ABSTRACT
An experimental model, in which exogenous gene expression in the lung is achieved, has been established. A fibroblast cell line was transfected with the lacZ gene and was administered to syngeneic mice by either intravenous or intratracheal injection. Enzyme-linked immunosorbent assay and histochemical detection of beta-galactosidase revealed efficient gene delivery to the lung by either route. Kinetic studies demonstrated that the expression peaked immediately after the injection, and this high level was maintained for 7 days by intratracheal and for 14 days by intravenous administration. This system may have potential relevance for certain experimental models requiring specific gene delivery to the lung.
Subject(s)
Fibroblasts/enzymology , Galactosidases/genetics , Gene Transfer Techniques , Gene Transfer, Horizontal , Lung/enzymology , Animals , Cell Transplantation , Female , Fibroblasts/cytology , Fibroblasts/transplantation , Galactosidases/metabolism , Genetic Vectors , Lac Operon/genetics , Lung/cytology , Mice , Mice, Inbred BALB C , Models, Animal , Specific Pathogen-Free Organisms , TransfectionABSTRACT
Endobronchial ultrasonography (US) with 4.5-F small-caliber US probes, combined with bronchoalveolar lavage technique, was evaluated in autopsied lungs and 22 patients with various pulmonary interstitial or alveolar diseases. Several different echoic patterns were found that may reflect changes due to pathologic alteration of lung parenchyma. This technique may have potential for evaluation and diagnosis of peripheral lung diseases.
Subject(s)
Endosonography , Lung Diseases/diagnostic imaging , Lung/diagnostic imaging , Adult , Aged , Bronchoalveolar Lavage Fluid , Female , Humans , Lung/pathology , Lung Diseases/pathology , Male , Middle Aged , Phantoms, ImagingABSTRACT
Ku70 protein, cooperating with Ku80 and DNA-dependent protein kinase (DNA-PK) catalytic subunit (DNA-PKcs), is involved in DNA double-strand break (DNA DSB) repair and V(D)J recombination. Recent studies have revealed increased ionizing radiosensitivity in Ku70-deficient cells. The presented study, using a human squamous cell lung carcinoma cell line, demonstrated that introduction of an antisense Ku70 nucleic acid made the cells more radio- and chemosensitive than the parental cells. Ku70 protein expression was suppressed in the cells with antisense Ku70 construct when compared to the wild-type cells. A relatively small but statistically significant increase in radiosensitivity of the cells was achieved by the introduction of the antisense Ku70. The increased radiosensitivity in vitro was accompanied by an approximately two-fold increase in alpha and alpha/beta values in a linear-quadratic model. The antisense Ku70 increased the chemosensitivity of the cells to some DNA-damaging agents such as bleomycin and methyl methanesulfonate, but not to cisplatin, mitomycin C, and paclitaxel. This system provides us with partial suppression of Ku70, and will be a useful experimental model for investigating the physiological roles of the DNA DSB repair gene.
Subject(s)
Carcinoma, Squamous Cell/drug therapy , Carcinoma, Squamous Cell/radiotherapy , DNA Helicases , DNA Repair/drug effects , DNA-Binding Proteins/antagonists & inhibitors , Lung Neoplasms/drug therapy , Lung Neoplasms/radiotherapy , Antigens, Nuclear/genetics , Antineoplastic Agents/therapeutic use , Blotting, Northern , Carcinoma, Squamous Cell/genetics , Colony-Forming Units Assay , DNA Damage , DNA Primers , DNA Repair/genetics , DNA, Antisense/pharmacology , DNA, Neoplasm/drug effects , DNA-Binding Proteins/genetics , Electrophoretic Mobility Shift Assay , Humans , In Vitro Techniques , Ku Autoantigen , Lung Neoplasms/genetics , Transfection , Tumor Cells, Cultured/drug effects , Tumor Cells, Cultured/radiation effects , X-RaysABSTRACT
Objective:To study the expression of the antisen se Ku70 in a human's lung carcinoma cell line(LCA) and its high-radiosensitivit y.Methods:The expression of the Ku70 protein in the Ku70- LCA and its radiosensitivity was detected by Western-blott ing and Cell Radiosensitivity Taste (to see The Cell Clonogenic Assay and Surviv al Curves).Results:(1)Only the sense Ku70 protein was defe cted in the Ku70-LCA,but the sense Ku70 protein wasn't defected in the Ku70as -LCA by Western-blotting.(2)The radiosensitivity was increased in the Ku70as- LCA and their radiosensitivity depend on the radiating dose(P＜0.01).Conclusion:After the expression of sense Ku70 protein in t he Ku70as-LCA was blocked by antisense Ku70,the Ku70as-LCA should be high-rad iosensitivit y,the Ku70 is a candiate target for cancer gene therapy.It was the first step fo r the study of tumor's specific radiating and gene therapy in the future.
