Effect of celastrus orbiculatus extract on cell biological behaviors in colon cancer by regulating microRNA-378
Objective To explore the effect of celastrus orbiculatus extract on the biological behaviors of colon cancer cells Caco-2 and its possible mechanism.Methods Human colon cancer cells Caco-2 were induced with low-dose(20 pg/L),medium-dose(60 μg/L),and high-dose celastrus orbiculatus extract(180 μg/L),30 μmol/L cyclophosphamide(positive control)and blank control.In addition,Caco-2 cells either treated with blank control or high-dose celastrus orbiculatus extract were transfected with miR-NC,miR-378 mimics,anti-miR-NC and anti-miR-378.Nine replicates were set per group.Cell proliferation and apoptosis were examined by MTT assay and flow cytometry,respectively.Cell migration and invasion were detected by Transwell assay.were used to treat.Protein expressions of Ki67,matrix metalloproteinase-2(MMP-2),Cleaved-caspase-3,and MMP-9 were determined by Western blot.The mRNA level of miR-378 was detected by reverse transcription quantitative real-time polymerase chain reaction(qRT-PCR).Results Low-dose,medium-dose and high-dose Celastrus orbiculatus extract significantly decreased cell proliferation rate,protein levels of Ki67,MMP-2,and MMP-9,and number of migrating and invading cells(P＜0.05),but significantly increased the expression of miR-378,apoptosis rate and protein level of Cleaved-caspase-3 in a dose-dependent manner(P＜0.05).After transfection of miR-378 mimics,the cell proliferation rate,protein levels of Ki67,MMP-2 and MMP-9,and number of migrating and invading cells significantly decreased(P＜0.05),and the apoptosis rate,the expression of Cleaved-caspase-3 and miR-378 significantly increased than those transfected with miR-NC(P＜0.05).In comparison to those transfected with anti-miR-NC and induced with high-dose Celastrus orbiculatus extract,transfection of anti-miR-378 in cells induced with high-dose Celastrus orbiculatus extract significantly increased cell proliferation rate,protein levels of Ki67,MMP-2,and MMP-9,and number of migrating and invading cells(P＜0.05),but significantly decreased the expression of miR-378,apoptosis rate and protein level of Cleaved-caspase-3(P＜0.05).Conclusion Celastrus orbiculatus extract could cell biological behaviors in colon cancer by upregulating miR-378.
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