Journal of Jilin University(Medicine Edition) ›› 2022, Vol. 48 ›› Issue (2): 356-363.doi: 10.13481/j.1671-587X.20220212

• Research in basic medicine • Previous Articles     Next Articles

Promotion effect of rutin on apoptosis of human colon cancer SW480 cells and its mechanism

Suxian CHEN1,Zehui GU1,Yangfei MA1,Qi TAN1,Qi LI1,Yadi WANG2()   

  1. 1.Department of Pathology,Third Affiliated Hospital,Jinzhou Medical University,Jinzhou 120001,China
    2.Precision Medicine Center,Third Affiliated Hospital,Jinzhou Medical University,Jinzhou 120001,China
  • Received:2021-06-21 Online:2022-03-28 Published:2022-05-10
  • Contact: Yadi WANG E-mail:15084126408@126.com

Abstract: Objective

To explore the effect of rutin on the colorectal cancer (CRC) SW480 cells, and to clarify the effect of rutin in the treatment of CRC and its possible molecular mechanism.

Methods

The Comparative Toxicogenomics(CTD) and GeneCards databases were used to predict the intersection mRNA related to rutin; Gene Ontdogy(GO) and Kyoto Encyclopedia of Genes and Genones(KEGG) databases were used to predict the biological function and action pathways of the intersection mRNA; String database was used to find the Notch-1 related proteins. The human CRC SW480 cells were used as the subjects, 0.1, 1.0, and 10.0 μmol·L-1 DAPT were given for preliminary screening, then 1, 2, 4, 8 and 10 μmol·L-1 DAPT were given to intervene, and the optimal concentrations of rutin and DAPT were screened out. The human CRC SW480 cells were divided into blank control group, rutin group (22 μmol·L-1), DAPT group (1 μmol·L-1) and rutin+DAPT group(22 μmol·L-1 rutin+1 μmol·L-1 DAPT).MTT method was used to detect the cell proliferation activities; Hoechst33258 nuclear staining was used to observe the apopotic morphology of cells,and flow cytometry was used to detect the apoptotic rates;real-time fluorescence quantitative PCR(RT-qPCR) method was performed to detect the expression levels of Notch-1 mRNA in the SW480 cells in various groups; Western blotting method was used to determine the protein expression levels of Notch-1,caspase-3,caspase-9,B-cell lymphoma 2(Bcl-2),and Bcl-2 associated X protein in the SW480 cells in various groups.

Results

In CTD and GeneCards databases, there were a total of 34 intersection mRNA;the GO and KEGG databases predicted that the intersection mRNA was related to the process of colon cancer apoptosis; String database verified 16 Notch-1 related proteins (score>0.42); the optimal administration concentrations of rutin and DAPT were 22 μmol·L-1 and 1 μmol·L-1; compared with blank control group, the apoptotic rates in rutin group, DAPT group and rutin+DAPT group were increased significantly (P<0.05), the expression levels of Notch1 mRNA and protein were significantly reduced (P<0.05), the expression levels of caspase-3, caspase-9, and Bax proteins in the cells were increased significantly(P<0.05), while the expression levels of Bcl-2 protein were decreased significantly (P<0.05). There was no significant difference in the apoptotic rate between rutin group and DAPT group (P>0.05). Compared with rutin group and DAPT group, the apoptotic rate in rutin+DAPT group was significantly increased(P<0.05), the expression levels of caspase-3, caspase-9, and Bax proteins were increased significantly(P<0.05),while the expression level of Bcl-2 protein was decreased significantly(P<0.05).

Conclusion

The results of bioinformatics analysis and experiment show that rutin can promote the apoptosis of CRC cells, which may be achieved by targeting Notch-1 and regulating MAPK signal pathway.

Key words: Rutin, DAPT, Colon neoplasms, Apoptosis, Mitogen-activated protein kinase, Notch-1

CLC Number: 

  • R735.35