Science China Life Sciences

, Volume 61, Issue 5, pp 550–558 | Cite as

Regulatory effects of antitumor agent matrine on FOXO and PI3K-AKT pathway in castration-resistant prostate cancer cells

  • Qi Li
  • Hai Huang
  • Zheng He
  • Yi Sun
  • Yufeng Tang
  • Xiaohong Shang
  • Chengbin WangEmail author
Research Paper


We previously demonstrated that matrine could inhibit the proliferating, migrating, as well as invading processes of both PC-3 and DU145 cells. However, the underlying molecular mechanisms have not yet been clearly defined. In this study, using various techniques such as high throughput sequencing technology, bioinformatics, quantitative real-time PCR, and immunoblot analysis, we aimed to understand whether matrine serves as a novel regulator of FOXO and PI3K-AKT signaling pathway. DU145 and PC-3 cell lines were cultured for 24 h in vitro. Cells were treated with either matrine or control serum for 48 h, followed by extraction of total RNA. The RNA was sequenced using HiSeq 2500 high-throughput sequencing platform (Illumina). A gene library was established and quality analysis of read data carried out. Integrated database from the website DAVID was used to analyze Gene Ontology (GO), and Kyoto encyclopedia of genes and genomes (KEGG) pathway of differential genes was used for pathway analysis, screening for fold differences of more than two times. The FOXO and PI3K-AKT signaling pathways were screened, and expression levels of mRNA and core protein detected by real-time PCR and immunoblotting, respectively. High throughput sequencing and GO analysis revealed that differentially expressed genes before and after treatment played an important role in cell metabolic process, growth process, anatomical structure formation, cellular component organization, and biological regulation. KEGG signal pathway analysis revealed that FOXO and PI3K-AKT signal pathways had a significant difference between before and after matrine-treated androgen-independent prostate cancer cells PC-3 and DU145. Real-time PCR showed that matrine treatment led to a significant increase in the expression levels of FOXO1A, FOXO3A, FOXO4, and FOXO6 in DU145 and PC-3 cells (P<0.01 or P<0.05), whereas the PI3K expression levels decreased (P<0.01). Similarly, immunoblotting revealed a significant increase (P<0.05) in the expression levels of FOXO1A FOXO3A, FOXO4, and FOXO6 in both PC-3 and DU145 cells, whereas PI3K expression levels decreased (P<0.05). Matrine had a broad regulating effect on the mRNA expression profiles of both PC-3 and DU145 cells. Matrine may inhibit cell proliferation, migration, as well as invasion, and induce apoptosis in both PC-3 and DU145 cells through FOXO and PI3K-AKT signaling pathways. Matrine could therefore be used as a complementary drug to present chemotherapeutic agents, for treating androgen-independent prostate cancer.


matrine androgen-independent prostate cancer mRNA FOXO signaling pathway PI3K-AKT signaling pathway 


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This work was supported by the National Natural Science Foundation of China (81472382), the National Natural Science Foundation of China for Young Scientists (81101947), the Guangdong Province Natural Science Foundation (2014A030313079), the Fundamental Research Funds for the Central Universities (14ykpy19), Guangdong Province Science and Technology for Social Development Project (2013B021800107), Guangzhou City in 2015 scientific research projects (7415600066401 to Hai Huang).


  1. Accili, D., and Arden, K.C. (2004). FoxOs at the crossroads of cellular metabolism, differentiation, and transformation. Cell 117, 421–426.CrossRefPubMedGoogle Scholar
  2. Anders, S., and Huber, W. (2010). Differential expression analysis for sequence count data. Genome Biol 11, R106.CrossRefPubMedPubMedCentralGoogle Scholar
  3. Beelen, K., Hoefnagel, L.D.C., Opdam, M., Wesseling, J., Sanders, J., Vincent, A.D., van Diest, P.J., and Linn, S.C. (2014). PI3K/AKT/mTOR pathway activation in primary and corresponding metastatic breast tumors after adjuvant endocrine therapy. Int J Cancer 135, 1257–1263.CrossRefPubMedPubMedCentralGoogle Scholar
  4. Brenkman, A.B., de Keizer, P.L.J., van den Broek, N.J.F., Jochemsen, A.G., and Burgering, B.M.T. (2008). Mdm2 induces mono-ubiquitination of FOXO4. PLoS ONE 3, e2819.CrossRefPubMedPubMedCentralGoogle Scholar
  5. Calnan, D.R., and Brunet, A. (2008). The FoxO code. Oncogene 27, 2276–2288.CrossRefPubMedGoogle Scholar
  6. Chen, W., Zheng, R., Baade, P.D., Zhang, S., Zeng, H., Bray, F., Jemal, A., Yu, X.Q., and He, J. (2016). Cancer statistics in China, 2015. CA Cancer J Clin 66, 115–132.CrossRefPubMedGoogle Scholar
  7. Fei, M., Zhao, Y., Wang, Y., Lu, M., Cheng, C., Huang, X., Zhang, D., Lu, J., He, S., and Shen, A. (2009). Low expression of Foxo3a is associated with poor prognosis in ovarian cancer patients. Cancer Invest 27, 52–59.CrossRefPubMedGoogle Scholar
  8. Guerin, M., Qian, C., Zhong, Q., Cui, Q., Guo, Y., Bei, J., Shao, J., Zhu, X., Huang, W., Wu, J., Liu, R., Liu, Q., Wang, J., Jia, W., Zheng, X., and Zeng, Y. (2016). Translational oncology toward benefiting cancer patients: the Sun Yat-sen University Cancer Center experience. Sci China Life Sci 59, 1057–1062.CrossRefPubMedGoogle Scholar
  9. Hagenbuchner, J., Kuznetsov, A., Hermann, M., Hausott, B., Obexer, P., and Ausserlechner, M.J. (2012). FOXO3-induced reactive oxygen species are regulated by BCL2L11 (Bim) and SESN3. J Cell Sci 125, 1191–1203.CrossRefPubMedGoogle Scholar
  10. Halacli, S.O., and Dogan, A.L. (2015). FOXP1 regulation via the PI3K/Akt/p70S6K signaling pathway in breast cancer cells. Oncol Lett 9, 1482–1488.CrossRefPubMedPubMedCentralGoogle Scholar
  11. Huang, H., and Tindall, D.J. (2007). Dynamic FoxO transcription factors. J Cell Sci 120, 2479–2487.CrossRefPubMedGoogle Scholar
  12. Klotz, L.O., Sánchez-Ramos, C., Prieto-Arroyo, I., Urbánek, P., Steinbrenner, H., and Monsalve, M. (2015). Redox regulation of FoxO transcription factors. Redox Biol 6, 51–72.CrossRefPubMedPubMedCentralGoogle Scholar
  13. Kumar, N., Crocker, T., Smith, T., Pow-Sang, J., Spiess, P.E., Connors, S., Chornukur, G., Dickinson, S.I., Bai, W., Williams, C.R., Salup, R., and Fu, W. (2012). Prostate cancer chemoprevention targeting high risk populations: model for trial design and outcome measures. J Cancer Sci Ther 2011, pii: 007.PubMedPubMedCentralGoogle Scholar
  14. Lai, J.P., He, X.W., Jiang, Y., and Chen, F. (2003). Preparative separation and determination of matrine from the Chinese medicinal plant Sophora flavescens Ait by molecularly imprinted solid-phase extraction. Anal Bioanal Chem 375, 264–269.CrossRefPubMedGoogle Scholar
  15. Li, Q., Lai, Y., Wang, C., Xu, G., He, Z., Shang, X., Sun, Y., Zhang, F., Liu, L., and Huang, H. (2016). Matrine inhibits the proliferation, invasion and migration of castration-resistant prostate cancer cells through regulation of the NF-κB signaling pathway. Oncol Rep 35, 375–381.CrossRefPubMedGoogle Scholar
  16. Liu, J.Y., Hu, J.H., Zhu, Q.G., Li, F.Q., Wang, J., and Sun, H.J. (2007). Effect of matrine on the expression of substance P receptor and inflammatory cytokines production in human skin keratinocytes and fibroblasts. Int Immunopharmacol 7, 816–823.CrossRefPubMedGoogle Scholar
  17. Medema, R.H., Kops, G.J.P.L., Bos, J.L., and Burgering, B.M. (2000). AFXlike Forkhead transcription factors mediate cell-cycle regulation by Ras and PKB through p27kip1. Nature 404, 782–787.CrossRefPubMedGoogle Scholar
  18. Miller, K.D., Siegel, R.L., Lin, C.C., Mariotto, A.B., Kramer, J.L., Rowland, J.H., Stein, K.D., Alteri, R., and Jemal, A. (2016). Cancer treatment and survivorship statistics, 2016. CA Cancer J Clin 66, 271–289.CrossRefPubMedGoogle Scholar
  19. Muranen, T., Selfors, L.M., Worster, D.T., Iwanicki, M.P., Song, L., Morales, F.C., Gao, S., Mills, G.B., and Brugge, J.S. (2012). Inhibition of PI3K/mTOR leads to adaptive resistance in matrix-attached cancer cells. Cancer Cell 21, 227–239.CrossRefPubMedPubMedCentralGoogle Scholar
  20. Newton, R.H., and Turka, L.A. (2012). Regulation of T cell homeostasis and responses by pten. Front Immun 3, 151.CrossRefGoogle Scholar
  21. Plas, D.R., and Thompson, C.B. (2003). Akt Activation promotes degradation of tuberin and FOXO3a via the proteasome. J Biol Chem 278, 12361–12366.CrossRefPubMedGoogle Scholar
  22. Quon, H., and Loblaw, D.A. (2010). Androgen deprivation therapy for prostate cancer-review of indications in 2010. Curr Oncol 17 Suppl 2, S38–S44.PubMedPubMedCentralGoogle Scholar
  23. Salih, D.A.M., and Brunet, A. (2008). FoxO transcription factors in the maintenance of cellular homeostasis during aging. Curr Opin Cell Biol 20, 126–136.CrossRefPubMedPubMedCentralGoogle Scholar
  24. Siegel, R.L., Miller, K.D., and Jemal, A. (2016). Cancer statistics, 2016. CA Cancer J Clin 66, 7–30.CrossRefPubMedGoogle Scholar
  25. Singh, A., Plati, J., and Khosravi-Far, R. (2011). Harnessing the tumor suppressor function of FOXO as an alternative therapeutic approach in cancer. Curr Drug Targets 12, 1311–1321.CrossRefPubMedGoogle Scholar
  26. Trapnell, C., Roberts, A., Goff, L., Pertea, G., Kim, D., Kelley, D.R., Pimentel, H., Salzberg, S.L., Rinn, J.L., and Pachter, L. (2012). Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nat Protoc 7, 562–578.CrossRefPubMedPubMedCentralGoogle Scholar
  27. van der Vos, K.E., and Coffer, P.J. (2011). The extending network of FOXO transcriptional target genes. Antioxid Redox Signal 14, 579–592.CrossRefPubMedGoogle Scholar
  28. Wang, L., Feng, Z., Wang, X., Wang, X., and Zhang, X. (2010). DEGseq: an R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics 26, 136–138.CrossRefPubMedGoogle Scholar
  29. Wang, Y., Zhou, Y., and Graves, D.T. (2014). FOXO transcription factors: their clinical significance and regulation. BioMed Res Int 2014, 1–13.Google Scholar
  30. Wilk, A., Urbanska, K., Grabacka, M., Mullinax, J., Marcinkiewicz, C., Impastato, D., Estrada, J.J., and Reiss, K. (2012). Fenofibrate-induced nuclear translocation of FoxO3A triggers Bim-mediated apoptosis in glioblastoma cells in vitro. Cell Cycle 11, 2660–2671.CrossRefPubMedPubMedCentralGoogle Scholar
  31. Xie, L., Ushmorov, A., Leithäuser, F., Guan, H., Steidl, C., Färbinger, J., Pelzer, C., Vogel, M.J., Maier, H.J., Gascoyne, R.D., Möller, P., and Wirth, T. (2012). FOXO1 is a tumor suppressor in classical Hodgkin lymphoma. Blood 119, 3503–3511.CrossRefPubMedGoogle Scholar
  32. Zeng, C.W., Wang, W.T., Yu, X.B., Yang, L.J., Chen, S.H., and Li, Y.Q. (2015). Pathways related to PMA-differentiated THP1 human monocytic leukemia cells revealed by RNA-Seq. Sci China Life Sci 58, 1282–1287.CrossRefPubMedGoogle Scholar
  33. Zhang, J., Li, Y., Chen, X., Liu, T., Chen, Y., He, W., Zhang, Q., and Liu, S. (2011). Autophagy is involved in anticancer effects of matrine on SGC-7901 human gastric cancer cells. Oncol Rep 26, 115–124.PubMedGoogle Scholar
  34. Zhang, Y., Tseng, C.C., Tsai, Y.L., Fu, X., Schiff, R., and Lee, A.S. (2013). Cancer cells resistant to therapy promote cell surface relocalization of GRP78 which complexes with PI3K and enhances PI(3,4,5)P3 production. PLoS ONE 8, e80071.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Science China Press and Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  • Qi Li
    • 1
    • 2
  • Hai Huang
    • 3
  • Zheng He
    • 4
  • Yi Sun
    • 1
  • Yufeng Tang
    • 2
  • Xiaohong Shang
    • 2
  • Chengbin Wang
    • 1
    Email author
  1. 1.Department of Clinical LaboratoryThe PLA General HospitalBeijingChina
  2. 2.Department of Clinical Laboratory of Xiyuan HospitalChina Academy of Chinese Medical SciencesBeijingChina
  3. 3.Department of Urology, The Sun Yat-sen Memorial HospitalSun Yat-sen UniversityGuangzhouChina
  4. 4.Beijing Center for Physical and Chemical AnalysisBeijingChina

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