Apigenin up-regulates expression of miR-34a-5p and enhances sensitivity to fluorouracil in MHCC97H-derived spheres
Objective To determine the effect of apigenin(API)on the sensitivity of human hepatocellular carcinoma MHCC97H-derived spheres(MH-SFC)to 5-fluorouracil(5-FU),and related mechanism.Methods MH-SFC were obtained from MHCC97H cell line by sphere formation assay with serum-free stem cell medium in ultra low attachment plates.The IC50 of 5-FU or API on the cell viability in MH-SFC and MHCC97H cells was assessed by cell counting kit-8(CCK-8)assay.MiR-34a-5p expression level was measured by qRT-PCR.After pre-treatment with API(10.0 and 40.0 μmol·L-1)or co-treatment with API(40.0 μmol·L-1)and miR-34a-3p inhibitor(Anti-34a),IC50 of 5-FU was determined in MH-SFC.The expressions of FoxM1 and c-Myc protein were analyzed by Western blot.Results IC50 of 5-FU was elevated in MH-SFC compared with MHCC97H cells.API preferentially inhibited the cell viability of MH-SFC.IC50 of 5-FU was reduced in MH-SFC by pre-treatment of API(10.0 and 40.0 μmol·L-1).Compared with the MHCC97H cells,MH-SFC had lower expression of miR-34a-5p as well as API up-regulated the expression of miR-34a-5p.Anti-34a rescued the effect of API on the sensitivity to 5-FU and expression of miR-34a-5p in MH-SFC.In addition,API(10.0 and 40.0 μmol·L-1)down-regulated the expressions of FoxM1 and c-Myc protein in MH-SFC.Anti-34a decreased the down-regulation of expressions of FoxM1 and c-Myc protein in MH-SFC by API.Conclusion API can enhance the sensitivity of MH-SFC to 5-FU,the mechanism may be associated with up-regulating miR-34a-5p expression,and subsequently interrupting FoxM1/c-Myc signaling pathway.
NETL
NSTL
万方数据
