Apoptotic Mechanism in Human HT-29 Colon Cancer Cells Induced by Total Flavones from Trollius chinensis Bunge
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Abstract:
The mechanism of the ethanol extract of the total flavones from Trollius chinensis Bunge (TFETC) inducing apoptosis in HT-29 human colon cancer cells was investigated. MTT assay, which is used to determine the anti-proliferation effect to cancer cells, showed that TFETC inhibited the proliferation of HT-29 cells in a dose-dependent manner, and the inhibitory rate was 81% when treated with 200 μg/mL TFETC. 4,6-diamidino-2-phenylindole (DAPI) staining. Flow cytometry analysis showed that 200 μg/mL TFETC induced apoptosis of HT-29 cells and the sub-G1 DNA content was 28.9%. Reverse transcription polymerase chain reaction (RT-PCR) was applied to evaluate the impact of TFETC on mRNA expression levels of Bcl-2, Bcl-xL, Bax, caspase-9, caspase-3 and COX-2 in HT-29 cells. The results showed that TFETC reduced the mRNA expression levels of anti-apoptosis genes Bcl-2 and Bcl-xL, and up-regulated the mRNA expression of pro-apoptosis genes Bax, caspase-9, and caspase-3 in HT-29 cells. Moreover, TFETC down-regulated the mRNA expression of COX-2 in HT-29 cells in a dose-dependent manner. These results indicated that TFETC could induce the apoptosis through an intrinsic mitochondria pathway in HT-29 cells. Meanwhile, TFETC could reduce cell proliferation through down-regulating the gene expression of COX-2.
