Effect of primarily cultured human lung cancer-associated fibroblasts on radiosensitivity of lung cancer cells / 中华放射肿瘤学杂志

Objective To investigate the effect of human lung cancer-associated fibroblasts (CAF) on the radiosensitivity of lung cancer cells when CAF is placed in direct contact co-culture with lung cancer cells.Methods Human lung CAF was obtained from fresh human lung adenocarcinoma tissue specimens by primary culture and subculture and was then identified by immunofluorescence staining.The CAF was placed in direct contact co-culture with lung cancer A549 and H1299 cells,and the effects of CAF on the radiosensitivity of A549 and H1299 cells were evaluated by colony-forming assay.Results The human lung CAF obtained by adherent culture could stably grow and proliferate,and it had specific expression of α-smooth muscle actin,vimentin,and fibroblast activation protein,but without expression of cytokeratin-18.The plating efficiency (PE,％) of A549 cells at 0 Gy irradiation was (20.0 ± 3.9) ％ when cultured alone versus (32.3 ± 5.5) ％ when co-cultured with CAF (t =3.16,P ＜ 0.05),and the PE of H1299 cells at 0 Gy irradiation was (20.6 ± 3.1) ％ when cultured alone versus (35.2 ± 2.3) ％ when co-cultured with CAF (t =6.55,P ＜0.05).The cell survival rate at 2 Gy irradiation (SF2) of A549 cells was 0.727 ±0.061 when cultured alone versus 0.782 ± 0.089 when co-cultured with CAF (t =0.88,P ＞ 0.05),and the SF2 of H1299 cells was 0.692 ±0.065 when cultured alone versus 0.782 ± 0.037 when co-cultured with CAF (t =2.08,P ＞0.05).The protection enhancement ratios of human lung CAF for A549 cells and H1299 cells were 1.29 and 1.25,respectively.Conclusions Human lung CAF reduces the radiosensitivity of lung cancer cells when placed in direct contact co-culture with them,and the radioprotective effect may be attributed to CAF promoting the proliferation of lung cancer cells.

Inhibitory Effect of Epidermal Growth Factor on the Proliferation of Lung Cancer Cell Lines

PURPOSE: Epidermal growth factor (EGF) is known to cause cellular proliferation, differentiation and survival. However, there are a few articles that have reported on the cell killing effect of EGF. We evaluated the effects of EGF on the survival of some lung cancer cell lines and we tried to determine the mechanism of action. MATERIALS AND METHODS: We examined various lung cancer cell lines. The cultured cells were exposed to radiation alone (0, 2, 5, and 10 Gy), EGF alone (0~1,000 nM) or a combination of radiation and EGF (10 nM). Survival was measured using a clonogenic assay and the expressions of epidermal growth factor receptor (EGFR), Ki-67 and cleaved-PARP were detected by western blot analysis. K-ras mutations were detected using polymerase chain reaction and sequencing. RESULTS: Treatment of EGF induced cell death in the lung cancer cell lines. The addition of EGF (10 nM) to radiation (0, 2, 5, and 10 Gy) resulted in an increased cell killing effect of radiation. The EGFR expression decreased with the addition of EGF. EGF increased the expression of cleaved-PARP, but it did not increase the expression of Ki-67. The effects of EGF were not correlated with EGFR mutation or K-ras mutation. CONCLUSION: In our study, EGF inhibited the proliferation of lung cancer cell lines and it induced apoptosis. EGF enhanced the radiosensitivity of lung cancer cells when EGF was combined with radiation. It is suggested that EGF seem to be one of the cytotoxic agents for lung cancer cell lines.

ASPS induces G_2/M arrest of H446 cells by activation of ERK signal pathway / 基础医学与临床

Objective To investigate ASPS induced G_2/M arrest in lung cancer cell line H446 and its effect on ERK MAP kinase signal transduction pathways. Methods Cell cycle phases were inspected by flow cytometery (FCM) ; Western blot analysis was used to inspect the proteins of ERK, p-ERK. Results Compared with control group, G_2/M phase cells increased with concentration significantly, G_0/G_1 phase cells were not different, G_2/M phase cells and G_0/G_1 phase cells were not different when pre-incubated with PD98059 prior to exposure to ASPS of different concentrations, protein of p-ERK was significantly increased, expression of ERK was no different. Conclusion ASPS may induce G_2/M arrest of H446 cells possibly by activation ERK MAP kinase pathways.

The primary study about the relationship of FBN2 gene and lung cancer cell lines / 重庆医科大学学报

Objective:To study the gene expression and aberrant methylation of FBN2 on lung cancer cell lines.Methods:Three lung cancer cell lines HCC366,H1299 and H2195 were cultured.mRNA was analysed by RT-PCR and methylation status was also investigated by methylation specific PCR.The loss of FBN2 expression cell lines were treated with the demethylating agent,5-aza-2'-deoxycytid-ine(5-aza-cdR).Results:The expression of FBN2 was detected in NHBEC and H2195,whereas it was not detected in HCC366 and H1299 which showed aberrant methylation of FBN2.FBN2 expression was restored after treatment with 5-aza-cdR.Conclusion:Methylation and silencing of FBN2 in tumor cells may play an important role in carcinogenesis of lung cancer.

The Effect of Glutathione on High Dose Cisplatin-Induced Cellular Toxicity in Non-small Cell Lung Cancer Cell Lines / 결핵

BACKGROUND: This study was designed to examine how glutathione, one of the nucleophilic sulfur compounds, effects the cisplatin cellular toxicity in the non-small cell lung cancer cell lines and normal lung epithel ial cell line. METHODS: Three cultured cell lines, the lung adenocarcinoma cell(NCL-H23), the lung squamous carcinoma cell (SK-MES-1) and the normal lung epithelial cell(L-132) line were exposed to various concentrations of cisplation with or without glutathione. The relative viability was estimated as a means of measuring the cisplatin cellular toxicity using the MTT method. RESULTS: In NCL-23, the response to cisplatin was sensitive but glutathione markedly increased the relative survival of the tumor cells by removing the antitumor effect of cisplatin. In both SK-MES-1 and L-132, the responses to cisplatin were less sensitive, and the chemoprotective effect of glutathione compared to an equal cisplatin dose was signigicantly higher in L-132 than in SK-MES-1(p<0.05). CONCLUSION: The protective effects of glutathione on cisplatin-induced cellular toxicity is more signigicant in normal lung epithelial cells than in squamous carcinoma cells.

Limited Cytotoxic Effect of Adenoviral-mediated p53 Gene Transfer in Variable Non-small Cell Lung Cancer (NSCLC) Cell Lines / 대한암학회지

PURPOSE: Cancer gene therapeutic strategy using p53 tumor suppresser gene have been suggested to be effective in many solid tumors including non-small cell lung cancer (NSCLC). To test generalized applicability, we tested a number of non-small cell lung cancer cell lines for their sensitivity to adenoviral-mediated wild-type p53 gene transfer. MATERIALS AND METHOD: Replication-incompetent recombinant adenovirus encoding wild- type p53 (Avp53) under the control of the human cytomegalovirus (CMV) promoter was constructed and the cytotoxic effectiveness was evaluated in various NSCLC cell lines. Because 20 moi (multiplicity of infection; number of active virus particle/cell number) of Avp53 showed highly-effective cytotoxicity in p53-deleted cell lines (NCI-H1299, and NCI-H358), same amount was used for other cell lines. RESULTS: Variable degree of cytotoxicity were observed in cell line with p53 mutation, but almost no effect were observed in those with will-type p53. Neither the infectivity of adenovirus, which was observed by x-gal stain after adenoviral mediated lac Z gene, nor the expression of p53 protein in infected cell, which was observed by western blot, was not the useful marker to expect the cytotoxic effect of Avp53. However, in responsive cell lines with Avp53, prominent expression of p21 protein, which was observed by western blot, was noticed. CONCLUSION: In conclusion, adenoviral-mediated wild-type p53 transfer may not be applicable to every patient with non-small cell lung cancer, especially when the tumor has wild-type p53 gene. Better method to predict the effectiveness before application and strategy to widen the applicable extent is needed.