Associations between Comorbidities and Acute Exacerbation of Interstitial Lung Disease after Primary Lung Cancer Surgery

Acute exacerbation (AE) of interstitial lung disease (ILD) is a severe complication of lung resection in lung cancer patients with ILD (LC-ILD). This study aimed to assess the predictive value of comorbidities other than ILD for postoperative AE in patients with LC-ILD. We retrospectively evaluated 68 patients with LC-ILD who had undergone lung resection. We classified them into two groups: those who had developed postoperative AE within 30 days after resection and those who had not. We analyzed patient characteristics, high-resolution computed tomography findings, clinical data, pulmonary function, and intraoperative data. The incidence of postoperative AEs was 11.8%. In univariate analysis, performance status (PS), honeycombing, forced vital capacity (FVC), and high hemoglobin A1c (HbA1c) levels without comorbidities were significantly associated with postoperative AE. Patients were divided into two groups according to cutoff levels of those four variables as determined by receiver operating characteristic curves, revealing that the rates of patients without postoperative AE differed significantly between groups. The present results suggested that preoperative comorbidities other than ILD were not risk factors for postoperative AE in patients with LC-ILD. However, a high preoperative HbA1c level, poor PS, low FVC, and honeycombing may be associated with postoperative AE of LC-ILD

Cell Stress Induces Upregulation of Osteopontin via the ERK Pathway in Type II Alveolar Epithelial Cells

<div><p>Osteopontin (OPN) is a multifunctional protein that plays important roles in cell growth, differentiation, migration and tissue fibrosis. In human idiopathic pulmonary fibrosis and murine bleomycin-induced lung fibrosis, OPN is upregulated in type II alveolar epithelial cells (AEC II). However, the mechanism of OPN induction in AEC II is not fully understood. In this study, we demonstrate the molecular mechanism of OPN induction in AEC II and elucidate the functions of OPN in AEC II and lung fibroblasts. Human lung adenocarcinoma cells (A549) and mouse alveolar epithelial cells (MLE12), used as type II alveolar epithelial cell lines for in vitro assays, and human pulmonary alveolar epithelial cells (HPAEpiC) were treated with either bleomycin, doxorubicin or tunicamycin. The mechanism of OPN induction in these cells and its function as a pro-fibrotic cytokine on A549 and lung fibroblasts were analyzed. The DNA damaging reagents bleomycin and doxorubicin were found to induce OPN expression in A549, MLE12 and HPAEpiC. OPN expression was induced via activation of the extracellular signal-regulated protein kinase (ERK)-dependent signaling pathway in A549 and MLE12. The endoplasmic reticulum (ER) stress-inducing reagent tunicamycin induced <i>OPN</i> mRNA expression in A549, MLE12 and HPAEpiC, and <i>OPN</i> mRNA expression was induced via activation of the ERK-dependent signaling pathway in A549 and MLE12. Another ER stress-inducing reagent thapsigargin induced the expression of <i>OPN</i> mRNA as well as the subsequent production of OPN in A549 and MLE12. Furthermore, OPN promoted the proliferation of A549 and the migration of normal human lung fibroblasts. Inhibition of OPN by small interference RNA or neutralizing antibody suppressed both of these responses. The results of this study suggest that cell stress induces the upregulation of OPN in AEC II by signaling through the ERK pathway, and that upregulated OPN may play a role in fibrogenesis of the lung.</p></div

Radiation-quality-dependence in bystander cellular effects on normal human cells induced by carbon-ion and X-ray microbeams.

炭素イオン及びＸ線マイクロビームを低フルエンスでヒト正常細胞に照射した時に誘導される細胞致死と突然変異誘発に関する細胞応答（バイスタンダー効果）の線質依存性を調べた。マイクロビームを全細胞数の0.20%の細胞に限定的に照射した時、炭素イオンでは直接照射された細胞数を遙かに超えた細胞致死と突然変異誘発が観察された一方で、Ｘ線ではその様な現象は観察されなかった

Effect of bleomycin on OPN expression in type II alveolar epithelial cell lines.

<p>(A) A549 were exposed to 1 µg/mL bleomycin for 0, 6, 12, 24 and 48 h. <i>OPN</i> mRNA was assessed by qRT-PCR. (B) MLE12 were exposed to 1 µg/mL bleomycin for 0, 6, 12, 24 and 48 h. <i>Opn</i> mRNA was assessed by qRT-PCR. (C) A549 were exposed to 0, 1 and 10 µg/mL bleomycin for 48 h. <i>OPN</i> mRNA was assessed by qRT-PCR. (D) MLE12 were exposed to 0, 1 and 10 µg/mL bleomycin for 48 h. <i>Opn</i> mRNA was assessed by qRT-PCR. (E, F and G) A549 were exposed to 1 µg/mL bleomycin. (E) The cells were lysed at 0, 2, 4 and 6 d. OPN concentrations in cell lysates were determined by ELISA. (F) The cell culture supernatants were collected at 0, 2, 4 and 6 d. OPN concentrations in cell culture media were determined by ELISA. (G) Total OPN concentrations in cell lysates and cell culture media were determined by ELISA. Data are presented as mean ± S.E. *, P<0.05; **, P<0.01.</p

Effect of doxorubicin on OPN expression and molecular mechanism of OPN induction.

<p>(A and B) A549 were exposed to 50 nM doxorubicin or PBS for 48 h. (A) <i>OPN</i> mRNA was assessed by qRT-PCR. (B) OPN concentrations in cell culture media were determined by ELISA. (C) A549 were exposed to 50 nM doxorubicin or PBS for 24 h. Western blotting was used to detect phospho-specific ERK1/2 and ERK1/2. (D) A549 were pretreated with PD98059 (50 µM) or DMSO for 1 h, and subsequently exposed to 50 nM doxorubicin for 48 h. Western blotting was used to detect phospho-specific ERK1/2 and ERK1/2. (E) A549 were pretreated with U0126 (50 µM) or DMSO for 1 h, and subsequently exposed to 50 nM doxorubicin for 48 h. Western blotting was used to detect phospho-specific ERK1/2 and ERK1/2. (F) A549 were pretreated with PD98059 (50 µM) or DMSO for 1 h, and subsequently exposed to 50 nM doxorubicin or PBS for 48 h. <i>OPN</i> mRNA was assessed by qRT-PCR. (G) A549 were pretreated with U0126 (50 µM) or DMSO for 1 h, and subsequently exposed to 50 nM doxorubicin or PBS for 48 h. <i>OPN</i> mRNA was assessed by qRT-PCR. (H) A549 were pretreated with PD98059 (50 µM) or DMSO for 1 h, and subsequently exposed to 50 nM doxorubicin or PBS for 48 h. OPN concentrations in cell culture media were determined by ELISA. (I) A549 were pretreated with U0126 (50 µM) or DMSO for 1 h, and subsequently exposed to 50 nM doxorubicin or PBS for 48 h. OPN concentrations in cell culture media were determined by ELISA. Data are presented as mean ± S.E. *, P<0.05; **, P<0.01.</p

Effect of OPN on the proliferation of A549.

<p>(A and B) A549 were transfected with 20 nM human <i>OPN</i> siRNA or control siRNA. Twenty-four h after transfection, A549 were incubated for 48 h in the presence of 1 µg/mL bleomycin or PBS. (A) <i>OPN</i> mRNA was assessed by qRT-PCR. (B) OPN concentrations in cell culture media were determined by ELISA. (C and D) A549 were transfected with 20 nM human <i>OPN</i> siRNA or control siRNA. Twenty-four h after transfection, A549 were exposed to 1 µg/mL bleomycin for 48 h and then incubated with EdU for an additional 15 h. A549 were stained with Alexa Fluor 488 azide and HCS NuclearMask Blue stain. The cells were visualized by pseudocoloring with the Floid Cell Imaging Station. (C) Nuclei (i, ii) show representative photomicrographs of all cells. EdU (iii, iv) show representative photomicrographs of proliferating cells. Merge (v, vi) show representative photomicrographs of merged images. Bar: 100 µm. (D) Twelve fields per well were recorded and the number of positively-staining cells was counted. The percentages of proliferating A549 that were transfected with either human <i>OPN</i> siRNA or control siRNA are shown. Data are presented as mean ± S.E. *, P<0.05; **, P<0.01.</p

Effect of thapsigargin on OPN expression.

<p>(A) A549 were exposed to 10 nM thapsigargin or DMSO for 48 h. Western blotting was used to detect BiP and β-actin. (B and C) A549 were exposed to 10 nM thapsigargin or DMSO for 48 h. (B) <i>OPN</i> mRNA was assessed by qRT-PCR. (C) OPN concentrations in cell culture media were determined by ELISA. Data are presented as mean ± S.E. *, P<0.05; **, P<0.01.</p