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Effects and Mechanism of Cantharidin on Apoptosis of Breast Cancer Cell Line MCF-7

Author: LiXiuZuo
Tutor: GaoYaJie
School: Dalian Medical University
Course: Oncology
Keywords: Cantharidin Breast neoplasms Apoptosis
CLC: R737.9
Type: Master's thesis
Year: 2011
Downloads: 15
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Abstract


Objective: The incidence rate of breast cancer in our country increased year by year, which became the most common malignant tumor in women and has leapt to the top of women cancer. Mylabris has been used as a traditional Chinese medicine. The active constituent of mylabris is cantharidin.In recent studies, cantharidin have shown strong in vitro antitumor activity against the cells of various cancers such as colorectal , hepatoma, bladder carcinoma cell.The aim of the research is to observe the effects of cantharidin on apoptosis of breast cancer cell line MCF-7 in vitro and its mechanism of action discussed .Methods: To deal the logarithmic phase of growth human breast cancer cell line MCF-7 treated with different concentration of cantharidin for 48 hours , MTT assay was used to detect OD value and calculate inhibition ratio. The apoptotic ratio and cell cycle was analyzed by flow cytometry. The expressiona of Bcl-2 and Bax were determined by immunofluorescence method.Results:1.MTTThe inhibition ratio of human breast cancer cell line MCF-7 increased, when dentisy of cantharidin increased. The inhibition ratio of the maximum is 68%, when the dentisy was 100μmol/L, IC50 is 30.5μmol/L.2. Flow cytometry resultsThe cell apoptotic ratio of 12.5μmol/L、25μmol/L、50μmol/L density was 27.91%、36.12%、63.53% by quantitative analysis of marked AnnexinⅤ-FITC and flow cytometry. Compared with control group(0.87%), the cell apoptotic ratio was significance the differences from the respective control groups(P﹤0.05). Apoptotic peak appeared in DNA bargraph of flow cytometry. With the concentration of cantharidin increased, the number of the cells in G1 phase descended gradually, on the contrary , S phase increased gradually.3. Immunofluorescence results With the concentration of cantharidin increased, immunofluorescence intensity of Bcl-2 become weaker gradually, while immunofluorescence intensity of Bax increased gradually.Conclusions: Cantharidin has significant growth inhibiting effect and inducing apoptosis on human breast cancer cell line MCF-7. The apoptosis mechanisms probably was concerned with down-regulation of Bcl-2 and up-regulation of Bax in cell lines at protein level.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Breast tumor
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