Effect of Leonurine on proliferation,apoptosis,and chemotherapy resistance of colon cancer cells by regu-lating the Hippo-YAP signaling pathway
Objective To investigate the effects of Leonurine(Leo)on the proliferation,apoptosis,and chemotherapy resistance of colon cancer cells by regulating the Hippo-Yes associated protein(YAP)signaling path-way.Methods Human colon cancer LOVO cells were treated with Leo at concentrations of 0,1.875,3.75,7.5,15,30,and 60 μg/mL for 48 hours,respectively,and suitable Leo concentrations were selected for the experiment.LOVO cells were separated into blank group,Leo low-dose group,Leo medium-dose group,Leo high-dose group,Leo high-dose+pcDNA group,and Leo high-dose+pcDNA-YAP group,CCK-8 method and 5-ethynyl-2'-deoxyuridine(EdU)staining were applied to detect cell proliferation;flow cytometry was applied to detect cell apoptosis;qRT-PCR was applied to detect the mRNA expression of proliferating cell nuclear antigen(PCNA),Bc12 associated X protein(Bax),multidrug resistance associated protein 1(MRP1),and P-glycoprotein(PGP)in LOVO cells;Western blot was applied to detect the expression of YAP,connective tissue growth factor(CTGF),and cysteine rich protein 61(CYR61)in LOVO cells.LOVO/pentafluorouracil(5-FU)cells were studied and treated according to the above groups.They were divided into A-blank group,A-Leo low-dose group,A-Leo medium dose group,A-Leo high-dose group,A-Leo high-dose+pcDNA group,A-Leo high-dose+pcDNA-YAP group.CCK-8 was applied to de-tect the drug resistance of LOVO/5-FU cells.Results Compared with the blank group,the A450 value of LOVO cells(0.86±0.08,0.75±0.06,0.43±0.04 vs 0.97±0.08),EdU positive cell rate(50.96％±2.17％,43.37％±2.05％,28.84％±1.19％vs 57.75％±2.36％),the expression of PCNA,MRP1,PGP mRNA,and the expression of YAP(2.05±0.17,1.78±0.14,1.23±0.11 vs 2.36±0.19),CTGF,CYR61 proteins in the Leo low dose group,Leo medium dose group,and Leo high dose group decreased,while the apoptosis rate(15.54％±0.65％,23.38％±1.01％,36.69％±1.78％vs 8.34％±0.31％)and the expression of Bax mRNA increased,in a dose-dependent manner(P＜0.05);compared with the Leo high-dose group and Leo high-dose+pcDNA group,the A450 value of LOVO cells(0.77±0.05 vs 0.43±0.04,0.45±0.04),EdU positive cell rate(46.69％±1.95％vs 28.84％±1.19％,27.67％±1.25％),the expression of PCNA,MRP1,PGP mRNA,and the expression of YAP,CTGF,CYR61 proteins in the Leo high-dose+pcDNA-YAP group increased,while the apoptosis rate and the expression of Bax mRNA de-creased(P＜0.05);compared with the A-blank group,the LOVO cell viability of the A-Leo low-dose group,A-Leo medium dose group,and A-Leo high-dose group decreased,in a dose-dependent manner(P＜0.05);compared with the A-Leo high-dose group and the A-Leo high-dose+pcDNA group,the LOVO cell viability of the A-Leo high-dose+pcDNA-YAP group increased(P＜0.05).Conclusion Leo may inhibit proliferation and chemotherapy resistance of colon cancer cells and promote cell apoptosis by inhibiting the Hippo-YAP signaling pathway.
LeonurineColon cancerHippo-Yes associated protein signaling pathwayProlifera-tionApoptosisChemotherapy resistance
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维普
