Research on Apoptosis of Human Colon Carcinoma Cell HT-29 Induced by NDGA Combined Celecoxib
|School||Zunyi Medical College,|
|Keywords||nordihyroguaiaretic acid Celecoxib HT29 cells apoptosis Caspase-3|
Objective:We tested the efficacy of the selective cyclooxygenase-2(COX-2)inhibitor Celecoxib,5-lipoxygenase(5-LOX)inhibitor nordihyroguaiaretic acid(NDGA)or their combination on human colon carcinoma cell line HT-29 in vitro, to study the effect of cell proliferation and apoptosis,and the probable mechanism involved.Methods:We used different concentration of Celecoxib and NDGA to dispose cancer cell independently.Used thiazolyl blue tetrazlium bromide(MTT)assay detect cell proliferation.Inverted phase contrast microscope to observe morphologic change of cells.Annexin V/PI fluorescence staining was used to evaluate the apoptotic of HT-29 cells.and reverse transcription polymerase chain reaction(RT-PCR)was used to observe the changes of caspase-3 mRNA expression.The effect of celecoxib and NDGA on the HT-29 of proliferation and related mechanism were studied. Results:The growth of the HT-29 was inhibited by either Celecoxib or NDGA in adose-and time-dependent manner,the effects of combined therapy was better than that of any drug used singly. Under inverted phase contrast microscope, we can observe that cell morphology chaned significantly.and the group of Celecoxib combined NDGA changed most obvious. Apoptosis was observed by laser scanning confocal microscope(LSM)after Celecoxib and NDGA to dispose the HT-29.RT-PCR show that up-regulation of Caspase-3 after drug, and the combination of two drugs increased the most.Conclusion:Either Celecoxib or NDGA can promate the proliferation of colon carcinoma cell line HT-29 while it can inhibit the proliferation and induce apoptosis of HT-29 cells. combined therapy was better than that of any drug used singly.The mechanism may be associated with up-regulation of Caspase-3.