigate discussed the effects of GM on the apoptosis and cycle arrest of lung cancer cells. Cell viability, proliferation and apoptosis were assessed by performing Cell Counting Kit-8, colony formation and TUNEL assays, respectively. Western blotting was performed to detect the expression levels of apoptosis-, cell cycle- and Akt/MDM2 proto-oncogene (MDM2)/p53 signaling pathway-related proteins. Compared with the control group, 50, 100 and 200 µM GM significantly inhibited lung cancer cell proliferation, but significantly induced cell apoptosis and G1/S cell cycle arrest. GM also significantly altered the expression levels of Akt/MDM2/p53 signaling pathway-related proteins compared with the control group. Administration of Akt activator SC79 significantly reversed GM-mediated antiproliferative, proapoptotic and pro-cell cycle arrest effects in lung cancer cells. Therefore, the results of the present study demonstrated that GM induced lung cancer cell apoptosis and cell cycle arrest via the Akt/MDM2/p53 signaling pathway.

Prophylactic effects of Cynara scolymus L. leaf and flower hydroethanolic extracts against diethylnitrosamine/acetylaminoflourene-induced lung cancer in Wistar rats.

The study examines the prophylactic action of artichoke leaf hydroethanolic extract (ALE) and artichoke flower head hydroethanolic extract (AFE) against diethylnitrosamine (DEN)/acetylaminofluorene (AAF)-induced lung cancer in Wistar rats. To chemically induce lung cancer, DEN was injected intraperitoneally twice a week for a fortnight at a dose of 150 mg/kg body weight (b.w.), followed by oral supplementation of AAF four times a week for 3 weeks at a dose of 20 mg/kg b.w. The DEN/AAF-administered rats were orally supplemented with ALE or AFE at a dose of 100 mg/kg b.w. for 17 weeks starting from the 1st week of DEN injection to the 17th week of the experiment. The lung cancerous injuries resulting from DEN/AAF-administration were significantly improved by the treatment with ALE and AFE as observed in histological examination. In addition, there was a significant reduction in lung lipid peroxidation, with resultant elevation in antioxidant enzymatic activity of glutathione-S-transferase, glutathione peroxidase, glutathione reductase, and superoxide dismutase as well as glutathione content in DEN/AAF-supplemented rats treated with ALE and AFE as compared to DEN/AAF-administered control. The lung tumor suppressor protein (p53) and B-cell lymphoma-2 (Bcl-2) mRNA expression significantly increased in the rats treated with ALE and AFE. In conclusion, the finding showed that ALE and AFE produced anti-cancer prophylactic effects against DEN/AAF-induced lung cancer in rats via suppression of oxidative stress and improved apoptotic signal induction.

Interleukin-17A mediates tobacco smoke-induced lung cancer epithelial-mesenchymal transition through transcriptional regulation of ΔNp63α on miR-19.

Interleukin-17A (IL-17A) is an essential inflammatory cytokine in the progress of carcinogenesis. Tobacco smoke (TS) is a major risk factor of lung cancer that influences epithelial-mesenchymal transition (EMT) process. However, the potential mechanism by which IL-17A mediates the progression of lung cancer in TS-induced EMT remains elusive. In the present study, it was revealed that the IL-17A level was elevated in lung cancer tissues, especially in tumor tissues of cases with experience of smoking, and a higher IL-17A level was correlated with induction of EMT in those specimens. Moreover, the expression of ΔNp63α was increased in IL-17A-stimulated lung cancer cells. ΔNp63α functioned as a key oncogene that bound to the miR-17-92 cluster promoter and transcriptionally increased the expression of miR-19 in lung cancer cells. Overexpression of miR-19 promoted EMT in lung cancer with downregulation of E-cadherin and upregulation of N-cadherin, while its inhibition suppressed EMT. Finally, the upregulated levels of IL-17A, ΔNp63α, and miR-19 along with the alteration of EMT-associated biomarkers were found in lung tissues of TS-exposed mice. Taken together, the abovementioned results suggest that IL-17A increases ΔNp63α expression, transcriptionally elevates miR-19 expression, and promotes TS-induced EMT in lung cancer. These findings may provide a new insight for the identification of therapeutic targets for lung cancer.

Sevoflurane induces lung cancer cell apoptosis via inhibition of the expression of miRNA155 gene

Purpose: To determine the apoptotic effect of sevoflurane on lung cancer cells, and the underlying mechanism of action.Methods: Lung adenocarcinoma A549 cells were cultured for 24 h and divided into control group, 1% sevoflurane group and 3% sevoflurane group. The two levels of sevoflurane were provided through a gas monitor connected to each of the sevoflurane groups. The control group was not treated. Flow cytometry was used to analyze A549 cell apoptosis, while qRT-PCR was used for assay of the levels of miRNA155 in A549 cells. The protein expression of Bcl-2 was determined with immunoblotting. The percentage of apoptosis and levels of miRNA155 and Bcl-2 in the two cell lines were compared.Results: Significant differences in miRNA146a level were seen between the 3 % sevoflurane and control groups at 3 h. There was higher apoptosis in the 3 % sevoflurane group, relative to control, but miRNA155 levels in the 3 % sevoflurane group were generally less than that of the control (p < 0.05). There was lower Bcl-2 content in the 3 % sevoflurane group than in control group (p < 0.05).Conclusion: Sevoflurane exerts strong apoptotic and anti-proliferative effects on lung adenocarcinoma A549 cells via a mechanism which may be related to the downregulation of miRNA155, thereby inhibiting the expression of anti-apoptotic protein Bcl-2. This provides a new direction for research on anti-lung adenocarcinoma drugs.
 Keywords: Sevoflurane, Lung cancer cells, Apoptosis, Inhibition, miRNA155, Expression, Induction

Abstract PO-086: Detection of cancer lesions in histopathological lung images using a sparse PCA network

Abstract Purpose: Lung cancer has been the leading cause of cancer-related deaths worldwide. To address the clinical need for efficacious treatments, genetically engineered mouse models (GEMMs) have become integral in identifying and evaluating unique pathways that may be exploited as therapeutic targets. Assessment of GEMM tumor burden on histopathological sections performed by manual inspection is both time consuming and prone to subjective bias. Therefore, an interplay of needs and challenges exists for computer aided detection tools, for the accurate and efficient analysis of these histopathology images. Our work demonstrates a simple machine learning approach called sparse principal component analysis (PCA) network, for automated detection of cancerous lesions on histological lung slides stained by hematoxylin and eosin (H&E). Methods: Our method comprises four steps: 1) cascaded sparse PCA; 2) graph-based PCA hashing; 3) block-wise histograms; and 4) support vector machine (SVM) classification. In our proposed architecture, sparse PCA is employed to learn the filter banks of the multiple stages. This is followed by a graph-based PCA hashing and block histograms for indexing and pooling. The meaningful features extracted from this sparse PCA are then fed to an SVM classifier. We tested the proposed sparse PCA network on H&E slides obtained from an inducible KrasG12D lung cancer mouse model. Our dataset consists of N = 21 whole slide histopathology lung images with 9 non-tumor bearing control mice and 12 mice with visible lung tumors. Tumor lesions from 12 lung images with visible tumors were visually identified by three trained individuals, which served as ground truth. The size of each image in our dataset is 2048 × 2048 pixels. Each image was divided into non-overlapping image patches of size 20 × 20 pixels consisting of a total of 12,361 cancer lesion patches and 207,839 non-cancer patches. We used 50% of the data for training and 50% of the data for testing our proposed sparse PCA network. We evaluated our algorithm using conventional metrics that have been used for evaluation of classification algorithms, namely precision (P), recall (R), and coverage measure (F-score). Results: The automatic cancer lesion detection results were compared with manually annotated ground truth. The proposed method achieves a cancer lesion detection accuracy of 97.98% with P = 0.8624, R = 0.9062 and F-score = 0.8790. The proposed method was found to take on average 17 minutes to train and learn a good representation for accurate and efficient classification of cancerous lesions within the images. Conclusion: We demonstrated a simple machine learning methodology for detection of cancerous lesions within histopathological lung images. Experimental results show that the proposed method is able to classify the regions of interest both efficiently and accurately. Future work will focus on feature extraction of individual tumors and tumor location within lungs. Citation Format: Sundaresh Ram, Wenfei Tang, Alexander J. Bell, Cara Spencer, Alexander Buschhuas, Charles R. Hatt, Marina P. di Magliano, Stefanie Galban, Craig J. Galban. Detection of cancer lesions in histopathological lung images using a sparse PCA network [abstract]. In: Proceedings of the AACR Virtual Special Conference on Artificial Intelligence, Diagnosis, and Imaging; 2021 Jan 13-14. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(5_Suppl):Abstract nr PO-086.

Chemoprotective effect of atorvastatin against benzo(a)pyrene-induced lung cancer via the inhibition of oxidative stress and inflammatory parameters.

BackgroundLung cancer affects approximately 9% of women and 17% of men worldwide, and has a mortality rate of 17%. Previously published studies have suggested that oxidative stress expansion can lead to lung cancer. The aim of the current study was to analyze the possible inhibitory pathway of atorvastatin against lung cancer cells in an in vivo model.MethodsThe cytotoxic effects of atorvastatin on lung cancer cell lines H460 and A549 were analyzed, as well as cell cycle arrest and cell morphology. Benzo(a)pyrene (BaP) was used for the induction of lung cancer in experimental rats, and atorvastatin (5, 10, and 20 mg/kg body weight) was used for treatment in a dose-dependent manner. Body weight and lung tumors were calculated at regular intervals. Antioxidants, pro-inflammatory cytokines, phase I and II antioxidant enzymes, polyamine enzymes, and apoptosis markers were determined at end of the experimental study.ResultsCell cycle arrest occurred at the G2/M phase after atorvastatin treatment. Atorvastatin increased cytochrome C expression and caspase activity in a dose-dependent manner, and increased the activity of antioxidative enzymes, such as GPx, SOD, GST, reduced glutathione, and catalase, and reduced the level of nitrate and LPO. It also altered the xanthine oxidase (XO), Lactic Acid Dehydrogenase (LDH), quinone reductase (QR), UDP-glucuronosyltransferase (UDP-GT), adenosine deaminase (ADA), Aryl hydrocarbon hydroxylase (AHH), 5'-nucleotidase, cytochrome P450, cytochrome B5 and NADPH cytochrome C reductase levels. Atorvastatin was found to modulate polyamine enzyme levels, such as histamine, spermine, spermidine, and putrescine, and significantly (P<0.001) reduced the pro-inflammatory cytokine levels, such as tumor necrosis factor-α. Interleukin (IL)-6 and interleukin-1β (IL-1β) increased caspase-3 and caspase-9 levels in a dose-dependent manner.ConclusionsOur findings indicate that atorvastatin can inhibit lung cancer through apoptosis.

SMOKING CARCINOGENS AND LUNG CANCER – A REVIEW

Smoking ambiguity contributes to a certain revelation regarding the process by which cancer is induced. In the laboratory, carcinogens induce clear lung tumor to lung cancer induction. For instance, carcinogenic chemicals, namely, 4(methyl nitrosomine)-1-(3-pyridyl)-1-butanol, and nitrosonornicotine (NNN) cause tumor malignancy. It is evident from the mechanistic studies that the carcinogens have a stronger tendency to mutate the genes like suppresser gene, a gene that encodes the receptor of the cell surface to the nucleus, thus, giving way to the proliferation of mutation leading to neoplastic cells. In this analysis article, we have discussed molecular mechanics that can cause cancer by nitrosamines such as nicotine-derived nitrosamine ketone and NNN regarding a variety of significant cigarette combustion carcinogens and the effort to introduce a different dimensional approach to the prevention of cancer, by understanding the perspective of various treatments.

Management of early lung tumorigenesis by corilagin via modulation of proliferating cell nuclear antigen and apoptotic pathway

Background: Corilagin is vastly found in plants and incorporated in countless traditional medicinal systems to treat various ailments including cancer disease. However, till now, no scientific study has been done to validate its efficiency with appropriate experimental evidence. Resulting, this study examined the ameliorative effects of corilagin in benzo(a)pyrene (B[a] P)-induced lung cancer in vivo. Methods: Experimentally, the B(a)P (50 mg/kg body weight [b.wt.]) was administered orally to initiate lung cancer in mice, and the supplementation of prophylactic and therapeutic treatment by corilagin (30 mg/kg b.wt.) was observed for changes in b.wt. and lung weight, tumor incidence, oxidative stress marker (lipid peroxidation), tumor markers (carcinoembryonic antigen, neuron-specific enolase, aryl hydrocarbon hydroxylase, lactate dehydrogenase, γ-glutamyl transpeptidase, 5′-nucleotidase, alpha-keto dehydrogenase, citric acid cycle enzymes isocitrate dehydrogenase, succinate dehydrogenase, and malate dehydrogenase), histological and immunohistochemical analysis (proliferating cell nuclear antigen), and apoptotic mRNA gene expression (Bax, Bcl-2, caspase-3, caspase-9, and cytochrome-c). Results: B(a)P-induced animals exhibited unusual changes in b.wt. and lung weight, tumor incidence, oxidative stress marker, enzymatic antioxidants, tumor markers, histological lung tissue damage, number of proliferating cell nuclear antigen, and apoptotic mRNA gene expression. On prophylactic and therapeutic treatment of corilagin, the animals showed reinstatement of all the above alterations to the near normal. Conclusion: These findings of all the analyzed data demonstrate that the B(a)P-induced animals of pre- and post-treated with corilagin may prevent initiation of lung cancer, hence establishing its chemopreventive effect against lung tumorigenesis.

An innovative NRF2 nano-modulator induces lung cancer ferroptosis and elicits an immunostimulatory tumor microenvironment.

Simultaneous targeting of both the tumor microenvironment and cancer cells by a single nanomedicine has not been reported to date. Here, we report the dual properties of zero-valent-iron nanoparticle (ZVI-NP) to induce cancer-specific cytotoxicity and anti-cancer immunity.Methods: Cancer-specific cytotoxicity induced by ZVI-NP was determined by MTT assay. Mitochondria functional assay, immunofluorescence staining, Western blot, RT-qPCR, and ChIP-qPCR assays were used to dissect the mechanism underlying ZVI-NP-induced ferroptotic cancer cell death. The therapeutic potential of ZVI-NP was evaluated in immunocompetent mice and humanized mice. Immune cell profiles of allografts and ex vivo cultured immune cells were examined by flow cytometry analysis, RT-qPCR assay, and immunofluorescence.Results: ZVI-NP caused mitochondria dysfunction, intracellular oxidative stress, and lipid peroxidation, leading to ferroptotic death of lung cancer cells. Degradation of NRF2 by GSK3/β-TrCP through AMPK/mTOR activation was enhanced in such cancer-specific ferroptosis. In addition, ZVI-NP attenuated self-renewal ability of cancer and downregulated angiogenesis-related genes. Importantly, ZVI-NP augmented anti-tumor immunity by shifting pro-tumor M2 macrophages to anti-tumor M1, decreasing the population of regulatory T cells, downregulating PD-1 and CTLA4 in CD8+ T cells to potentiate their cytolytic activity against cancer cells, while attenuating PD-L1 expression in cancer cells in vitro and in tumor-bearing immunocompetent mice. In particular, ZVI-NPs preferentially accumulated in tumor and lung tissues, leading to prominent suppression of tumor growth and metastasis.Conclusions: This dual-functional nanomedicine established an effective strategy to synergistically induce ferroptotic cancer cell death and reprogram the immunosuppressive microenvironment, which highlights the potential of ZVI-NP as an advanced integrated anti-cancer strategy.

Tachyplesin induces apoptosis in non-small cell lung cancer cells and enhances the chemosensitivity of A549/DDP cells to cisplatin by activating Fas and necroptosis pathway.

Cisplatin has strong broad-spectrum anticancer activity and is one of the most effective anticancer drugs currently used. The clinical application of cisplatin has led to the resistance of cancer cells to cisplatin. Tachyplesin is an active, natural marine peptide with antitumour activity. In the present study, we investigated whether tachyplesin can be used in non-small cell lung cancer (NSCLC) A549 and H460 cells as well as the cisplatin-resistant human A549/DDP NSCLC cell line. The results revealed that tachyplesin treatment significantly inhibited proliferation and induced apoptosis in A549 and H460 cells and the combination of tachyplesin and cisplatin significantly suppressed migration and improved sensitivity to cisplatin in A549/DDP cells. Further mechanistic examination revealed that tachyplesin induced apoptosis in A549/DDP cells by increasing Fas, FasL and p-RIPK1 levels. These results indicated that tachyplesin induces lung cancer death by activating the Fas, mitochondrial and necroptosis pathways. Tachyplesin could be developed as a candidate drug for cisplatin-resistant NSCLC.

Diallyl Sulfide-Mediated Modulation of the Fatty Acid Synthase (FASN) Leads to Cancer Cell Death in BaP-Induced Lung Carcinogenesis in Swiss Mice.

PurposeDiallyl sulfide (DAS), one of the organo-sulfur secondary metabolites in garlic, has been shown to inhibit the proliferation of cancer cells. The present study aimed to evaluate the mechanism of DAS in the prevention of benzo[a]pyrene (BaP)-induced lung cancer in a murine model.Materials and MethodsThe mice were exposed to 50 mg/kg of BaP twice a week for 4 weeks in order to induce lung carcinoma. Pretreatment of mice with DAS (100 mg/kg) was started 2 weeks before BaP exposure and further continued for 21 weeks. The effect of DAS and BaP was evaluated by studying various parameters in the serum and tissues of the treated or untreated BaP-exposed mice.ResultsThe histopathological findings demonstrated that DAS prevented the progression of malignant lung cancer and metastasis in the liver. A significant drop was observed in BaP-induced tumor marker enzymes (ADA, AHH, γ-GT, LDH) in the serum of the mice treated with DAS. Moreover, DAS treatment resulted in the recovery of antioxidant enzymes, SOD and CAT, in BaP-exposed mice. The induction of apoptosis and the destruction of cellular ROS were detected in cancer cells from the mice pre-treated with DAS. The immunohistochemical analysis revealed the up-regulation of fatty acid synthase (FASN) in the lungs and liver tissues of BaP-exposed mice and the treatment with DAS inhibited FASN expression.ConclusionThe findings of the present study indicated that DAS-induced apoptosis is strongly associated with the downregulation of FASN in tumor tissues. To the best of our knowledge, this is the first study that describes the role of FASN in BaP-induced lung carcinogenesis.

Immunosuppressive role of Benzo[a]pyrene in induction of lung cancer in mice

AimBenzo[a]pyrene [BP] is one of the major carcinogenic precursors of cigarette smoke that primary affects the lung at its first proximity. The goal of the current research was to elucidate new mechanisms underlying the tumorigenic impact of oral BP in the lung of mice, with focus on immunosuppressive effects and cancer stemming properties. MethodsFemale albino mice (n = 44) were divided into 2 groups: normal control and BP group. BP was administered orally to mice (50 mg/kg body weight), twice a week for four weeks in succession. At the end of experiment (22 weeks), gene expression were measured for transforming growth factor-β (TGF-β), cytotoxic T lymphocyte antigen-4 (CTLA-4), programmed death ligand 1(PD-L1), forkhead box protein P3 (FOXP3) and interleukin 12 (IL-12) and CD83+, CD8+ and CD166+ cell percentage were measured in lung tissue. ResultsThe results indicated the tumorigenic role of BP in the lung which was evidenced by histopathological examination. BP group also showed immunosuppressive role which evidenced by increased expression of lung TGF-β, CTLA-4, PD-L1, FOXP3 genes and decreased expression of lung IL-12 gene compared with normal control group. BP group also showed decreased CD83+ cells, CD8+ cells and increased number of CD166+ cells. ConclusionOur findings indicated that BP has immunosuppressive role in lung cancer besides increasing the percentage of cancer stem like cells.

Design and development of Tetrahydro-Quinoline derivatives as dual mTOR-C1/C2 inhibitors for the treatment of lung cancer

Lung cancer is one of the most prevailed cancer worldwide. Many genes get mutated in lung cancer but the involvement of EGFR, KRAS, PTEN and PIK3CA are more common. Unavailability of potent drugs and resistance to the available drugs are major concern in the treatment of lung cancer. In the present research, mTOR was selected as an important alternative target for the treatment of lung cancer which involves the PI3K/AKT/mTOR pathway. We studied binding interactions of AZD-2014 with the mTOR protein to identify important interactions required to design potent mTOR inhibitors which was supported by QSAR studies. Pharmacophore based virtual screening studies provided core scaffold, THQ. Based on molecular docking interactions, 31 THQ derivatives were synthesized and characterized. All compounds were screened for cellular mTOR enzyme assay along with antiproliferative activity against the panel of cancerous cell lines, from which 6 compounds were further screened for colony forming assay. Two most potent compounds, HB-UC-1 and HB-UC-5, were further screened for flow cytometry analysis, gene expression study and western blot analysis. Gene expression study revealed the efficiency of compound HB-UC-1 against both mTORC1 and mTORC2 by affecting downstream regulators of mTORC1 (E4BP4, eIF4EBP1) and mTORC2 (PCK1), respectively. In western blot analysis, both compounds, inhibited phosphorylation of AKT S473 which proved the efficiency these compounds against the mTORC2. These two compounds were further screened for in-vivo biological evaluation. Both compounds increased lifespan of cancer-bearing animals with improvement in mean survival time. Further, in bezopyrene induced lung cancer animal model, both compounds showed effectiveness through the biochemical parameters and histopathological evaluation of the lung tissue. In future, potent hit compound from this series could be modified to develop lead mTOR inhibitors for the treatment of lung cancer.

Raddeanin A suppresses lung cancer cell proliferation via induction of apoptosis and increased production of ROS

At present, in vitro cell experiments have confirmed that RaddeaninA can effectively inhibit the proliferation of some tumor cells, but the effect of RaddeaninA on lung cancer cells has not been observed. Therefore, this study explored its effect on lung cancer cells and its mechanism of action. Human lung cancer cell lines were treated with serum-free medium and varied concentrations of Raddeanin A. Cell proliferation and apoptosis were determined using MTT, and flow cytometric assays, respectively. The intracellular level of ROS was determined using DCFH-DA assay. Protein and mRNA expressions of bax, bcl-2 and cyt c were measured using Western blotting and qRT-PCR. RaddeaninA treatment can promote PC-9 cell apoptosis in a time and dose-dependent manner (p&lt;0.05). Treatment of PC-9 cells with Raddeanin significantly and dose-dependently increased the activities of caspase-9 and caspase-3 (p&lt;0.05), and led to significant and dose-dependent increases in ROS levels (p&lt;0.05). Treatment of PC-9 cells with Raddeanin A led to significant and dose-dependent decreases in mitochondrial membrane potential (p&lt;0.05). It significantly and dose-dependently upregulated bax mRNA and protein expressions, but down-regulated bcl-2 mRNA and protein expressions significantly and dose-dependently (p&lt;0.05). On the other hand, Raddeanin significantly and dose-dependently down-regulated cytoplasmic bax protein expression, while upregulating cyt c expression (p&lt;0.05). Similarly, bax protein expression was significantly and dose-dependently upregulated in mitochondria, but the corresponding cyt c expression was significantly and dose-dependently down-regulated (p&lt;0.05). Raddeanin A is a potential and effective lung cancer chemotherapy drug, which can induce lung cancer cell apoptosis and inhibit proliferation.

Characterization of a Novel Barbituric Acid and Two Thiobarbituric Acid Compounds for Lung Cancer Treatment.

Previously, we reported the identification of a cytotoxic chemotype compound CC-I (1a), a derivative of thiobarbituric acid. We also reported the anticancer activity of a series of novel thio- and seleno-barbituric acid analogs. We herein evaluated the effect of 1a and its modified compounds on in vitro and in vivo lung cancer models. The compounds 1b and 2a showed more potent cytotoxicity than 1a to lung cancer cells. Moreover, 1b did not have any cytotoxicity on normal cells, such as fibroblasts. In the human lung cancer A549 mouse tumor xenograft model, 1b and 2a showed more pronounced antitumor effects than 1a In the A549 lung cancer cells, 1a induced cell death mainly via JNK and p38 MAPK activation. However, compound 1b and 2a induced lung cancer cell death mostly through JNK activation. The results suggest that 1b and 2a can be useful therapeutic agents for lung cancer.

Poly(ADP-ribose) glycohydrolase silencing-mediated H2B expression inhibits benzo(a)pyrene-induced carcinogenesis.

Poly(ADP‐ribose) glycohydrolase (PARG) as a main enzyme hydrolyzing poly(ADP‐ribose) in eukaryotes, and its silencing can inhibit benzo(a)pyrene (BaP)‐induced carcinogenesis. A thorough understanding of the mechanism of PARG silenced inhibition of BaP‐induced carcinogenesis provides a new therapeutic target for the prevention and treatment of environmental hazard induced lung cancer. We found that the expression of several subtypes of the histone H2B was downregulated in BaP‐induced carcinogenesis via PARG silencing as determined by label‐free proteomics and confirmed by previous cell line‐ and mouse model‐based studies. Analysis using the GEPIA2 online tool indicated that the transcription levels of H2BFS, HIST1H2BD, and HIST1H2BK in lung adenocarcinoma (LUAD) tissues and squamous cell lung carcinoma (LUSC) tissues were higher than those in normal lung tissues, while the transcription levels of HIST1H2BH in LUSC tissues were higher than those in normal lung tissues. The expression levels of HIST1H2BB, HIST1H2BH, and HIST1H2BL were significantly different in different lung cancer (LC) stages. Moreover, the expression of H2BFS, HIST1H2BD, HIST1H2BJ, HIST1H2BK, HIST1H2BL, HIST1H2BO, HIST2H2BE, and HIST2H2BF was positively correlated with that of PARG in LC tissues. Analysis of the Kaplan‐Meier plotter database indicated that high H2B levels predicted low survival in all LC patients suggesting that H2B could be a new biomarker for determining the prognosis of the LC, and that its expression can be inhibited by PARG silencing in BaP‐induced carcinogenesis.

IL-27 along with IL-28B ameliorates the pulmonary redox impairment, inflammation and immunosuppression in benzo(a)pyrene induced lung cancer bearing mice

AimsThe major cause behind lung cancer development is exposure to various polycyclic aromatic hydrocarbons like benzo(a)pyrene (BaP) present in tobacco smoke, motor vehicle, and industrial exhaust. BaP is reported to induce the expression of various pro-inflammatory cytokines and matrix remodeling proteins. It is also responsible for dysfunction and exhaustion of the killing capacity of CD8+ T lymphocytes, one of the important components of the immune system which can kill tumor cells. We tried to evaluate the synergistic role of IL-27 and IL-28B in modulation of BaP-induced lung carcinogenesis associated with various hallmarks like pulmonary redox imbalance, angiogenesis, inflammation and cell proliferation in lung tissue. Main methodBaP was treated to Swiss albino mice to develop lung tumor. After the confirmation of lung tumor development Swiss albino mice were treated with IL-27 and IL-28B alone or in combination intraperitoneally. Histological analysis, immunohistochemistry, biochemical assay, western blot analysis, cell cytotoxicity assay, real-time PCR assay etc. were performed to evaluate the modulatory role of IL-27 and IL-28B. Key findingsWe observed that IL-27 and IL-28B were able to suppress the expression of lung cancer-associated NFkB, COX-2, and iNOS. The expression of TNF-α, PCNA and some matrix remodeling enzymes were also modulated upon IL-27 and IL-28B treatment. Although the population of lung residing CD8+ T cells in tumor bearing lung tissue were unresponsive but the activity of systemic CD8+ cells was increased. SignificanceResults hinted that IL-27 along with IL-28B were able to ameliorate various hallmarks ranging from angiogenesis to inflammation associated with the BaP-induced lung carcinogenesis. From this study, we propose that IL-27 and IL28B can be used as immunotherapeutic agent to regulate lung cancer.