// Roberto Gomez-Casal 1, 2 , Michael W. Epperly 1, 3 , Hong Wang 1, 4 , David A. Proia 5 , Joel S. Greenberger 1, 3 , Vera Levina 1, 2, 6 1 University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA 2 Department of Medicine, University of Pittsburgh, Pittsburgh, PA, USA 3 Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, PA, USA 4 Department of Biostatistics, University of Pittsburgh, Pittsburgh, PA, USA 5 Synta Pharmaceuticals Corp., Lexington, MA, USA 6 Current address: Hillman Cancer Center, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA Correspondence to: Vera Levina, e-mail: levinav@upmc.edu Keywords: lung adenocarcinoma, fractionated ionizing radiation, radioresistance mechanisms, DNA repair, heat shock protein-90 Received: August 14, 2015 Accepted: October 14, 2015 Published: October 27, 2015 ABSTRACT Despite the common usage of radiotherapy for the treatment of NSCLC, outcomes for these cancers when treated with ionizing radiation (IR) are still unsatisfactory. A better understanding of the mechanisms underlying resistance to IR is needed to design approaches to eliminate the radioresistant cells and prevent tumor recurrence and metastases. Using multiple fractions of IR we generated radioresistant cells from T2821 and T2851 human lung adenocarcinoma cells. The radioresistant phenotypes present in T2821/R and T2851/R cells include multiple changes in DNA repair genes and proteins expression, upregulation of EMT markers, alterations of cell cycle distribution, upregulation of PI3K/AKT signaling and elevated production of growth factors, cytokines, important for lung cancer progression, such as IL-6, PDGFB and SDF-1 (CXCL12). In addition to being radioresistant these cells were also found to be resistant to cisplatin. HSP90 is a molecular chaperone involved in stabilization and function of multiple client proteins implicated in NSCLC cell survival and radioresistance. We examined the effect of ganetespib, a novel HSP90 inhibitor, on T2821/R and T2851/R cell survival, migration and radioresistance. Our data indicates that ganetespib has cytotoxic activity against parental T2821 and T2851 cells and radioresistant T2821/R and T2851/R lung tumor cells. Ganetespib does not affect proliferation of normal human lung fibroblasts. Combining IR with ganetespib completely abrogates clonogenic survival of radioresistant cells. Our data show that HSP90 inhibition can potentiate the effect of radiotherapy and eliminate radioresistant and cisplatin -resistant residual cells, thus it may aid in reducing NSCLC tumor recurrence after fractionated radiotherapy.
Background/aim To determine whether Gramicidin S (GS)-nitroxide, JP4-039, esophageal radiation protection protected lung tumors in a transgenic model, LoxP-Stoop-LoxP Kristen Rat Sarcoma Viral oncogene (LSL-K-RAS) mice were administered intra-tracheal- Carbapenem-resistant Enterobacteriaceae (CRE) recombinase, bilateral lung tumors were confirmed at 11 weeks, then thoracic irradiation was delivered. Materials and methods Mice received single-fraction 15 Gy or 24 Gy to both lungs, in subgroups receiving intraesophageal administration 10 min before irradiation of JP4-039 (in F15 emulsion) tumor size reduction and survival were investigated. Mice were followed for survival, and reduction in tumor size. Results There was no evidence of tumor radioprotection in mice receiving JP4-039/F15. Conclusion Intraesophageal radioprotective small-molecule antioxidant therapy protects normal tissue but not tumor tissue in mice with transgenic lung tumors.
Aim:To explore the diagnostic value of serum cystatin C(Cys C) and lipoprotein(a) for early diabetic nephropathy(DN).Methods:The immune turbidimetry was used to detect serum Cys C and lipoprotein(a) in 60 cases of patients with early DN(DN group),55 patients without nephropathy but with diabetes mellitus(NDN group),and 50 healthy adults(control group).The efficacy of single index or 2 indexes combined in diagnosis of early DN was compared.Results:Serum Cys C and lipoprotein(a) in early DN group were significantly higher than those in the control group and NDN group(F=5.888,14.026,P0.05).The sensitivity of Cys C and lipoprotein(a) in diagnosis of DN was 86.7% and 68.3%,specificity was 95.0% and 93.3%,which were 91.6% and 91.7% when the 2 indexes combined.Conclusion:Combined detecting serum Cys C and lipoprotein(a) can greatly improve detection sensitivity of early DN.
Purpose: Patients with American Joint Committee on Cancer (AJCC) stage III melanoma are at high risk of recurrence and death. We hypothesized that a multiple-marker reverse transcriptase polymerase chain reaction (MM-RT-PCR) blood assay could predict, early in the course of therapy, those patients destined to experience treatment failure with a melanoma vaccine (MV) previously shown to improve survival in a phase II clinical trial. Patients and Methods: After complete surgical resection, prospectively collected cryopreserved peripheral-blood lymphocyte specimens (n = 90) from the serial bleeds of 30 patients with AJCC stage III melanoma were studied by MM-RT-PCR, using the markers tyrosinase, melanoma antigen recognized by T cells-1 (MART-1), and universal melanoma antigen gene-A (uMAG-A). All patients were enrolled in a phase II MV trial during the period of blood draws, and were selected for this study in a blinded fashion. Median duration of clinical follow-up was 74 months for the 13 survivors and 11 months for the 17 nonsurvivors. Results: The presence of at least one melanoma-specific RT-PCR marker was associated with an increased risk of disease recurrence (risk rate, 3.12; P = .02) and decreased risk of survival (relative rate, 2.62; P = .0496) by multivariate analysis. Conclusion: MM-RT-PCR of the blood provided early prediction of subsequent disease recurrence and death in clinically disease-free AJCC stage III melanoma patients enrolled in a MV phase II trial. On the basis of the results of this pilot study, the MM-RT-PCR blood assay should be considered as a clinically important monitoring tool for assessing patient response to adjuvant therapy, and in the surveillance of clinically disease-free patients for the earliest signs of recurrence.
<p>PDF file - 76K, Ex vivo frequencies of cytokine-producing NY-ESO-1 157-165-specific CD8+ T cells before and after vaccination with either MHC class I peptide alone (arm 1), or both MHC class I and class II peptides (arm 2).</p>
Objective To investigate expression of p16 gene in carcinoma of bladder and cystitis glandularis. Methods p16 gene expressions were analyzed in 49 patients with carcinoma of bladder and 10 patients with cystitis glandularis by immunohistochemical method. Results Positive expression rate of p16 protein in 49 patients with carcinoma of bladder was 38.78%. Positive expression had no relationship with histological type, differential degree, recurrence of carcinoma of bladder, and was correlative with infiltrative degree. The positive rate was 53.57% in not infiltrative,19.05% in infiltrative (P0.05) and p16 protein weren't expressed in 10 patients with cystitis glandularis. Conclusion Detection of p16 gene may be helpful in understanding biological features of carcinoma of bladder.
Host-cell cysteine proteases play an essential role in the processing of the viral spike protein of SARS coronaviruses. K777, an irreversible, covalent inactivator of cysteine proteases that has recently completed phase 1 clinical trials, reduced SARS-CoV-2 viral infectivity in several host cells: Vero E6 (EC50< 74 nM), HeLa/ACE2 (4 nM), Caco-2 (EC90 = 4.3 μM), and A549/ACE2 (<80 nM). Infectivity of Calu-3 cells depended on the cell line assayed. If Calu-3/2B4 was used, EC50 was 7 nM, but in the ATCC Calu-3 cell line without ACE2 enrichment, EC50 was >10 μM. There was no toxicity to any of the host cell lines at 10–100 μM K777 concentration. Kinetic analysis confirmed that K777 was a potent inhibitor of human cathepsin L, whereas no inhibition of the SARS-CoV-2 cysteine proteases (papain-like and 3CL-like protease) was observed. Treatment of Vero E6 cells with a propargyl derivative of K777 as an activity-based probe identified human cathepsin B and cathepsin L as the intracellular targets of this molecule in both infected and uninfected Vero E6 cells. However, cleavage of the SARS-CoV-2 spike protein was only carried out by cathepsin L. This cleavage was blocked by K777 and occurred in the S1 domain of the SARS-CoV-2 spike protein, a different site from that previously observed for the SARS-CoV-1 spike protein. These data support the hypothesis that the antiviral activity of K777 is mediated through inhibition of the activity of host cathepsin L and subsequent loss of cathepsin L-mediated viral spike protein processing.
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