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Gynura procumbens modulates the microtubules integrity and enhances distinct mechanism on doxorubicin and 5-flurouracil-induced breast cancer cell death

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Abstract

Recent studies both in vitro and in vivo of G. procumbens exhibits chemopreventive properties for tumor inhibition on several types of cancer. Our study was carried out to observe the anticancer property of ethyl acetate fraction of G. procumbens leaves (FEG) on breast cancer cells as well as the co-chemotherapeutic potential, and to investigate its molecular mechanisms. 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to measure the growth inhibitory effect of FEG, doxorubicin (DOX), and 5-fluorouracil (5-FU) and their combination. Flowcytometry, 4′,6-diamidino-2-phenylindole (DAPI) staining, and immunobloting were used to explore the mechanism of cell cycle arrest and apoptosis. FEG inhibited cell proliferation, induced G1 phase arrest and apoptosis. The inhibitory effect of FEG was enhanced when combined with Dox and 5-FU. The apoptosis induction was related to the increase of c-PARP expression after combination treatment of FEG and Dox or 5-FU on MCF-7 cells. However, treatment of DOX, 5-FU, and FEG on T47D cells, resulting no significance DNA fragmentation and nuclei condensation evidance. Only combination treatment of 5-FU + FEG showed c-PARP expression in T47D cells. In T47D cells, The FEG treatment also caused the decrease of microtubule expression as shown by Western blotting assay. The decreasing level of microtubul expression might be caused by protein aggregation, as shown by immunostaning using α-tubulin antibody. All these results suggest that FEG potentiates the DOX and 5-FU efficacy on MCF-7 and T47D cells. FEG induces T47D cell death through different mechanism than MCF-7 that proposed to be mitotic catastrophe. The FEG may have specific targeted on microtubule integrity modulation leading to the cell cycle arrest and proliferation inhibition. Further FEG could be developed as a co-chemotherapeutic agent for reducing side effect and have specific molecular target for breast cancer.

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Acknowledgement

This work was supported by Program Hibah Bersaing 2010 and Sandwich-like program 2010/2011, managing by Directorate General of Higher Education Republic of Indonesia, 2010

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Authors and Affiliations

  1. Graduate Program of Pharmaceutical Science, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia

    Nunuk Aries Nurulita

  2. Faculty of Pharmacy, Universitas Muhammadiyah Purwokerto, Purwokerto, Central Java, Indonesia

    Nunuk Aries Nurulita

  3. Cancer Chemoprevention Research Center, Faculty of Pharmacy, Universitas Gadjah Mada, Sekip Utara, Yogyakarta, 55281, Indonesia

    Edy Meiyanto &  Sugiyanto

  4. Departement of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia

    Sugiyanto

  5. Laboratorium of Gene Function of Animal, Nara Institute of Science and Technology, Takayama, Ikoma, Japan

    Eishou Matsuda & Masashi Kawaichi

Authors
  1. Nunuk Aries Nurulita
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  2. Edy Meiyanto
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  3. Sugiyanto
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  4. Eishou Matsuda
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  5. Masashi Kawaichi
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Corresponding author

Correspondence to Edy Meiyanto.

Additional information

Nunuk Aries Nurulita has made conception and design of this study, acquisition of data, data collection, analysis and interpretation and statistical data, and drafted the manuscript. Edy Meiyanto and Sugiyanto have made conception and design of this study, analysis and interpret data, and drafted the manuscript. Eishou Matsuda and Masashi Kawaichi have interpreted data and reviewed the manuscript. All author have already read and approved the final revision of this manuscript.

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Nurulita, N.A., Meiyanto, E., Sugiyanto et al. Gynura procumbens modulates the microtubules integrity and enhances distinct mechanism on doxorubicin and 5-flurouracil-induced breast cancer cell death. Orient Pharm Exp Med 12, 205–218 (2012). https://doi.org/10.1007/s13596-012-0063-5

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