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Effects of 5-Aza-CdR on the proliferation of human breast cancer cell line MCF-7 and on the expression of Apaf-1 gene

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Summary

Hypermethylation in the promoter region of tumor suppressor genes is a common mechanism of gene silencing, which tends to occur in cancer. The effects of 5-Aza-2′-deoxycytidine (5-Aza-CdR), a specific DNA methyltransferase inhibitor, on the cell proliferation of human breast cancer cell line MCF-7 and on the expression of Apaf-1 gene were investigated. Human MCF-7 cells were incubated with increasing concentrations of 5-Aza-CdR for 12 to 120 h. The growth inhibition rates of MCF-7 cells were detected by MTT assay. Changes of cell cycle distribution and apoptotic rates of MCF-7 cells were determined by flow cytometry. The expressions of DNA methyltransferase 3b mRNA and Apaf-1 mRNA were measured by reverse transcription polymerase chain reaction (RT-PCR). Meanwhile, the expression of Apaf-1 protein was detected by Western blotting. The results showed that 5-Aza-CdR significantly inhibited the growth of MCF-7 cells and the growth inhibition rate of MCF-7 cells was significantly enhanced with the concentration of 5-Aza-CdR and the action time. Flow cytometry indicated that 5-Aza-CdR could significantly induce G1/S cell cycle arrest and increase the apoptosis rate of MCF-7 cells. The mRNA and protein expressions of Apaf-1 were up-regulated in MCF-7 cells treated with 5-Aza-CdR, which was accompanied by down-regulation of DNA methyltransferase 3b mRNA. It is concluded that 5-Aza-CdR might retard the growth of tumor cells and promote the apoptosis of MCF-7 breast cancer cells by inhibiting the expression of DNA methyltransferase 3b and re-activating the Apaf-1 gene expression.

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Authors and Affiliations

  1. Cancer Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Huihua Xiong  (熊慧华), Hong Qiu  (邱 红), Liang Zhuang  (庄 亮), Hua Xiong  (熊 华) & Yuan Chen  (陈 元)

  2. Cancer Center, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China

    Rui Jiang  (姜 蕊)

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  1. Huihua Xiong  (熊慧华)
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  2. Hong Qiu  (邱 红)
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  3. Liang Zhuang  (庄 亮)
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  4. Hua Xiong  (熊 华)
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  5. Rui Jiang  (姜 蕊)
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  6. Yuan Chen  (陈 元)
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Xiong, H., Qiu, H., Zhuang, L. et al. Effects of 5-Aza-CdR on the proliferation of human breast cancer cell line MCF-7 and on the expression of Apaf-1 gene. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 29, 498–502 (2009). https://doi.org/10.1007/s11596-009-0421-9

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  • DOI: https://doi.org/10.1007/s11596-009-0421-9

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