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  1. Article ; Online: Pathologic Processing of Lung Cancer Resection Specimens After Neoadjuvant Therapy.

    Weissferdt, Annikka / Leung, Cheuk H / Lin, Heather / Sepesi, Boris / William, William N / Swisher, Stephen G / Cascone, Tina / Lee, J Jack / Pataer, Abujiang

    Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

    2023  Volume 37, Issue 1, Page(s) 100353

    Abstract: Neoadjuvant treatment of non-small cell lung cancer challenges the traditional processing of pathology specimens. Induction therapy before resection allows evaluation of the efficacy of neoadjuvant agents at the time of surgery. Many clinical trials use ... ...

    Abstract Neoadjuvant treatment of non-small cell lung cancer challenges the traditional processing of pathology specimens. Induction therapy before resection allows evaluation of the efficacy of neoadjuvant agents at the time of surgery. Many clinical trials use pathologic tumor response, measured as major pathologic response (MPR, ≤10% residual viable tumor [RVT]) or complete pathologic response (CPR, 0% RVT) as a surrogate of clinical efficacy. Consequently, accurate pathologic evaluation of RVT is crucial. However, pathologic assessment has not been uniform, which is particularly true for sampling of the primary tumor, which instead of the traditional processing, requires different tissue submission because the focus has shifted from tumor typing alone to RVT scoring. Using a simulation study, we analyzed the accuracy rates of %RVT, MPR, and CPR of 31 pretreated primary lung tumors using traditional grossing compared with the gold standard of submitting the entire residual primary tumor and identified the minimum number of tumor sections to be submitted to ensure the most accurate scoring of %RVT, MPR, and CPR. Accurate %RVT, MPR, and CPR calls were achieved in 52%, 87%, and 81% of cases, respectively, using the traditional grossing method. Accuracy rates of at least 90% for these parameters require either submission of all residual primary tumor or at least 20 tumor sections. Accurate %RVT, MPR, and CPR scores cannot be achieved with traditional tumor grossing. Submission of the entire primary tumor, up to a maximum of 20 sections, is required for the most accurate reads.
    MeSH term(s) Humans ; Lung Neoplasms/surgery ; Lung Neoplasms/drug therapy ; Carcinoma, Non-Small-Cell Lung/surgery ; Carcinoma, Non-Small-Cell Lung/drug therapy ; Neoadjuvant Therapy/methods ; Lung/pathology ; Treatment Outcome
    Language English
    Publishing date 2023-10-14
    Publishing country United States
    Document type Journal Article
    ZDB-ID 645073-8
    ISSN 1530-0285 ; 0893-3952
    ISSN (online) 1530-0285
    ISSN 0893-3952
    DOI 10.1016/j.modpat.2023.100353
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Inhibition of RNA-dependent protein kinase (PKR) leads to cancer cell death and increases chemosensitivity.

    Pataer, Abujiang / Swisher, Stephen G / Roth, Jack A / Logothetis, Christopher J / Corn, Paul G

    Cancer biology & therapy

    2009  Volume 8, Issue 3, Page(s) 245–252

    Abstract: RNA-dependent protein kinase is an interferon-induced, double-stranded (ds), RNA-activated serine/threonine protein kinase involved in the eukaryotic response to viral infection. While PKR also functions in cellular differentiation, growth control and ... ...

    Abstract RNA-dependent protein kinase is an interferon-induced, double-stranded (ds), RNA-activated serine/threonine protein kinase involved in the eukaryotic response to viral infection. While PKR also functions in cellular differentiation, growth control and apoptosis, its role in human cancer remains poorly understood. To explore a role for PKR in human cancer, we evaluated PKR expression and function in a series of cancer cell lines from different tumor types. We observed that PKR protein expression is high in various cancer cells and low in normal cells. Knockdown of PKR protein expression by PKR siRNA induced cell death, indicating a PKR-dependent survival pathway under normal growth conditions. Inhibition of PKR signaling using a dominant negative adenoviral PKR mutant (Ad-Delta6PKR) also induced cancer cell apoptosis via a mechanism that blocks activation of AKT-mediated survival while simultaneously inducing ER stress. ER stress-mediated apoptosis was evidenced by unregulated expression of phosphorylated JNK (p-JNK), phosphorylated cJun (p-cJun), and caspase-4 and was significantly reduced in cancer cells treated with JNK and caspase-4 inhibitors. We further demonstrated that inhibition of PKR signaling via either siRNA or Ad-Delta6PKR sensitizes cancer cells to etoposide or cisplatin-mediated cell death. Our results suggest a rationale to develop therapeutic strategies that target PKR signaling in human cancer cells.
    MeSH term(s) Antineoplastic Agents, Phytogenic/pharmacology ; Apoptosis/drug effects ; Caspases, Initiator/genetics ; Caspases, Initiator/metabolism ; Cell Line, Tumor ; Cisplatin/pharmacology ; Drug Resistance, Neoplasm ; Endoplasmic Reticulum/physiology ; Etoposide/pharmacology ; Female ; Gene Deletion ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Humans ; JNK Mitogen-Activated Protein Kinases/genetics ; JNK Mitogen-Activated Protein Kinases/metabolism ; Male ; Neoplasms/drug therapy ; Neoplasms/genetics ; Neoplasms/metabolism ; Signal Transduction ; Stress, Physiological ; eIF-2 Kinase/biosynthesis ; eIF-2 Kinase/genetics
    Chemical Substances Antineoplastic Agents, Phytogenic ; Etoposide (6PLQ3CP4P3) ; eIF-2 Kinase (EC 2.7.11.1) ; JNK Mitogen-Activated Protein Kinases (EC 2.7.11.24) ; CASP4 protein, human (EC 3.4.22.-) ; Caspases, Initiator (EC 3.4.22.-) ; Cisplatin (Q20Q21Q62J)
    Language English
    Publishing date 2009-02-08
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2146305-0
    ISSN 1555-8576 ; 1538-4047
    ISSN (online) 1555-8576
    ISSN 1538-4047
    DOI 10.4161/cbt.8.3.7386
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article: Genetically targeted cancer therapy: tumor destruction by PKR activation.

    Vorburger, Stephan A / Pataer, Abujiang / Swisher, Stephen G / Hunt, Kelly K

    American journal of pharmacogenomics : genomics-related research in drug development and clinical practice

    2004  Volume 4, Issue 3, Page(s) 189–198

    Abstract: The is a double-stranded RNA-activated protein kinase (PKR) has been largely investigated for its key role in viral host defense. Although best characterized by its function in mediating the antiviral and antiproliferative effects of interferon (IFN), ... ...

    Abstract The is a double-stranded RNA-activated protein kinase (PKR) has been largely investigated for its key role in viral host defense. Although best characterized by its function in mediating the antiviral and antiproliferative effects of interferon (IFN), PKR is also implicated in transcriptional regulation, cell differentiation, signal transduction, and tumor suppression. However, recent findings identifying PKR as an important effector of apoptosis have led to an increased interest in PKR modulation as an antitumor strategy. PKR can either be up-regulated through direct induction by the transcription factor E2F-1, or it can be activated through direct protein-protein interactions with the melanoma differentiation-associated gene-7 (MDA7, IL-24). Additionally, the intracellular formation of double-stranded RNA by transfection with antisense RNA complementary to tumor-specific RNA sequences can induce PKR activation and apoptosis selective to these tumor cells. The growing application of viral vector-based gene therapies and oncolytic, replicating viruses that must elude viral defense in order to be effective, has also drawn attention to PKR. Oncolytic viruses, like the attenuated herpes simplex virus R3616, the vesicular stomatitis virus, or reovirus, specifically replicate in tumor cells only because the viral host defense in the permissive cells is suppressed. In this article we review the role of PKR as an effector of apoptosis and a target for tumor treatment strategies and discuss the potential of PKR-modifying agents to treat patients with cancer. Targeted gene therapy against cancer can be approached by activation of PKR with the down-regulation of protein synthesis and induction of apoptosis, or by suppression of PKR with the propagation of oncolytic virus. Since the PKR pathway can be modified by many routes, antitumor therapies combining oncolytic virus, gene therapies, and chemotherapy with PKR modifiers are likely to emerge in the near future as therapeutic options in the treatment of patients with cancer.
    MeSH term(s) Animals ; Apoptosis/physiology ; Enzyme Activation/genetics ; Enzyme Inhibitors/therapeutic use ; Genetic Therapy ; Humans ; Neoplasms/genetics ; Neoplasms/therapy ; eIF-2 Kinase/antagonists & inhibitors ; eIF-2 Kinase/genetics ; eIF-2 Kinase/metabolism
    Chemical Substances Enzyme Inhibitors ; eIF-2 Kinase (EC 2.7.11.1)
    Language English
    Publishing date 2004-05-06
    Publishing country New Zealand
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, U.S. Gov't, P.H.S. ; Review
    ZDB-ID 2060524-9
    ISSN 1175-2203
    ISSN 1175-2203
    DOI 10.2165/00129785-200404030-00006
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Development of Ad-mda7/IL-24-resistant lung cancer cell lines.

    Pataer, Abujiang / Chada, Sunil / Roth, Jack A / Hunt, Kelly K / Swisher, Stephen G

    Cancer biology & therapy

    2007  Volume 7, Issue 1, Page(s) 103–108

    Abstract: Many cancers can become resistant to repeated administration of even the most effective therapeutic agents. In developing adenoviral mda-7/IL-24 (Ad-mda-7/IL-24) therapy for lung cancer, we have anticipated this potential clinical problem by attempting ... ...

    Abstract Many cancers can become resistant to repeated administration of even the most effective therapeutic agents. In developing adenoviral mda-7/IL-24 (Ad-mda-7/IL-24) therapy for lung cancer, we have anticipated this potential clinical problem by attempting to identify the molecular mechanisms of Ad-mda7/IL-24 resistance in several Ad-mda7/IL-24-resistant lung cancer cell lines that we have developed. For the present study, we established four Admda7- resistant cell lines by repeated selection of resistant clones of parental Ad-mda7-sensitive A549 cells: two lines (A549R1 and A549R2) resistant to both adenoviral vector and the mda-7 gene and two (A549R3 and A549R4) resistant to the therapeutic mda-7 gene only. As shown by western blot analysis of several known anti-apoptotic proteins, parental A549 and resistant A549R3 cells expressed similar levels of AKT and phosphorylated AKT (p-AKT), whereas resistant A549R3 and A549R4 cells expressed higher levels of bcl-2 and lower levels of bcl-xL than did their parental cells. As shown by flow-cytometric analysis, treating resistant A549R3 and A549R4 cells with a combination of Ad-mda7 and 17-allyl-amino-17-demethoxygeldanamycin (17AAG) (50 nM) for 48 hours enhanced apoptosis. Together, these in vitro findings indicate that an antiapoptotic mechanism may underlie Ad-mda7 resistance and that such resistance can be overcome by addition of 17AAG. Further investigations along these lines are warranted.
    MeSH term(s) Adenoviridae/genetics ; Benzoquinones/therapeutic use ; Cell Line, Tumor ; Coxsackie and Adenovirus Receptor-Like Membrane Protein ; Genetic Therapy ; Humans ; Interleukins/analysis ; Interleukins/genetics ; Lactams, Macrocyclic/therapeutic use ; Lung Neoplasms/therapy ; Proto-Oncogene Proteins c-akt/analysis ; Receptors, Virus/analysis
    Chemical Substances Benzoquinones ; CLMP protein, human ; Coxsackie and Adenovirus Receptor-Like Membrane Protein ; Interleukins ; Lactams, Macrocyclic ; Receptors, Virus ; interleukin-24 ; tanespimycin (4GY0AVT3L4) ; Proto-Oncogene Proteins c-akt (EC 2.7.11.1)
    Language English
    Publishing date 2007-10-13
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2146305-0
    ISSN 1555-8576 ; 1538-4047
    ISSN (online) 1555-8576
    ISSN 1538-4047
    DOI 10.4161/cbt.7.1.5162
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Structural mapping of post-translational modifications in human interleukin-24: role of N-linked glycosylation and disulfide bonds in secretion and activity.

    Fuson, Kerry L / Zheng, Mingzhong / Craxton, Molly / Pataer, Abujiang / Ramesh, Rajagopal / Chada, Sunil / Sutton, R Bryan

    The Journal of biological chemistry

    2009  Volume 284, Issue 44, Page(s) 30526–30533

    Abstract: Human interleukin-24 (IL-24) is unique among the IL-10 superfamily as there is considerable evidence that it possesses multiple anti-cancer properties, including direct tumor cell cytotoxicity, helper T cell (TH1) immune stimulation, and anti-angiogenic ... ...

    Abstract Human interleukin-24 (IL-24) is unique among the IL-10 superfamily as there is considerable evidence that it possesses multiple anti-cancer properties, including direct tumor cell cytotoxicity, helper T cell (TH1) immune stimulation, and anti-angiogenic activities. The primary sequence of human IL-24 differs from homologous cytokines, because it possesses three consensus N-linked glycosylation sites and the potential for a single disulfide bond. To address the significance of these modifications in human IL-24, we analyzed the relationship between post-translational modifications and the cytokine activity of the human IL-24 protein. In contrast to related interleukins, we identified a relationship between net glycosylation, protein solubility, and cytokine activity. In addition, abrogation of the two cysteine residues by mutagenesis dramatically altered the ability of IL-24 to secrete from host cells and resulted in the concomitant loss of IL-24 activity. We conclude that, unlike other IL-10 family members, human IL-24 must be glycosylated to maintain solubility and bioavailability. Further, a single, unique disulfide bond is required for secretion and activity. These structure-function relationships show that, although IL-24 is a member of the IL-19 subfamily of IL-10-like cytokines by sequence similarity, its surface properties and its distinctive disulfide arrangement make it unique. These observations could explain the novel biological activities measured of this cytokine. Understanding the structural basis of IL-24 activity will be important in the interpretation of the function of this cytokine and in the development of scale-up strategies for biophysical and clinical applications.
    MeSH term(s) Cysteine/genetics ; Cytokines ; Disulfides ; Glycosylation ; Humans ; Interleukins/biosynthesis ; Interleukins/chemistry ; Interleukins/immunology ; Interleukins/metabolism ; Protein Conformation ; Protein Processing, Post-Translational/physiology ; Solubility ; Structure-Activity Relationship
    Chemical Substances Cytokines ; Disulfides ; Interleukins ; interleukin-24 ; Cysteine (K848JZ4886)
    Language English
    Publishing date 2009-09-04
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1074/jbc.M109.036061
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article: Adenoviral endoplasmic reticulum-targeted mda-7/interleukin-24 vector enhances human cancer cell killing.

    Pataer, Abujiang / Hu, Wenxian / Xiaolin, Lu / Chada, Sunil / Roth, Jack A / Hunt, Kelly K / Swisher, Stephen G

    Molecular cancer therapeutics

    2008  Volume 7, Issue 8, Page(s) 2528–2535

    Abstract: We developed several adenoviral vectors designed to target MDA-7 expression to different subcellular compartments [endoplasmic reticulum (ER), mitochondria, nucleus, and cytosol] and evaluated their ability to enhance apoptosis. Adenoviral ER-targeted ... ...

    Abstract We developed several adenoviral vectors designed to target MDA-7 expression to different subcellular compartments [endoplasmic reticulum (ER), mitochondria, nucleus, and cytosol] and evaluated their ability to enhance apoptosis. Adenoviral ER-targeted mda-7/interleukin-24 vector (Ad-ER-mda7) selectively and effectively inhibited the growth and proliferation of lung (A549 and H1299) and esophageal (Seg1 and Bic1) cancer cells by enhancing cell killing. Both Ad-mda7 and Ad-ER-mda7 activated a novel pathway of ER stress-induced apoptosis characterized by unregulated expression of phosphorylated JNK, phosphorylated c-Jun, and phosphorylated RNA-dependent protein kinase. Caspase-4 activation mediated Ad-mda7- and Ad-ER-mda7-induced cell death. In addition, Ad-mda7- and Ad-ER-mda7-mediated growth inhibition correlated with activation of ER molecular markers RNA-dependent protein kinase and JNK both in vitro (in Ad-mda7- or Ad-ER-mda7-treated lung cancer cells) and in vivo. These findings suggest that vectors targeting the ER (Ad-ER-mda7) may be more effective in cancer gene therapy possibly through more effective induction or ER stress pathways.
    MeSH term(s) Adenoviridae/genetics ; Animals ; Cell Line, Tumor ; Endoplasmic Reticulum/metabolism ; Female ; Fluorescent Antibody Technique ; Genetic Vectors ; Interleukins/genetics ; Mice ; Mice, Nude ; Neoplasms/pathology
    Chemical Substances Interleukins ; interleukin-24
    Language English
    Publishing date 2008-08-21
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2063563-1
    ISSN 1538-8514 ; 1535-7163
    ISSN (online) 1538-8514
    ISSN 1535-7163
    DOI 10.1158/1535-7163.MCT-08-0083
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article: Structural Mapping of Post-translational Modifications in Human Interleukin-24: ROLE OF N-LINKED GLYCOSYLATION AND DISULFIDE BONDS IN SECRETION AND ACTIVITY

    Fuson, Kerry L / Zheng, Mingzhong / Craxton, Molly / Pataer, Abujiang / Ramesh, Rajagopal / Chada, Sunil / Sutton, R. Bryan

    Journal of biological chemistry. 2009 Oct. 30, v. 284, no. 44

    2009  

    Abstract: Human interleukin-24 (IL-24) is unique among the IL-10 superfamily as there is considerable evidence that it possesses multiple anti-cancer properties, including direct tumor cell cytotoxicity, helper T cell (TH1) immune stimulation, and anti-angiogenic ... ...

    Abstract Human interleukin-24 (IL-24) is unique among the IL-10 superfamily as there is considerable evidence that it possesses multiple anti-cancer properties, including direct tumor cell cytotoxicity, helper T cell (TH1) immune stimulation, and anti-angiogenic activities. The primary sequence of human IL-24 differs from homologous cytokines, because it possesses three consensus N-linked glycosylation sites and the potential for a single disulfide bond. To address the significance of these modifications in human IL-24, we analyzed the relationship between post-translational modifications and the cytokine activity of the human IL-24 protein. In contrast to related interleukins, we identified a relationship between net glycosylation, protein solubility, and cytokine activity. In addition, abrogation of the two cysteine residues by mutagenesis dramatically altered the ability of IL-24 to secrete from host cells and resulted in the concomitant loss of IL-24 activity. We conclude that, unlike other IL-10 family members, human IL-24 must be glycosylated to maintain solubility and bioavailability. Further, a single, unique disulfide bond is required for secretion and activity. These structure-function relationships show that, although IL-24 is a member of the IL-19 subfamily of IL-10-like cytokines by sequence similarity, its surface properties and its distinctive disulfide arrangement make it unique. These observations could explain the novel biological activities measured of this cytokine. Understanding the structural basis of IL-24 activity will be important in the interpretation of the function of this cytokine and in the development of scale-up strategies for biophysical and clinical applications.
    Language English
    Dates of publication 2009-1030
    Size p. 30526-30533.
    Publishing place American Society for Biochemistry and Molecular Biology
    Document type Article
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    Database NAL-Catalogue (AGRICOLA)

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  8. Article ; Online: Double-stranded RNA-dependent protein kinase-dependent apoptosis induction by a novel small compound.

    Hu, Wenxian / Hofstetter, Wayne / Wei, Xiaoli / Guo, Wei / Zhou, Yanbin / Pataer, Abujiang / Li, Hong / Fang, Bingliang / Swisher, Stephen G

    The Journal of pharmacology and experimental therapeutics

    2008  Volume 328, Issue 3, Page(s) 866–872

    Abstract: The interferon-induced, double-stranded RNA-dependent protein kinase (PKR) can play critical roles in inhibiting virus replication and inducing apoptosis. To develop new agents that may inhibit viral replication or induce apoptosis in cancer cells via ... ...

    Abstract The interferon-induced, double-stranded RNA-dependent protein kinase (PKR) can play critical roles in inhibiting virus replication and inducing apoptosis. To develop new agents that may inhibit viral replication or induce apoptosis in cancer cells via the PKR signaling pathway, we screened a chemical library for compounds that have differential cytotoxic effects on wild-type [mouse embryonic fibroblast (MEF)/PKR(+/+)] and PKR-knockout [MEF/PKR(-/-)] mouse embryonic fibroblast cells. We identified a synthetic compound, BEPP [1H-benzimidazole1-ethanol,2,3-dihydro-2-imino-a-(phenoxymethyl)-3-(phenylmethyl)-,monohydrochloride], that induces a cytotoxic effect more effectively in MEF/PKR(+/+) cells than in MEF/PKR(-/-) cells. BEPP also relatively effectively inhibited the growth of a human lung cancer cell line overexpressing PKR, compared with other cancer cell lines. In sensitive cells, BEPP induced apoptosis with activation of caspase-3. Treatment with BEPP led to increased phosphorylation of PKR and eIF2alpha, increased expression of BAX, and decreased expression of Bcl-2. BEPP-induced apoptosis was PKR dependent and was blocked by the adenovector expressing the dominant-negative PKR. Furthermore, pretreatment of HeLa cells at a noncytotoxic dose of BEPP effectively inhibited Vaccinia virus replication. Together, our results suggest that BEPP and its analogs may induce PKR-dependent apoptosis and inhibition of viral replication and that they can be a potential anticancer or anti-virus agent.
    MeSH term(s) Animals ; Apoptosis/drug effects ; Apoptosis/physiology ; Benzimidazoles/pharmacology ; Cell Line, Tumor ; Cell Survival/drug effects ; HeLa Cells ; Humans ; Lung Neoplasms ; Mice ; Mice, Knockout ; RNA, Double-Stranded/genetics ; Vaccinia virus/drug effects ; Virus Replication/drug effects ; eIF-2 Kinase/deficiency ; eIF-2 Kinase/genetics ; eIF-2 Kinase/metabolism
    Chemical Substances Benzimidazoles ; RNA, Double-Stranded ; eIF-2 Kinase (EC 2.7.11.1)
    Language English
    Publishing date 2008-12-09
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 3106-9
    ISSN 1521-0103 ; 0022-3565
    ISSN (online) 1521-0103
    ISSN 0022-3565
    DOI 10.1124/jpet.108.141754
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article: Adenoviral melanoma differentiation-associated gene 7 induces apoptosis in lung cancer cells through mitochondrial permeability transition-independent cytochrome c release.

    Pataer, Abujiang / Chada, Sunil / Hunt, Kelly K / Roth, Jack A / Swisher, Stephen G

    The Journal of thoracic and cardiovascular surgery

    2003  Volume 125, Issue 6, Page(s) 1328–1335

    Abstract: Objective: Melanoma differentiation-associated gene 7 is a novel tumor suppressor gene that induces apoptosis in lung cancer cells when delivered by adenoviral gene transfer as Ad-mda7. The mechanisms of action are not well defined but may involve ... ...

    Abstract Objective: Melanoma differentiation-associated gene 7 is a novel tumor suppressor gene that induces apoptosis in lung cancer cells when delivered by adenoviral gene transfer as Ad-mda7. The mechanisms of action are not well defined but may involve release of cytochrome c from the mitochondria with subsequent caspase activation.
    Methods: The lung cancer cell lines A549 and H1299 were transduced with Ad-mda7, adenovirus containing the gene for p53 (Ad-p53), and control adenoviral luciferase vectors. Staurosporine was used as a positive control to induce cytochrome c release through mitochondrial permeability transition-dependent pores, whereas cyclosporine (INN: ciclosporin) was used to specifically inhibit these mitochondrial permeability transition-dependent pores. Apoptosis was evaluated with fluorescence-activated cell sorting analysis of subdiploid populations and mitochondrial membrane potential changes with tetramethylrhodamine ethylester perchlorate.
    Results: Melanoma differentiation-associated gene 7, transduced by Ad-mda7 into H1299 and A549 lung cancer cells, resulted in sharp increases in cytosolic cytochrome c levels followed by induction of apoptosis and cellular death. The release of cytochrome c from the mitochondria occurred without changes in the mitochondrial membrane potential. Unlike staurosporine treatment, transduction with Ad-p53 and Ad-mda7 caused releases of cytochrome c and apoptosis that were not blocked by cyclosporine, suggesting a mitochondrial permeability transition pore-independent pathway.
    Conclusions: Ad-mda7 induces apoptosis in lung cancer cells through mitochondrial cytochrome c release in a process that is not dependent on mitochondrial membrane potential changes and occurs through mitochondrial permeability transition-independent pores. This unique mechanism of action may allow treatment of patients with lung cancer resistant to mitochondrial permeability transition-dependent cell death processes.
    MeSH term(s) Adenoviridae ; Apoptosis/drug effects ; Caspases/metabolism ; Cyclosporine/pharmacology ; Cytochrome c Group/metabolism ; Enzyme Activation ; Gene Transfer Techniques ; Genes, Tumor Suppressor/physiology ; Genes, p53/physiology ; Humans ; Interleukins/pharmacology ; Lung Neoplasms/pathology ; Membrane Potentials/physiology ; Mitochondria/enzymology ; Mitochondria/physiology ; Staurosporine/pharmacology ; Transduction, Genetic ; Tumor Cells, Cultured
    Chemical Substances Cytochrome c Group ; Interleukins ; interleukin-24 ; Cyclosporine (83HN0GTJ6D) ; Caspases (EC 3.4.22.-) ; Staurosporine (H88EPA0A3N)
    Language English
    Publishing date 2003-04-28
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 3104-5
    ISSN 1097-685X ; 0022-5223
    ISSN (online) 1097-685X
    ISSN 0022-5223
    DOI 10.1016/s0022-5223(02)73247-9
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Prognostic significance of RNA-dependent protein kinase on non-small cell lung cancer patients.

    Pataer, Abujiang / Raso, Maria Gabriela / Correa, Arlene M / Behrens, Carmen / Tsuta, Koji / Solis, Luisa / Fang, Bingliang / Roth, Jack A / Wistuba, Ignacio I / Swisher, Stephen G

    Clinical cancer research : an official journal of the American Association for Cancer Research

    2010  Volume 16, Issue 22, Page(s) 5522–5528

    Abstract: Purpose: The role of RNA-dependent protein kinase (PKR) in antiviral defense mechanisms and in cellular differentiation, growth, and apoptosis is well known, but the role of PKR in human lung cancer remains poorly understood. To explore the role of PKR ... ...

    Abstract Purpose: The role of RNA-dependent protein kinase (PKR) in antiviral defense mechanisms and in cellular differentiation, growth, and apoptosis is well known, but the role of PKR in human lung cancer remains poorly understood. To explore the role of PKR in human lung cancer, we evaluated the expression of PKR in tissue microarray (TMA) specimens from both non-small cell lung cancer (NSCLC) and normal human bronchial epithelium tissue.
    Experimental design: TMA samples (TMA-1) from 231 lung cancers were stained with PKR antibody and validated on TMA-2 from 224 lung cancers. Immunohistochemical expression score was quantified by three pathologists independently. Survival probability was computed by the Kaplan-Meier method.
    Results: The NSCLC cells showed lower levels of PKR expression than normal bronchial epithelium cells did. We also found a significant association between lower levels of PKR expression and lymph node metastasis. We found that loss of PKR expression is correlated with a more aggressive behavior, and that a high PKR expression predicts a subgroup of patients with a favorable outcome. Univariate and multivariate Cox proportional hazards regression models showed that a lower level of PKR expression was significantly associated with shorter survival in NSCLC patients. We further validated and confirmed PKR to be a powerful prognostic factor in TMA-2 lung cancer (hazard ratio, 0.22; P < 0.0001).
    Conclusions: Our findings first indicate that PKR expression is an independent prognostic variable in NSCLC patients.
    MeSH term(s) Bronchi/cytology ; Bronchi/enzymology ; Bronchi/metabolism ; Carcinoma, Non-Small-Cell Lung/diagnosis ; Carcinoma, Non-Small-Cell Lung/enzymology ; Carcinoma, Non-Small-Cell Lung/metabolism ; Female ; Humans ; Immunohistochemistry ; Kaplan-Meier Estimate ; Lung Neoplasms/diagnosis ; Lung Neoplasms/enzymology ; Lung Neoplasms/metabolism ; Male ; Prognosis ; Tissue Array Analysis ; eIF-2 Kinase/biosynthesis
    Chemical Substances eIF-2 Kinase (EC 2.7.11.1)
    Language English
    Publishing date 2010-10-07
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 1225457-5
    ISSN 1557-3265 ; 1078-0432
    ISSN (online) 1557-3265
    ISSN 1078-0432
    DOI 10.1158/1078-0432.CCR-10-0753
    Database MEDical Literature Analysis and Retrieval System OnLINE

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