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Objective: (1) observation of colon cancer cell lines SW480, HT29, HCT116 different sensitivity and three colon cancer cells c-Flip protein base expression levels of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). (2) to explore the role of the PI3K/Akt signaling pathway in TRAIL-induced colon cancer cells in the process of expression of c-Flip to reveal the mechanisms of colon cancer cells to TRAIL sensitivity change part. The Methods: colon cancer cell lines SW480, HT29, HCT116 study. (1) using the MTT assay and flow cytometry to detect different times (0,6,12,24,48 h) and (0,25,50,100 ng / ml) of different concentrations of TRAIL this three cell apoptosis rate The impact (of which 48 hours of flow cytometry 100ng/ml TRAIL role of apoptosis rate). (2) Western blot detection of c-Flip protein expression levels in the three cells. Relatively insensitive (3) cell lines to TRAIL sensitivity and protein expression levels of c-Flip, select cell lines HT29 next experiment, were divided into groups; adding TRAIL (100ng/ml) group; and TRAIL (100ng/ml) group after pretreatment with specific inhibitor ly294002 of,. Were detected by flow cytometry the apoptosis rate changes and changes in c-Flip protein expression detected by Western blot. (4) were stimulated with 100ng/ml TRAIL HT29 cells at different times (0,15,30,60,120 min), Western blot to detect changes in the expression levels of phosphorylated Akt. Results: (1) the SW480 and HCT116 to TRAIL relatively sensitive and showed a dose-dependent, the HT29 relatively insensitive. (2) C-Flip protein expression levels in three cells, and may be related to their different sensitivity to TRAIL. (3) HT29 cells are divided into 3 groups (control group, 100ng/ml the TRAIL, ly294002 100ng/mlTRAIL group) for processing, the results show that c-Flip ly294002 pretreatment group protein expression levels were significantly reduced by flow cytometry The technique detected in ly294002 pretreatment group apoptosis rate increased significantly. (4) HT29 cells basal expression levels of p-Akt 100ng/ml TRAIL HT29 cells 15,30,60,120 min, p-Akt protein expression was significantly reduced, but groups of p-Akt expression was not significant difference. Conclusion: (1) different colon cancer cell lines to TRAIL sensitivity: SW480 and HCT116 relatively sensitive to TRAIL, while the HT29 is relatively insensitive. (2) via the PI3K/Akt pathway can down-regulate the expression of c-Flip, and colon cancer cells HT29 reversed TRAIL sensitivity.
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