Tempol differently affects cellular redox changes and antioxidant enzymes in various lung-related cells

AbstractTempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl) is a potential redox agent in cells. The present study investigated changes in cellular reactive oxygen species (ROS) and glutathione (GSH) levels and in antioxidant enzymes, in Tempol-treated Calu-6 and A549 lung cancer cells, normal lung WI-38 VA-13 cells, and primary pulmonary fibroblasts. Results demonstrated that Tempol (0.5–4 mM) either increased or decreased general ROS levels in lung cancer and normal cells at 48 h and specifically increased O2•− levels in these cells. In addition, Tempol differentially altered the expression and activity of antioxidant enzymes such as superoxide dismutase, catalase, and thioredoxin reductase1 (TrxR1) in A549, Calu-6, and WI-38 VA-13 cells. In particular, Tempol treatment increased TrxR1 protein levels in these cells. Tempol at 1 mM inhibited the growth of lung cancer and normal cells by about 50% at 48 h but also significantly induced cell death, as evidenced by annexin V-positive cells. Furthermore, down-regulation of TrxR1 by siRNA had some effect on ROS levels as well as cell growth inhibition and death in Tempol-treated or -untreated lung cells. In addition, some doses of Tempol significantly increased the numbers of GSH-depleted cells in both cancer cells and normal cells at 48 h. In conclusion, Tempol differentially increased or decreased levels of ROS and various antioxidant enzymes in lung cancer and normal cells, and induced growth inhibition and death in all lung cells along with an increase in O2•− levels and GSH depletion.

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Tempol Differently Affects Cellular Redox Status and Antioxidant Enzymes in Various Lung-related Cells

10.21203/rs.3.rs-257487/v1 ◽

2021 ◽

Author(s):

woo hyun park

Keyword(s):

Lung Cancer ◽

Antioxidant Enzymes ◽

Growth Inhibition ◽

Cancer Cells ◽

Annexin V ◽

Redox Status ◽

Normal Lung ◽

Normal Cells ◽

Cellular Redox ◽

Protein Levels

Abstract Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl) is a potential redox agent in cells. The present study investigated changes in cellular redox status [reactive oxygen species (ROS) and glutathione (GSH) levels] and in antioxidant enzymes, in Tempol-treated Calu-6 and A549 lung cancer cells, normal lung WI-38 VA-13 cells, and primary pulmonary fibroblasts. Results demonstrated that Tempol (0.5 ~ 4 mM) either increased or decreased general ROS levels in lung cancer and normal cells at 48 h and specifically increased O2•− levels in these cells. In addition, Tempol differentially altered the expression and activity of superoxide dismutase, catalase, and thioredoxin reductase1 (TrxR1) in A549, Calu-6, and WI-38 VA-13 cells. In particular, Tempol increased TrxR1 protein levels in these cells. Tempol at 1 mM inhibited the growth of lung cancer and normal cells by about 50% at 48 h but also significantly induced cell death, as evidenced by annexin V-positive cells. Furthermore, down-regulation of TrxR1 by siRNA somewhat affected the levels of cell growth inhibition and death as well as ROS in Tempol-treated cells. In addition, Tempol increased the numbers of GSH-depleted cells in both cancer cells and normal cells at 48h. In conclusion, Tempol differentially increased or decreased levels of ROS and various antioxidant enzymes in lung cancer and normal cells, and induced growth inhibition and death in all lung cells along with an increase in O2•− levels and GSH depletion.

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Glycosylated and non-glycosylated quantum dot-displayed peptides trafficked indiscriminately inside lung cancer cells but discriminately sorted in normal lung cells: An indispensable part in nanoparticle-based intracellular drug delivery

Asian Journal of Pharmaceutical Sciences ◽

10.1016/j.ajps.2017.12.002 ◽

2018 ◽

Vol 13 (3) ◽

pp. 197-211 ◽

Cited By ~ 3

Author(s):

Roger Salvacion Tan

Keyword(s):

Lung Cancer ◽

Drug Delivery ◽

Quantum Dot ◽

Cancer Cells ◽

Lung Cancer Cells ◽

Normal Lung ◽

Lung Cells ◽

Intracellular Drug Delivery

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Functional polyesters enable selective siRNA delivery to lung cancer over matched normal cells

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1606886113 ◽

2016 ◽

Vol 113 (39) ◽

pp. E5702-E5710 ◽

Cited By ~ 42

Author(s):

Yunfeng Yan ◽

Li Liu ◽

Hu Xiong ◽

Jason B. Miller ◽

Kejin Zhou ◽

...

Keyword(s):

Lung Cancer ◽

Side Effects ◽

Cancer Cells ◽

Drug Carrier ◽

Sirna Delivery ◽

Normal Lung ◽

Cancer Therapies ◽

Normal Cells ◽

Alternative Approach ◽

Dividing Cells

Conventional chemotherapeutics nonselectively kill all rapidly dividing cells, which produces numerous side effects. To address this challenge, we report the discovery of functional polyesters that are capable of delivering siRNA drugs selectively to lung cancer cells and not to normal lung cells. Selective polyplex nanoparticles (NPs) were identified by high-throughput library screening on a unique pair of matched cancer/normal cell lines obtained from a single patient. Selective NPs promoted rapid endocytosis into HCC4017 cancer cells, but were arrested at the membrane of HBEC30-KT normal cells during the initial transfection period. When injected into tumor xenografts in mice, cancer-selective NPs were retained in tumors for over 1 wk, whereas nonselective NPs were cleared within hours. This translated to improved siRNA-mediated cancer cell apoptosis and significant suppression of tumor growth. Selective NPs were also able to mediate gene silencing in xenograft and orthotopic tumors via i.v. injection or aerosol inhalation, respectively. Importantly, this work highlights that different cells respond differentially to the same drug carrier, an important factor that should be considered in the design and evaluation of all NP carriers. Because no targeting ligands are required, these functional polyester NPs provide an exciting alternative approach for selective drug delivery to tumor cells that may improve efficacy and reduce adverse side effects of cancer therapies.

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Differential effect of hypoxia and acidity on lung cancer cell and fibroblast metabolism

Biochemistry and Cell Biology ◽

10.1139/bcb-2016-0197 ◽

2017 ◽

Vol 95 (3) ◽

pp. 428-436 ◽

Cited By ~ 7

Author(s):

Alexandra Giatromanolaki ◽

Maria Liousia ◽

Stella Arelaki ◽

Dimitra Kalamida ◽

Stamatia Pouliliou ◽

...

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Metabolic Response ◽

Expression Patterns ◽

Lung Fibroblasts ◽

Normal Lung ◽

Lung Cancer Cell ◽

Protein Levels

This study examined the metabolic response of lung cancer cells and normal lung fibroblasts to hypoxia and acidity. GLUT1 and HXKII mRNA/protein expression was up-regulated under hypoxia in the MRC5 fibroblasts and in the A549 and H1299 lung cancer cell lines, indicating intensified glucose absorption and glycolysis. Under hypoxia, the LDHA mRNA and LDH5 protein levels increased in the cancer cells but not in the fibroblasts. Acidity suppressed the above-mentioned hypoxia effect. PDH-kinase-1 (PDK1 mRNA and protein) and inactive phosphorylated-PDH protein levels were induced under hypoxia in the cancer cells, whereas these were reduced in the MRC5 lung fibroblasts. In human tissue sections, the prevalent expression patterns supported the contrasting metabolic behavior of cancer cells vs. tumor fibroblasts. The monocarboxylate/lactate transporter 1 (MCT1) was up-regulated in all the cell lines under hypoxic conditions, but it was suppressed under acidic conditions. The mitochondrial DNA (mtDNA) content per cell decreased significantly in the A549 cancer cell line under hypoxia, but it increased in the MRC5 fibroblasts. Taking into account these findings, we suggest that, under hypoxia, cancer cells intensify the anaerobic direction in glycolysis, while normal fibroblasts prefer to seek energy by intensifying the aerobic use of the available oxygen.

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The therapeutic effects of vitamin C on lung cancer cells.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.e18523 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. e18523-e18523

Author(s):

Ashorne Krithiesh Mahenthiran ◽

Gurusingham Sitta Sittampalam ◽

Raj Somasundaram ◽

Sanjit Nirmalanandhan

Keyword(s):

Lung Cancer ◽

Vitamin C ◽

Cancer Cells ◽

Cell Lines ◽

In Vitro Study ◽

Lung Cancer Cells ◽

Normal Lung ◽

Lung Cells ◽

Experimental Conditions

e18523 Background: In this in-vitro study, we determined the effects of vitamin C (Ascorbic acid), an essential vitamin, on two different lung cancer cell lines (H358 – Bronchioalveolar Carcinoma and A549 – Epithelial Lung carcinoma) and two normal lung cell lines as control groups (MRC5 – Human lung fibroblast tissue and NL20 – Lung epithelial cells). Methods: In the study, the four cell lines were treated with Vitamin C starting from 0.005 molar concentrations and serially diluted down 1:3 ratios to low nM concentrations. All experiments were carried out in a period of 4 weeks. The viability of the cell lines after the drug treatment was measured using a MTS cell proliferation assay. Results: The study was inconclusive since the viability of both normal and lung cells were equally affected under the experimental conditions except that the dosage of vitamin C that killed 50% of H358 was at a slightly lower concentration than the dosage of vitamin C that killed 50% of the normal lung cells. These results show that there is a possibility of an optimal dosage that will only harm cancerous cells in specific cancers and not on all cancers. Conclusions: These results were inconclusive; probably due to the fact that experimental conditions in this in-vitro study may not be appropriate to show the effects of Vitamin C on lung cancer cells. It is possible that lower dosages of vitamin C may still kill cancer cells selectively, and may also be more effective in cancers in ther tissues. Despite these drawbacks, in-vivo experiments in animal models with lung cancer may show the benefits of Vitamin C in combination with standard of care cancer drugs. Future experiments will examine combinations experiments in vitro and in animals to study the beneficial effects of Vitamin C.

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Differential regulation of rho GTPases during lung adenocarcinoma migration and invasion reveals a novel role of the tumor suppressor StarD13 in invadopodia regulation

Cell Communication and Signaling ◽

10.1186/s12964-020-00635-5 ◽

2020 ◽

Vol 18 (1) ◽

Cited By ~ 3

Author(s):

Maria Al Haddad ◽

Rayane El-Rif ◽

Samer Hanna ◽

Leila Jaafar ◽

Rayanne Dennaoui ◽

...

Keyword(s):

Lung Cancer ◽

Cell Migration ◽

Tumor Suppressor ◽

Cancer Cells ◽

Rho Gtpases ◽

Normal Lung ◽

Migration And Invasion ◽

Lung Cells ◽

Invadopodia Formation

Abstract Background Lung cancer is the second most commonly occurring cancer. The ability to metastasize and spread to distant locations renders the tumor more aggressive. Members of the Rho subfamily of small GTP-binding proteins (GTPases) play a central role in the regulation of the actin cytoskeleton and in cancer cell migration and metastasis. In this study we investigated the role of the RhoA/Cdc42 GAP, StarD13, a previously described tumor suppressor, in malignancy, migration and invasion of the lung cancer cells A549. Methods We knocked down StarD13 expression in A549 lung cancer cells and tested the effect on cell migration and invadopodia formation using time lapse imaging and invasion assays. We also performed rescue experiments to determine the signaling pathways downstream of StarD13 and transfected the cells with FRET biosensors for RhoGTPases to identify the proteins involved in invadopodia formation. Results We observed a decrease in the level of expression of StarD13 in lung tumor tissues compared to normal lung tissues through immunohistochemistry. StarD13 also showed a lower expression in the lung adenocarcinoma cell line A549 compared to normal lung cells, WI38. In addition, the depletion of StarD13 increased cell proliferation and viability in WI38 and A549 cells, suggesting that StarD13 might potentially be a tumor suppressor in lung cancer. The depletion of StarD13, however, inhibited cell motility, conversely demonstrating a positive regulatory role in cell migration. This was potentially due to the constitutive activation of RhoA detected by pull down and FRET assays. Surprisingly, StarD13 suppressed cell invasion by inhibiting Cdc42-mediated invadopodia formation. Indeed, TKS4 staining and invadopodia assay revealed that StarD13 depletion increased Cdc42 activation as well as invadopodia formation and matrix degradation. Normal lung cells depleted of StarD13 also produced invadopodia, otherwise a unique hallmark of invasive cancer cells. Cdc42 knock down mimicked the effects of StarD13, while overexpression of a constitutively active Cdc42 mimicked the effects of its depletion. Finally, immunostaining and FRET analysis revealed the absence of StarD13 in invadopodia as compared to Cdc42, which was activated in invadopodia at the sites of matrix degradation. Conclusion In conclusion, StarD13 plays distinct roles in lung cancer cell migration and invasion through its differential regulation of Rho GTPases.

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Induction of p27KIP1 as a Mechanism Underlying NS398-Induced Growth Inhibition in Human Lung Cancer Cells

Molecular Pharmacology ◽

10.1124/mol.58.6.1398 ◽

2000 ◽

Vol 58 (6) ◽

pp. 1398-1403 ◽

Cited By ~ 54

Author(s):

Wen-Chun Hung ◽

Hui-Chiu Chang ◽

Mei-Ren Pan ◽

Te-Hsiu Lee ◽

Lea-Yea Chuang

Keyword(s):

Lung Cancer ◽

Growth Inhibition ◽

Cancer Cells ◽

Human Lung ◽

Lung Cancer Cells ◽

Human Lung Cancer ◽

Human Lung Cancer Cells

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Sestrin2 Expression Has Regulatory Properties and Prognostic Value in Lung Cancer

Journal of Personalized Medicine ◽

10.3390/jpm10030109 ◽

2020 ◽

Vol 10 (3) ◽

pp. 109 ◽

Cited By ~ 1

Author(s):

Hee Sung Chae ◽

Minchan Gil ◽

Subbroto Kumar Saha ◽

Hee Jeung Kwak ◽

Hwan-Woo Park ◽

...

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Prognostic Value ◽

Mammalian Target Of Rapamycin ◽

Cancer Cell Line ◽

Lung Cancer Cell Line ◽

Lung Cancer Cells ◽

Normal Lung ◽

Therapeutic Modalities

Lung cancer remains the most dangerous type of cancer despite recent progress in therapeutic modalities. Development of prognostic markers and therapeutic targets is necessary to enhance lung cancer patient survival. Sestrin family genes (Sestrin1, Sestrin2, and Sestrin3) are involved in protecting cells from stress. In particular, Sestrin2, which mainly protects cells from oxidative stress and acts as a leucine sensor protein in mammalian target of rapamycin (mTOR) signaling, is thought to affect various cancers in different ways. To investigate the role of Sestrin2 expression in lung cancer cells, we knocked down Sestrin2 in A549, a non-small cell lung cancer cell line; this resulted in reduced cell proliferation, migration, sphere formation, and drug resistance, suggesting that Sestrin2 is closely related to lung cancer progression. We analyzed Sestrin2 expression in human tissue using various bioinformatic databases and confirmed higher expression of Sestrin2 in lung cancer cells than in normal lung cells using Oncomine and the Human Protein Atlas. Moreover, analyses using Prognoscan and KMplotter showed that Sestrin2 expression is negatively correlated with the survival of lung cancer patients in multiple datasets. Co-expressed gene analysis revealed Sestrin2-regulated genes and possible associated pathways. Overall, these data suggest that Sestrin2 expression has prognostic value and that it is a possible therapeutic target in lung cancer.

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Lung Cancer Detection Using Marker Controlled Watershed with SVM

GUB Journal of Science and Engineering ◽

10.3329/gubjse.v5i1.47897 ◽

2018 ◽

Vol 5 (1) ◽

pp. 24-30

Author(s):

Fatema Tuj Johora ◽

Mehdi Hassan Jony ◽

Md Shakhawat Hossain ◽

Humayun Kabir Rana

Keyword(s):

Lung Cancer ◽

Cancer Cells ◽

Cancer Detection ◽

Early Stage ◽

Support Vector ◽

Lung Cells ◽

Science And Engineering ◽

Efficient Performance ◽

Occurrence Matrix ◽

Lung Cancer Detection

Lung cancer is one of the most dangerous diseases and prediction of it, is the most challenging problem nowadays. Most of the cancer cells are overlapped with each other. It is hard to detect the cells but also essential to identify the presence of cancer cells in the early stage. Early detection of lung cancer may reduce the death rate. In this study, we used the Grey Level Co-occurrence Matrix (GLCM) to extract the feature of cancer affected lung image and then Support Vector Machine (SVM) has been used to detect normal and abnormal lung cells after implementing the features. Our experimental evaluation using MATLAB demonstrates the efficient performance of the proposed system and in the result. GUB JOURNAL OF SCIENCE AND ENGINEERING, Vol 5(1), Dec 2018 P 24-30

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