Induction Effect and Underlying Mechanism of Scutellarin on Apoptosis of Human Colorectal Cancer SW480 Cells and AOM/DSS Mouse
Abstract Background: This study was aimed to investigate the effect of Scutellarin on apoptosis of human colorectal cancer SW480 cells and azoxymethane (AOM)/dextran sulfate sodium (DSS) mouse, and clarify its mechanism. Methods: 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT) and cell migration assay were performed to detect the viability and proliferation of SW480 cells that treated with different concentrations of Scutellarin. Hoechst33342 Staining to determine apoptosis, detecting Caspase-3 and Caspase-9 activities after administration of different concentrations of Scutellarin. AOM/DSS mouse were administrated with Scutellarin; immunohistochemistry and western blot was employed to detect the effect on the expression of BCL2-Associated X (Bax) and B-cell lymphoma-2 (Bcl-2) proteins, and the message Ribonucleic Acid (mRNA) levels of Bax, Bcl-2, Caspase-3and Caspase-9 were assessed with quantitative reverse transcription polymerase chain reaction (qRT-PCR). The data was analyzed by one-way analysis of variance (ANOVA) with SPSS19.0 software, expressed as mean ± standard deviation (x ± s). Results: Scutellarin could inhibit the proliferation of SW480 cells after treatment with different concentrations of Scutellarin, while the chromatin condensation and nucleus showed more intense blue fluorescence. Moreover, Scutellarin could significantly increase the activities of Caspase-3 and Caspase-9 as well as different concentrations of Scutellarin could significantly down-regulate the expression of apoptosis related protein Bcl-2 and up-regulate the expression of Bax protein. When compared with the solvent control group, the relative expression levels of the related gene Bcl-2 were down-regulated by different concentrations of Scutellarin, while the Bax, Caspase-3 and Caspase-9 genes were up-regulated. Conclusion: Scutellarin can inhibit the in vitro activity of colon cancer SW480 cells, promote Bax expression, inhibit Bcl-2 expression, and up-regulate the activities of apoptotic enzymes Caspase-3 and Caspase-9 to induce apoptosis in vivo and in vitro, suggesting that it has a certain therapeutic effect on colon cancer.