XL413 inhibits colon cancer cell proliferation by inhibiting DNA replication and inducing cell apoptosis

Abstract:

Objective To investigate the effect of XL413，a drug inhibiting cell division cycle 7（CDC7），on colorectal cancer cells and its mechanism of action.Methods Multiple colorectal cancer cell lines were treated with normal saline and XL413 at different concentrations for 72 hours. CCK-8 assay was used to measure the absorbance of cells at 450 nm and calculate cell viability and the half-maximal inhibitory concentration（IC₅₀） of XL413；Annexin V-FITC/PI double staining was used to measure cell apoptosis；colony formation assay was used to observe the regenerative ability of cells；RNA-seq was used to observe the effect of XL413 on cell transcriptome；qRT-PCR was used to measure the relative mRNA expression levels of apoptosis-related genes；Western blot was used to measure the phosphorylation level of minichromosome maintenance protein 2（MCM2），a member of the DNA unwinding minichromosome maintenance protein complex family，and the expression levels of apoptosis-related proteins.Results Compared with the control group，the XL413 treatment group had significant reductions in the abilities of colorectal cancer cell proliferation，migration，and colony formation（P=0.001 5） and a significant increase in the proportion of cells in the early and late stages of apoptosis（P=0.000 6）. As for the transcriptional expression level of genes，the XL413 treatment group had a significant reduction in the ratio of the cell apoptosis markers B-cell lymphoma-2（Bcl-2） to Bcl-2-associated X（P=0.008 7），and at the protein level，XL413 inhibited the phosphorylation of the DNA unwinding-related protein MCM2 and thus DNA replication and promoted the increase in the expression of apoptosis-related proteins.Conclusion XL413 can inhibit DNA replication by reducing the phosphorylation of the DNA unwinding-related protein MCM2 and induce the apoptosis of colorectal cancer cells. The study provides a theoretical basis for the application of XL413 in the treatment of colorectal cancer in the future.