Effects of Diyu Huaihua decoction on the proliferation and apoptosis of Human Colorectal Cancer HCT116 Cells by Regulating the p38 MAPK/p53 Pathway
Objective To explore the effects and mechanism of Diyu Huaihua decoction on the proliferation and apoptosis of human colorectal cancer HCT116 cells.Methods HCT116 cells were divided into blank control group(blank serum),positive control group(oxaliplatin-containing serum),Diyu huaihua decoction low-dose,medium-dose and high-dose groups(10%,15%,20%Diyu Huaihua decoction-containing serum).The corresponding drug-containing serum were given for intervention,the cell proliferation activity was detected by MTT.The clone formation ability of each group of cells was detected by the monoclonal formation assay Annexin V-FITC/PI method was used to detecte the level of apoptosis;Hoechst 33258 staining was used to observe cell morphological changes;Western blot was used to detect the protein expression levels of Bax,Bcl-2,cleaved caspase-3,p38MAPK,p-p38MAPK(Thr180/Tyr182),MDM2 and p53 in cells.HCT116 cells were further divided into blank control group(blank serum),Diyu Huaihua decoction group(20%Diyu Huaihua decoction containing serum),p38MAPK inhibitor TAK-715 group(10 mmol·L-1 TAK-715)and combination group(20%Diyu Huaihua decoction containing serum+10 mmol·L-1 TAK-715).The 20%Diyu Huaihua decoction-containing serum and 10 mmol·L-1 TAK-715 were used for intervention.The proliferation activity of cells in each group was detected by MTT;the apoptosis level was detected by Annexin V-FITC/PI method;the protein expression levels of p38MAPK,p-p38MAPK,MDM2 and p53 in the cells were detected by Western blot.Results Compared with the blank control group,the proliferation activity and colony formation ability of HCT116 cells in the Diyuhuaihua decoction medium and high dose groups and the positive control group were significantly decreased(P<0.01).The nucleus showed the changes of shrinking and fragment,the level of apoptosis was significantly increased(P<0.01),and the protein expression levels of Bax/Bcl-2,cleaved caspase-3,p-p38MAPK/p38MAPK and p53 were significantly increased(P<0.05).The protein expression levels of MDM2 were significantly down-regulated(P<0.01).However,combined treatment with TAK-715 could significantly inhibit the growth inhibition and apoptosis induction of HCT116 cells by Diyu Huaihua decoction(P<0.05).Conclusion Diyu Huaihua decoction may inhibit cell proliferation and induce apoptosis in HCT116 cells by regulating the p38 MAPK/p53 pathway.
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