Mechanism of polysaccharide of Farfarae Flos in inhibiting proliferation,migration,and invasion of cervical cancer cells by regulating miR-4282 expression
Objective To explore the mechanism of polysaccharide of Farfarae Flos in inhibiting the proliferation,migration,and invasion of cervical cancer cells by regulating miR-4282 expression.Methods Human cervical cancer cell lines HeLa and C-33A were cultured in vitro and treated with 10,20,40,80,and 120 mg·L-1 of polysaccharide of Farfarae Flos.CCK-8 assay was used to determine the survival rate of each group of cells and screen the optimal concentration of polysaccharide of Farfarae Flos.HeLa and C-33A cells were randomly grouped into control group,fluorouracil(5-FU)group,polysaccharide of Farfarae Flos group,miR-4282 mimics group,negative control group,and polysaccharide of Farfarae Flos+miR-4282 inhibitor group.The miR-4282 expression was examined by RT-PCR;the cell proliferation and apoptosis were deterrmined by CCK-8 assay,EDU assay,and flow cytometry;the cell migration and invasion were checked by scratch and transwell assays;immunoblotting was used to examine the expression of cell proliferation related proteins(Cyclin D1),apoptosis related proteins(Bax,Bcl-2),epithelial mesenchymal transition-related proteins[Vimentin,Matrix Metallo-proteinases 2(MMP2),E-cadherin].Results Compared with the control group,the cell survival rate,EDU positivity rate,migration rate,migration number,invasion number,the Cyclin D1 protein expression as well as protein expressions of Bcl-2,Vimentin,and MMP-2 in 5-FU group,polysaccharide of Farfarae Flos group,and miR-4282 mimics group decreased(P＜0.05),while the miR-4282 expression,apoptosis rate,and protein expressions of Bax and E-cadherin increased(P＜0.05).There was no significant change in various indicators of cells in the negative control group(P＞0.05).Compared with the polysaccharide of Farfarae Flos group,the cell survival rate,EDU positivity rate,migration rate,migration number,invasion number,the Cyclin D1 protein expression as well as protein expressions of Bcl-2,Vimentin,and MMP-2 in the polysaccharide of Farfarae Flos+miR-4282 inhibitor group were higher(P＜0.05),while the miR-4282 expression,apoptosis rate,and protein expressions of Bax and E-cadherin were lower(P＜0.05).Conclusion The polysaccharide of Farfarae Flos can inhibit the proliferation,migration,invasion,and apoptosis of cervical cancer cells by up-regulating the miR-4282 expression,ultimately playing a significant role in cancer suppression.
polysaccharide of Farfarae FlosmiR-4282 expressioncervical cancerproliferationmigrationinvasion
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