Dissertation > Medicine, health > Oncology > Gastrointestinal Cancer > Intestinal neoplasms > Colon tumor

Study of Berberine’s Effect on the Apoptosis and Migration of Colon Cancer Cells

Author ChenBinBin
Tutor HuTianHui
School Xiamen University
Course Microbiology
Keywords colon cancer berberine apoptosis and migration
CLC R735.35
Type Master's thesis
Year 2014
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Colon cancer is one of common malignant digestive tumors from human, and it is a serious threat to human health. Colon cancer can be treated by surgical resection, however, more than half will appear the recurrence and metastasis after treatment. Thus, to seek an effective anticancer drugs was one of the key issues in colon cancer therapy. Berberine, a kind of alkaloid, is extracted from rhizoma coptidis, cortex phellodendri and other traditional Chinese medicines, and clinically for the treatment of gastrointestinal digestive tract disease for a long time. Studies show that berberine has extensive pharmacological action, and it gradually gets attention and recognition by people in antitumor pharmacological value in recent years. What’s more, berberine is widely appreciated because of many advantages such as low price, little toxic or side effects and rich medicine source. Studies prove that berberine has inhibitory effect on many kinds of tumor cells, but the mechanism has not been thoroughly studied.In this paper, the human colon cancer cell lines KM12C, KM12SM and KM12L4A are regarded as the research object, are treated with berberine, to investigate the effect of berberine on apoptosis and migration and its molecular mechanism in human colon cancer cells. In the aspect of apoptosis, firstly, we determined the viabilities of the three human colon cancer cell lines treated by different concentration of berberine. The results show that berberine can significantly inhibits the viabilities of human colon cancer cell lines KM12C, KM12SM and KM12L4A in a dose-dependent manner. Next, we use Hoechst33342staining to observe the morphological changes of the three kinds of human colon cancer cell lines, and the apoptosis rate was detected with the Annexin-V/PI staining assay. In addition, the mitochondrial membrane potential changes are detected by flow cytometry instrument to investigate whether berberine can induce human colon cancer cells apoptosis. We found that berberine can effectively induce human colon cancer cell lines KM12C, KM12SM and KM12L4A apoptosis in a dose-dependent manner as well. In the meantime, we analyzed the proteins of KM12C cells induced by berberine treatment with Western Blotting assay to further clarify the apoptotic molecular mechanisms of human colon cancer cells. The results show that berberine can decrease the expression of Bcl-2, Caspase-3,-9, PARP, and make PARP activated in shear.In addition, the expression of Bax and AIF are increased by berberine treatment, and AIF, Cyt c was released from mitochondria. Moreover, we find that berberine can up-regulate the expression of Fas and p53, or down-regulate p65, Ras, p38, Akt and β-catenin. Finally, we establish a nude mice model of human colon cancer cells to verify berberine can inhibit the growth and proliferation of KM12C cells in vivo. In another aspect of migration, We use wound healing assay, Immunofluorescence and western blot to prove that berberine can effectively suppress the effect of migration in human colon cancer cells and effect on the localization of Exo70. And the inhibitory action may be connected with the down-regulation of MMP-2,-9and Exo70which involved in migration.

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