RESUMEN
Purpose To investigate the protein and mRNA expression of c-IAPl in gastric adenocarcinoma tissue and its clinical significance. Methods Immunohistochemistry technique, Western blot and realtime fluorescent quantitative PCR(qRT-PCR) were used to detect the protein and mRNA expression of c-IAPl in 50 cases of gastric adenocarcinoma tissues and40 cases of adjacent normal gastric mucosa tissues to analyze its role in the development and progression of gastric adenocarcinoma, the relationship between the protein and mRNA expression of c-IAPl and the clinicopathological features. Results The relative expression level of c-IAPl protein in gastric adenocarcinoma tissues was significantly higher than that in adjacent normal gastric mucosa tissues, the difference was statistically significant (P＜ 0.05 ). The mRNA expression of c-IAPl in gastric adenocarcinoma was significantly higher than normal gastric mucosa tissues, the difference was statistically significant (P＜0.05). The protein and mRNA expression of c-IAPl were correlated with the degree of differentiation of gastric adenocarcinoma tissues, TNM clinical stage, lymph node metastasis and infiltration depth, the difference was statistically significant (P＜0.05 ), while there was no correlation with gender and age, the difference was not statistically significant (P＞ 0.05). Conclusion The high protein and mRNA expression of c-IAPl in gastric adenocarcinoma tissues inhibit the apoptosis of gastric adenocarcinoma cells, which contribute to the development and progression of gastric carcinoma and it may provide a new theoretical basis for the clinical targeted therapy of gastric adenocarcinoma.
RESUMEN
Objective To explore the effect of cellular inhibitor of apoptosis protein1 (cIAP-1) gene on the radiosensitivity of SMMC-7721 cells.Methods We silenced cIAP-1 expressions by the shRNA technology,and then we detected the changes of cell proliferation,cell cycle and cell apoptosis by CCK8 as-say,Western blot,qRT-PCR and flow cytometry after the radiotherapy.Results The cell proliferation rate of liver cancer SMMC-7721 cells at different radiation doses of 1 Gy,4 Gy,7 Gy and 10 Gy was detected.Comparing with control group (pGCsi-H1-control),the cell proliferation in cIAP-1 silencing group (pGCsiH1-shRNA) was significantly reduced at various radiation doses,and the effect was dose-dependent (P ＜ 0.05).G1/G0 phase arrest was observed after radiation (P ＜0.01),and the proportion of cells in S phase was significantly reduced compared with control (P ＜ 0.01).Compared with control group,G1/G0 phase arrest was detected (P ＜ 0.05),and the percentage of cell apoptosis was increased significantly in cIAP-1 silencing group (P ＜ 0.05).Conclusion cIAP-1 silencing can enhance the radiosensitivity of liver cancer cells,and inhibit cell proliferation by promoting the anti-tumor effect of radiation.
RESUMEN
[Abstratc] Objective The function of cIAP1 in the progression of ovarian cancer has not been clarified . This study is to explore the involvement of cIAP 1 in regulating biological behaviors of ovarian cancer cells by u-sing RNA interference(RNAi)technology.Mte hods The short hairpin RNA plasmid targeting cIAP1 was con-structed and transfected into Skov 3 cells.The levels of cIAP1 mRNA and protein were investigated by RT -PCR and Western Blot respectively .MTT assay and flow cytometry were used to evaluate cell proliferation and apopto-sis.R esults The rate of cIAP1 transfection was 74.7%performed by flow cytometric analysis .cIAP1 expression was significantly down -regulated at both mRNA and protein levels ,which resulted in a decrease of cell prolifera-tion and invasion capability in vitro .Conclusion This study implies that cIAP 1 might play an important role in the progression of ovarian cancer ,and it could be a potential target for therapeutic anti -cancer drugs .
RESUMEN
BACKGROUND: Activation of the transcription factor NF-kappaB has been shown to protect cells from tumor necrosis factor-alpha, chemotherapy, and radiation-induced apoptosis. NF-kappaB-dependent cIAP expression is a major antiapoptotic mechanism for that. NF-kappaB activation and cIAP expression in A549 lung cancer cells which is relatively resistant to radiation-induced cell death were investigated for the mechanism of radioresistance. MATERIALS AND METHODS: We used A549 lung cancer cells and Clinac 1800C linear accelerator for radiation. Cell viability test was done by MTT assay. NF-kappaB activation was tested by luciferase reporter gene assay, Western blot for IkappaBalpha degradation, and electromobility shift assay. For blocking NF-kappaB, MG132 and transfection of IkappaBalpha-superrepressor plasmid construct were used. cIAP expression was analyzed by RT-PCR and cIAP2 promoter activity was performed using luciferase assay system. RESULTS: MTT assay showed that cytotoxicity even 48 hr after radiation in A549 cells were less than 20%. Luciferas assay demonstrated weak NF-kappaB activation of 1.6+/-0.2 fold compared to PMA-induced 3.4+/-0.9 fold. Radiation-induced IkappaBalpha degradation was observed in Western blot and NF-kappaB DNA binding was confirmed by EMSA. However, blocking NF-kappaB using MG132 and IkappaBalpha-superrepressor transfection did not show any sensitizing effect for radiation-induced cell death. The result of RT-PCR for cIAP1 & 2 expression was negative induction while TNF-alpha showed strong expression for cIAP1 & 2. The cIAP2 promoter activity also did not show any change compared to positive control with TNF-alpha. CONCLUSIONS: We conclude that activation of NF-kappaB does not determine the intrinsic radiosensitivity of cancer cells, at least for the cell lines tested in this study.