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Objective: To investigate the effect of berbamine (BBM) on the induction of apoptosis in human lung cancer cell A549 cells and the activity of TNF-α-JNK signaling pathway. Methods: After the A549 cells being treated with BBM at different concentration, the inhibitory effect of BBM on proliferation was detected by MTT assay. Cell morphological changes were detected by light microscope. Alteration of apoptosis rate of A549 cells was determined by Annexin V/PI double staining. Western blotting was used to detect the activity of apoptosis-related proteins, including Bcl-x, Caspase-3, and PAPR. The expressions of c-jun N-terminal kinase (JNK) and p-JNK were determined by Western blotting. Changes of tumor necrosis factor-alpha (TNF-α) were detected by fluorescent quantitative PCR and ELISA, respectively. Finally, the impacts of BBM on the proliferation and apoptosis of A549 cells were detected in the absence or presence of a JNK inhibitor. Results: BBM significantly inhibited the growth of A549 cells in a dose-dependent manner. The IC50 of 24 h was 9.01 μmol/L. Cells treated with BBM showed the typically morphological characteristics of apoptotic cells. Annexin V/PI double staining test indicated that BBM could induce the apoptosis of A549 cells in a dose-dependent manner. The early apoptotic population of cells treated with 10 μmol/L BBM was 13.8%, which was 5.6 times higher than that of the control. BBM decreased the expression of anti-apoptotic protein Bcl-x and increased the activity of proapoptotic proteins of caspase-3 and PARP. The mRNA and the protein expression levels of TNF-α were significantly increased by BBM treatment. The p-JNK expression was also dramatically up-regulated after BBM treatment. The effects of BBM on the proliferation and apoptosis in A549 cells were significantly reduced when JNK pathway was blocked. Conclusion: BBM could inhibit the growth and induce the apoptosis of A549 cells, and its mechanism may be related to the activated TNF-α-JNK signaling pathway.
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Objective To investigate the effects of 15-deoxy-?12,14-prostaglandin J2(15d-PGJ2) and DDP on the growth of human pulmonary carcinoma PLA-801D cells and the mechanisms of apoptosis.Methods The human pulmonary carcinoma PLA-801D cells were selected and added to each well of 96-well place and cultivated for 24 h.Then the cells were treated with different concentrations of 15d-PGJ2(0,5,10,20,40 and 80 ?g?L-1) or 15d-PGJ2 combined with DDP(3 mg?L) for 24 h.0 ?g?L-1 15d-PGJ2 group was control group.The morphological changes of cells were observed under inverted microscope.Microculture tetrazolium(MTT)dye was applied to detect the proliferation of the human pulmonary carcinoma PLA-801D cells treated with 15d-PGJ2 and DDP.Diphenylamine assay(DPA) was used to evaluate the activation.Flow cytometry assay(FCM) was used to detect the apoptosis proportion and the changes of cell cycle.Results When the human pulmonary carcinoma PLA-801D cells were treated with low-concentration 15d-PGJ2 alone(5,10 and 20 ?g?L-1),no significant difference was observed in the inhibitory rate of cell growth and the apoptotic indexes such as the apoptosis proportion,the percent of DNA fragmentation and the activity of caspase-3 compared with control group(P
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Objective To investigate the antitumor effects of total glycoside of Cimicifuga dahurica (TGCD) in vivo and in vitro, and further explore its mechanisms. Methods The anti-tumor activity in vitro was determined by MTT assay and the anti-tumor activity in vivo was evaluated using experimental mouse tumor (S180) model and human tumor (A549) xenografts in nude mice. After treatment, A549 cell apoptosis and morphologic change were evaluated by Annexin V/PI flow cytometry and HE staining. Results Inhibitory concentration 50% (IC50) of TGCD on A549, HepG2, HL60, Eca-109 and MDA-MB231 cells were 20.3, 27.1, 21.2, 23.4 and 32.7 ?g/mL respectively. Administration of TGCD (100 or 200 mg/kg) inhibited S180 solid tumor development in mice, the inhibition rates were 42.8% and 54.6% respectively. Administration of TGCD (100 or 200 mg/kg) inhibited A549 tumor growth with a T/C (mean value of treated group/mean value of control group) value of 58.1% and 52.2% respectively. In addition, increased percentage of apoptotic cells induced by TGCD in human A549 nude mice xenografts and the histopathological changes including cell shrinkage and condensation of chromosomes were observed. Conclusion TGCD has demonstrated antitumor bioactivity both in vitro and in vivo, which may be related to its effects of inducing apoptosis activity.
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Three monoclonal antibodies against human pulmonory carcinoma, 4E_(12), 4F_(12)and 4H_2, were established with Xuan Wei human pulmonary adenocarcinoma cell line SLC-89 antigen. They had lasted for more than 8 months in Vitro. Having been stored in liquid nitrogen, the cells reviver normally and continued to secrete antibodies. The secreted antibodies could respond well to pulmonary carcinoma. Associated antigens in paraffin section were still stable. Therefore they will be valuable diagnostic reagents in histopathology.