ABSTRACT
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.
ABSTRACT
Objective To investigate the correlation between the level of apoptosis protein 1 (c-IAP1) and the radiosensitivity of lung cancer cells.Method The survival rate and proliferation of the lung cancer cells lines (A549,H460,H1299,H358,HCC827,H1650) from six human were detected by thiazolyl blue tetrazolium bromide (MTT) and cell colony formation assay.The DNA damage effects of radiation on lung cancer cells were detected by comet assay.The expressions of c-IAP1 protein and its mRNA were determined by Western blot and real-time quantitative PCR.Results The results of MTT and colony formation showed that the radiosensitivity of different lung cancer cells was also different,among which H358 and H460 cells had the highest radiosensitivity than that of H1650 and HCC827 cells,and H1299 and A549 cells had the weakest radiosensitivity.The results of comet assay showed that six kinds of lung cancer cells were suffered by DNA damage after radiation,and the DNA damage of H358 cells was most serious.The results of Western blot and real-time quantitative PCR showed that the c-IAP1 protein level was negatively correlated with the radiosensitivity of lung cancer cells.The higher the c-IAP1 protein level,the weaker the radiosensitivity of cells.The radiosensitivity was also affected by Smac protein levels.Conclusions c-IAP1 may be a selective target gene in mediating the radiosensitivity of lung cancer cells and this paper may contribute to the study of radioresistance and radiosensitization of cancer cell.
ABSTRACT
Objective To investigate the correlation between the level of apoptosis protein 1 (c-IAP1) and the radiosensitivity of lung cancer cells.Method The survival rate and proliferation of the lung cancer cells lines (A549,H460,H1299,H358,HCC827,H1650) from six human were detected by thiazolyl blue tetrazolium bromide (MTT) and cell colony formation assay.The DNA damage effects of radiation on lung cancer cells were detected by comet assay.The expressions of c-IAP1 protein and its mRNA were determined by Western blot and real-time quantitative PCR.Results The results of MTT and colony formation showed that the radiosensitivity of different lung cancer cells was also different,among which H358 and H460 cells had the highest radiosensitivity than that of H1650 and HCC827 cells,and H1299 and A549 cells had the weakest radiosensitivity.The results of comet assay showed that six kinds of lung cancer cells were suffered by DNA damage after radiation,and the DNA damage of H358 cells was most serious.The results of Western blot and real-time quantitative PCR showed that the c-IAP1 protein level was negatively correlated with the radiosensitivity of lung cancer cells.The higher the c-IAP1 protein level,the weaker the radiosensitivity of cells.The radiosensitivity was also affected by Smac protein levels.Conclusions c-IAP1 may be a selective target gene in mediating the radiosensitivity of lung cancer cells and this paper may contribute to the study of radioresistance and radiosensitization of cancer cell.
ABSTRACT
[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 .