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SCIENCE CHINA Life Sciences, Volume 60, Issue 4: 417-428(2017) https://doi.org/10.1007/s11427-016-0368-y

Lycorine inhibits breast cancer growth and metastasis via inducing apoptosis and blocking Src/FAK-involved pathway

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  • ReceivedNov 29, 2016
  • AcceptedDec 27, 2016
  • PublishedFeb 27, 2017

Abstract

Breast cancer is the most commonly diagnosed cancer type worldwide among women and more than 90% of patients die from tumor metastasis. Lycorine, a natural alkaloid, has been widely reported possessing potential efficacy against cancer proliferation and metastasis. In our study, the anti-tumor potency on breast cancer was evaluated in vitro and in vivo for the first time. Our results indicated that lycorine inhibited breast cancer cells growth, migration and invasion as well as induced their apoptosis. In in vivo study, lycorine not only suppressed breast tumor growth in xenograft models and inhibited breast tumor metastasis in MDA-MB-231 tail vein model. More importantly, we found lycorine had less toxicity than first-line chemotherapy drug paclitaxel at the same effective dose in vivo. Furthermore, on mechanism, lycorine inhibited tumor cell migration and invasion via blocking the Src/FAK (focal adhesion kinase)-involved pathway. In conclusion, our study implied lycorine was a potential candidate for the treatment of breast cancer by inhibition of tumor growth and metastasis.


Funded by

National Natural Science Foundation of China(81272463,81472788)

Major State Basic Research Development Program of China(2015CB910400)


Acknowledgment

This work was supported by National Natural Science Foundation of China (81272463, 81472788), and Major State Basic Research Development Program of China (2015CB910400).


Interest statement

The author(s) declare that they have no conflict of interest. All procedures performed in studies involving animals were in accordance with the ethical standards of the animal investigation committee of the Institute of Biomedical Sciences, East China Normal University.


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  • Figure 1

    Lycorine inhibits the proliferation of breast cancer cells. A, Chemical structure and molecular weight of lycorine. B, Lycorine inhibited the growth of breast cancer cells and less cytotoxicity was observed on MCF-10A, L02 and HAF cell lines. C, Lycorine inhibited the clone formation of breast cancer cells. Corresponding statistical results which were represented the average of three replications were shown. (*, P<0.05; **, P<0.01; ***, P<0.001 vs. control).

  • Figure 2

    (Color online) Lycorine inhibits the migration and invasion of breast cancer cells. A, Lycorine inhibited cell migration of 4T1 and MDA-MB 231 in wound-healing assay. Images of migrated cells were captured and dotted lines indicated the field of initial scraping. Lycorine inhibited 4T1 and MDA-MB-231 migration (B) and invasion (C) in transwell chamber assay. Three independent experiments were performed (*, P<0.05; **, P<0.01; ***, P<0.001 vs. control).

  • Figure 3

    Lycorine prevents breast tumor growth in MDA-MB-231 subcutaneous xenograft mouse model. A, MDA-MB-231 cells were injected subcutaneously into female nude mice. After 30 days, tumors were removed and photographed. B, The weight of removed tumors. C, Chang of tumor volume with the treatments of different agents. D, Chang of mice body weight with the treatments of different agents. E, Haematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) analysis of tumors. Arrows showed PCNA in the tumor, with the statistical result on the right (*, P<0.05; **, P<0.01; ***, P<0.001 vs. control).

  • Figure 4

    Lycorine prevents breast tumor growth in 4T1 orthotopic xenograft mouse model. A, Female nude mice received orthotropic injection in the No.4 fourth mammary fat pad with 4T1 cells. After 21 days, primary tumors were removed and photographed. B, The weight of removed tumors. H&E (C) staining and IHC (D) analysis of tumors and organs. Arrows showed PCNA in the tumor, with the statistical result on the right. (*, P<0.05; **, P<0.01; ***, P<0.001 vs. control).

  • Figure 5

    Lycorine suppressed breast tumor pulmonary metastasis in mice. A, MDA-MB-231-Luc cells were injected into lateral tail vein. Bioluminescence images were taken on the 21st day before mice scarified. Representative images and statistical result were shown. B, Lungs were removed and tumor nodules on surface were counted and photographed. Statistical result were shown on the right. C, H&E staining analysis of lungs and the statistical result was shown on the right. (*, P<0.05; **, P<0.01; ***, P<0.001 vs. control).

  • Figure 6

    (Color online) Lycorine induced apoptosis and blocked Src/FAK/MMP signaling pathway. A, The effect on apoptosis of breast cancer cells which were treated with lycorine at the indicated concentration for 48 h. B, Statistical result of cell apoptosis induced by lycorine. C, The expression of PARP, CL. PARP, Caspase3, CL. Caspase3, Bax and Bcl-2 were detected by western blot analysis in 4T1 and MDA-MB-231 after incubated with indicated concentration of lycorine for 48 h. D, The expression of p-FAK, p-Src, Src, p-JNK, JNK, p-c-Jun, c-Jun and MMP2 were determined by western blot analysis in 4T1 and MDA-MB-231 after incubated with indicated concentration of lycorine for 24 h.

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