Open Access

Pseudolaric acid B induced autophagy, but not apoptosis, in MRC5 human fibroblast cells

  • Authors:
    • Yue Wang
    • Hongmei Gao
    • Ting Wu
    • Zengyan Wang
    • Fengmei Song
    • Antian Chen
    • Jing Zhang
    • Wenyan Zhang
    • Hui Zhang
    • Jinghua Yu
  • View Affiliations

  • Published online on: November 15, 2017     https://doi.org/10.3892/ol.2017.7426
  • Pages: 863-870
  • Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Autophagy and apoptosis are closely associated. In previous studies, pseudolaric acid B (PAB), a diterpene acid isolated from the root and trunk bark of Pseudolarix kaempferi Gordon (Pinaceae), was demonstrated to induce apoptosis in various cell lines. However, in L929 murine fibrosarcoma and SW579 human thyroid squamous cell carcinoma cells, only autophagy was induced. In the present study, another cell line, MRC5 human lung fibroblast cells, was identified in which PAB only induced autophagy. The relationship between apoptosis and autophagy subsequent to PAB treatment in MRC5 cells was explored. When autophagy was inhibited by 3‑methyladenine (3MA), apoptosis was induced in the PAB‑treated MRC5 cells. To study the mechanism for the promotion of apoptosis by 3MA in the PAB‑treated cells, the expression of members from the apoptotic signal pathways was assessed. As Bcl‑2, Bcl‑2 associated X and pro‑caspase‑9 expression following PAB treatment was not affected by 3MA treatment, it was determined that apoptosis was induced independent of the mitochondrial pathway of apoptosis. As Fas and pro‑caspase‑8 expression following PAB treatment were not altered by 3MA, it was further determined that the death receptor pathway was not induced. However, the phosphorylation of c‑Jun‑N‑terminal kinase and the expression of pro‑caspase‑3 were upregulated, and the phosphorylation of extracellular signal‑regulated kinase downregulated, by the combination of PAB and 3MA treatment compared with PAB alone. It was also observed that 3MA did not affect the microtubule aggregation ability of PAB. Therefore, inhibiting autophagy in MRC5 cells did not affect the role of PAB in microtubule aggregation, while apoptosis was induced. This may present a strategy to enhance the anti‑tumor effects of PAB.

References

1 

Gong X, Wang M, Tashiro S, Onodera S and Ikejima T: Involvement of JNK-initiated p53 accumulation and phosphorylation of p53 in pseudolaric acid B induced cell death. Exp Mol Med. 38:428–434. 2006. View Article : Google Scholar : PubMed/NCBI

2 

Gong XF, Wang MW, Tashiro S, Onodera S and Ikejima T: Pseudolaric acid B induces apoptosis through p53 and Bax/Bcl-2 pathways in human melanoma A375-S2 cells. Arch Pharm Res. 28:68–72. 2005. View Article : Google Scholar : PubMed/NCBI

3 

Pan DJ, Li ZL, Hu CQ, Chen K, Chang JJ and Lee KH: The cytotoxic principles of Pseudolarix kaempferi: Pseudolaric acid-A and -B and related derivatives. Planta Med. 56:383–385. 1990. View Article : Google Scholar : PubMed/NCBI

4 

Yu J, Li X, Tashiro S, Onodera S and Ikejima T: Bcl-2 family proteins were involved in pseudolaric acid B-induced autophagy in murine fibrosarcoma L929 cells. J Pharmacol Sci. 107:295–302. 2008. View Article : Google Scholar : PubMed/NCBI

5 

Yu JH, Cui Q, Jiang YY, Yang W, Tashiro S, Onodera S and Ikejima T: Pseudolaric acid B induces apoptosis, senescence, and mitotic arrest in human breast cancer MCF-7. Acta Pharmacol Sin. 28:1975–1983. 2007. View Article : Google Scholar : PubMed/NCBI

6 

Yu JH, Wang HJ, Li XR, Tashiro S, Onodera S and Ikejima T: Protein tyrosine kinase, JNK, and ERK involvement in pseudolaric acid B-induced apoptosis of human breast cancer MCF-7 cells. Acta Pharmacol Sin. 29:1069–1076. 2008. View Article : Google Scholar : PubMed/NCBI

7 

Li FF, Yi S, Wen L, He J, Yang LJ, Zhao J, Zhang BP, Cui GH and Chen Y: Oridonin induces NPM mutant protein translocation and apoptosis in NPM1c+ acute myeloid leukemia cells in vitro. Acta Pharmacol Sin. 35:806–813. 2014. View Article : Google Scholar : PubMed/NCBI

8 

Qi M, Yao G, Fan S, Cheng W, Tashiro S, Onodera S and Ikejima T: Pseudolaric acid B induces mitotic catastrophe followed by apoptotic cell death in murine fibrosarcoma L929 cells. Eur J Pharmacol. 683:16–26. 2012. View Article : Google Scholar : PubMed/NCBI

9 

Adams JM: Ways of dying: Multiple pathways to apoptosis. Genes Dev. 17:2481–2495. 2003. View Article : Google Scholar : PubMed/NCBI

10 

Nagata S and Golstein P: The Fas death factor. Science. 267:1449–1456. 1995. View Article : Google Scholar : PubMed/NCBI

11 

Klekotka PA, Santoro SA, Wang H and Zutter MM: Specific residues within the alpha 2 integrin subunit cytoplasmic domain regulate migration and cell cycle progression via distinct MAPK pathways. J Biol Chem. 276:32353–32361. 2001. View Article : Google Scholar : PubMed/NCBI

12 

Xia Z, Dickens M, Raingeaud J, Davis RJ and Greenberg ME: Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science. 270:1326–1331. 1995. View Article : Google Scholar : PubMed/NCBI

13 

Yoon S and Seger R: The extracellular signal-regulated kinase: Multiple substrates regulate diverse cellular functions. Growth Factors. 24:21–44. 2006. View Article : Google Scholar : PubMed/NCBI

14 

Lee YJ, Won AJ, Lee J, Jung JH, Yoon S, Lee BM and Kim HS: Molecular mechanism of SAHA on regulation of autophagic cell death in tamoxifen-resistant MCF-7 breast cancer cells. Int J Med Sci. 9:881–893. 2012. View Article : Google Scholar : PubMed/NCBI

15 

Lee YZ, Yang CW, Chang HY, Hsu HY, Chen IS, Chang HS, Lee CH, Lee JC, Kumar CR, Qiu YQ, et al: Discovery of selective inhibitors of Glutaminase-2, which inhibit mTORC1, activate autophagy and inhibit proliferation in cancer cells. Oncotarget. 5:6087–6101. 2014. View Article : Google Scholar : PubMed/NCBI

16 

He H, Feng YS, Zang LH, Liu WW, Ding LQ, Chen LX, Kang N, Hayashi T, Tashiro S, Onodera S, et al: Nitric oxide induces apoptosis and autophagy; autophagy down-regulates NO synthesis in physalin A-treated A375-S2 human melanoma cells. Food Chem Toxicol. 71:128–135. 2014. View Article : Google Scholar : PubMed/NCBI

17 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI

18 

Wang ZY, Zhong T, Wang Y, Song FM and Yu XF, Xing LP, Zhang WY, Yu JH, Hua SC and Yu XF: Human enterovirus 68 interferes with the host cell cycle to facilitate viral production. Front Cell Infect Microbiol. 7:292017. View Article : Google Scholar : PubMed/NCBI

19 

Zhong T, Zhang LY, Wang ZY, Wang Y, Song FM, Zhang YH and Yu JH: Rheum emodin inhibits enterovirus 71 viral replication and affects the host cell cycle environment. Acta Pharmacol Sin. 38:392–401. 2017. View Article : Google Scholar : PubMed/NCBI

20 

Liu YQ, Han XF, Bo JX and Ma HP: Wedelolactone enhances osteoblastogenesis but inhibits osteoclastogenesis through Sema3A/NRP1/PlexinA1 pathway. Front Pharmacol. 7:3752016. View Article : Google Scholar : PubMed/NCBI

21 

Yu J, Chen C, Xu T, Yan M, Xue B, Wang Y, Liu C, Zhong T, Wang Z, Meng X, et al: Pseudolaric acid B activates autophagy in MCF-7 human breast cancer cells to prevent cell death. Oncol Lett. 11:1731–1737. 2016.PubMed/NCBI

22 

Yu J, Ren P, Zhong T, Wang Y, Yan M, Xue B, Li R, Dai C, Liu C, Chen G and Yu XF: Pseudolaric acid B inhibits proliferation in SW579 human thyroid squamous cell carcinoma. Mol Med Rep. 12:7195–7202. 2015. View Article : Google Scholar : PubMed/NCBI

23 

Yu J, Wang Z, Ren P, Zhong T, Wang Y, Song F, Hou J, Yu X and Hua S: Pseudolaric acid B inhibits the secretion of hepatitis B virus. Oncol Rep. 37:519–525. 2017. View Article : Google Scholar : PubMed/NCBI

24 

Wong VK, Chiu P, Chung SS, Chow LM, Zhao YZ, Yang BB and Ko BC: Pseudolaric acid B, a novel microtubule-destabilizing agent that circumvents multidrug resistance phenotype and exhibits antitumor activity in vivo. Clin Cancer Res. 11:6002–6011. 2005. View Article : Google Scholar : PubMed/NCBI

25 

Feng Y, He D, Yao Z and Klionsky DJ: The machinery of macroautophagy. Cell Res. 24:24–41. 2014. View Article : Google Scholar : PubMed/NCBI

26 

Galluzzi L, Pietrocola F, Bravo-San Pedro JM, Amaravadi RK, Baehrecke EH, Cecconi F, Codogno P, Debnath J, Gewirtz DA, Karantza V, et al: Autophagy in malignant transformation and cancer progression. EMBO J. 34:856–880. 2015. View Article : Google Scholar : PubMed/NCBI

27 

Crighton D, Wilkinson S, O'Prey J, Syed N, Smith P, Harrison PR, Gasco M, Garrone O, Crook T and Ryan KM: DRAM, a p53-induced modulator of autophagy, is critical for apoptosis. Cell. 126:121–134. 2006. View Article : Google Scholar : PubMed/NCBI

28 

Behrends C, Sowa ME, Gygi SP and Harper JW: Network organization of the human autophagy system. Nature. 466:68–76. 2010. View Article : Google Scholar : PubMed/NCBI

29 

Mariño G, Niso-Santano M, Baehrecke EH and Kroemer G: Self-consumption: The interplay of autophagy and apoptosis. Nat Rev Mol Cell Biol. 15:81–94. 2014. View Article : Google Scholar : PubMed/NCBI

30 

Salah FS, Ebbinghaus M, Muley VY, Zhou Z, Al-Saadi KR, Pacyna-Gengelbach M, O'Sullivan GA, Betz H, König R, Wang ZQ, et al: Tumor suppression in mice lacking GABARAP, an Atg8/LC3 family member implicated in autophagy, is associated with alterations in cytokine secretion and cell death. Cell Death Dis. 7:e22052016. View Article : Google Scholar : PubMed/NCBI

31 

Tong YG, Zhang XW, Geng MY, Yue JM, Xin XL, Tian F, Shen X, Tong LJ, Li MH, Zhang C, et al: Pseudolarix acid B, a new tubulin-binding agent, inhibits angiogenesis by interacting with a novel binding site on tubulin. Mol Pharmacol. 69:1226–1233. 2006. View Article : Google Scholar : PubMed/NCBI

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January 2018
Volume 15 Issue 1

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Copy and paste a formatted citation
APA
Wang, Y., Gao, H., Wu, T., Wang, Z., Song, F., Chen, A. ... Yu, J. (2018). Pseudolaric acid B induced autophagy, but not apoptosis, in MRC5 human fibroblast cells. Oncology Letters, 15, 863-870. https://doi.org/10.3892/ol.2017.7426
MLA
Wang, Y., Gao, H., Wu, T., Wang, Z., Song, F., Chen, A., Zhang, J., Zhang, W., Zhang, H., Yu, J."Pseudolaric acid B induced autophagy, but not apoptosis, in MRC5 human fibroblast cells". Oncology Letters 15.1 (2018): 863-870.
Chicago
Wang, Y., Gao, H., Wu, T., Wang, Z., Song, F., Chen, A., Zhang, J., Zhang, W., Zhang, H., Yu, J."Pseudolaric acid B induced autophagy, but not apoptosis, in MRC5 human fibroblast cells". Oncology Letters 15, no. 1 (2018): 863-870. https://doi.org/10.3892/ol.2017.7426