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Portulaca Oleracea Extract Can Inhibit Nodule Formation of Colon Cancer Stem Cells by Regulating Gene Expression of Notch Signal Transduction Pathwa

Posted on:2017-07-04Degree:Ph.DType:Dissertation
University:Michigan Technological UniversityCandidate:Chen, LiFull Text:PDF
GTID:1474390017463746Subject:Biochemistry
Abstract/Summary:
Purpose: The main aim of this study was to investigate the effects of Portulaca oleracea extract on tumor formation in colon cancer stem cells and chemotherapy sensitivity. In addition, this study analyzed the genetic changes within the Notch signal transduction pathway associated with the effects of the extracts.;Methods: Serum-free cultures of colon cancer cells (HT-29) and HT-29 stem cells were treated with the chemotherapeutic drug 5-FU to assess sensitivity. Injections of the stem cells were also given to BALB/c mice to confirm tumor growth and characteristics. In addition, the effect of different concentrations of P. oleracea extract was tested on the growth of HT-29 and HT-29 stem cells, which was determined by the MTT method. The effects of P. oleracea extract on the expression of beta-catenin, Notch1, and Notch2 in HT-29 cells were detected by RT-PCR and Western blotting.;Results: The tumor volume of HT29 cells was two times larger than that of HT29 stem cells. Treatment with P. oleracea extracts inhibited the proliferation of both HT-29 and HT-29 stem cells in doses from 0.07 - 2.25 mug/mL. Apoptosis of HT-29 and HT-29 stem cells, as assessed by flow cytometry, was enhanced by the addition of P. oleracea extract. Finally, treatment with P. oleracea extracts significantly down-regulated the expression of the Notch1 and beta-catenin genes in both cell types.;Conclusion: This study shows that P. oleracea extract inhibits the growth of colon cancer stem cells in a dose-dependent manner. Furthermore, it inhibits the expression of the Notch1 and beta-catenin genes. Taken together, this suggests that it may elicit its effects through regulatory and target genes that mediate the Notch signal transduction pathway.
Keywords/Search Tags:Oleracea extract, Notch signal transduction, Stem cells, Effects, Expression
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