Experimental study in vitro of ziyuglycoside n in inhibition of proliferation,migration,invasion and induction of apoptosis of colon cancer cells
Objective To investigate the effect and its mechanism of ziyuglycoside Ⅱ on prolifer-ation,migration,invasion and apoptosis of colon cancer cells HT-29.Methods The effect of ziyugly-coside n on cell proliferation of colon cancer cells HT-29 was determined by CCK-8 method;the effect of ziyuglycoside Ⅱ on cell migrative capacity of colon cancer cells HT-29 was determined by scratch assay;the effect of ziyuglycoside n on cell invasive capacity of colon cancer cells HT-29 was determined by transwell assay;the effects of ziyuglycoside n on cell apoptosis of colon cancer cells HT-29 was determined by flow cytometry;the effects of ziyuglycoside Ⅱ on mRNA and protein expres-sion of protein kinase B(AKT)/phosphatidylinositol-3-kinase(PI3K)signal pathway were determined by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR)and western-blot,respectively.Results Ziyuglycoside n(0,1,5,10,20,40,60 and 80 μmol/mL)inhibited pro-liferation of colon cancer cells HT-29 in a dose-dependent manner.Ziyuglycoside n(5,10 and 20 μmol/mL)inhibited migration of colon cancer cells HT-29 in a dose-dependent manner.Ziyuglyco-side Ⅱ(5,10 and 20 μmol/mL)inhibited invasion of colon cancer cells HT-29 in a dose-dependent manner.Ziyuglycoside Ⅱ(5,10 and 20 μmol/mL)promoted apoptosis of colon cancer cells HT-29 in a dose-dependent manner.Ziyuglycoside Ⅱ(5,10 and 20 μmol/mL)increased mRNA expression of AKT and PI3K,decreased mRNA expression of Caspase-3 and Caspase-9.Ziyuglycoside Ⅱ(5,10 and 20 μmol/mL)increased protein expression of phosphorylated protein kinase B(p-AKT)and phosphorylphosphatidylinositol-3-kinase(p-PI3K),increased the expression of Cleaved-Caspase-3 and Cleaved-Caspase-9 protein.Conclusion Ziyuglycoside Ⅱ can inhibit proliferation,migration and invasion of colon cancer cells HT-29,and promote the apoptosis of colon cancer cells HT-29.Its mechanism may be related to regulating the signal pathway of AKT/PI3K via promoting phospho-rylation of AKT and PI3K protein,activation of Caspase-3 and Caspase-9 protein.
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