ABSTRACT
OBJECTIVE: To explore a simple and practical method for human primary lung cancer cells culture in vitro. METHODS: Tumor specimens from 6 lung cancer patients were isolated with collagenase digestion cultured in vitro. Then the characteristics of these cells were analyzed and identified by optical microscope observation, hematoxylin-eosin staining, immunocytochemistry, immunohistochemistry and tumor nude mice inoculation experiments, respectively. RESULTS: Except for the small cell lung cancer, the other 5 samples were successfully isolated and cultured. The cultured cells showed typical characteristics of malignant cells and positive for cytokeratin 7 and 19. Moreover, the cancer cells readily formed subcutaneous tumors in nude mice and the pathological images of the transplanted tumor were consistent with its tumor origin. CONCLUSION: The primary culture for human lung cancer cells can be successfully achieved with the method of collagenase digestion.
Subject(s)
Adenocarcinoma/pathology , Carcinoma, Adenosquamous/pathology , Carcinoma, Squamous Cell/pathology , Cell Separation/methods , Lung Neoplasms/pathology , Primary Cell Culture , Small Cell Lung Carcinoma/pathology , Adenocarcinoma/metabolism , Aged , Animals , Biomarkers, Tumor/metabolism , Carcinoma, Adenosquamous/metabolism , Carcinoma, Squamous Cell/metabolism , Cell Proliferation , Collagenases/metabolism , Female , Humans , Keratin-19/metabolism , Keratin-7/metabolism , Lung Neoplasms/metabolism , Male , Mice, Inbred BALB C , Mice, Nude , Middle Aged , Neoplasm Transplantation , Small Cell Lung Carcinoma/metabolism , Time Factors , Tumor Burden , Tumor Cells, CulturedABSTRACT
Despite advances in early diagnosis and multimodality therapy for cancers, most of lung cancer patients have been locally advanced or metastatic at the time of diagnosis, suggesting the highly progressive characteristic of lung cancer cells. The mechanisms underling invasiveness and metastasis of lung cancer are yet to be elucidated. In the present study, immunohistochemistry was performed to detect the expression of CXCL16-CXCR6 in human lung cancer tissues. It was demonstrated that similar to CXCL12 and CXCR4, CXCL16 and CXCR6 were also coexpressed in human primary lung cancer tissues. After confirming the functional existence of CXCL16 and CXCR6 protein in A549, 95D and H292 cells by ELSA and flow cytometry analysis, we further explored the significance of CXCL16-CXCR6 axis in the biological functions of lung cancer cell lines in vitro. It was found that CXCL16 had no effects on the PCNA (proliferating cell nuclear antigen) expression of A549, 95D and H292 cells. However, both exogenous CXCL16 and CM (conditioned medium from A549, 95D or H292) significantly improved the in vitro viability and invasion of three lung cancer cell lines. The neutralizing antibody to CXCL16 or down-regulation of CXCR6 was able to inhibit the increased viability and invasiveness of A549, 95D and H292 cells stimulated by CXCL16 or CM. Our results imply that CXCL16-CXCR6 axis is involved in the regulation of viability and invasion rather than PCNA expression of lung caner cells, which opens the door for better understanding the mechanisms of lung tumor progression and metastasis.
