Tumor Biology

, Volume 37, Issue 1, pp 331–339 | Cite as

Imperatorin acts as a cisplatin sensitizer via downregulating Mcl-1 expression in HCC chemotherapy

  • Jianjian Hu
  • Changlong Xu
  • Bihuan Cheng
  • Lingxiang Jin
  • Jie Li
  • Yuqiang Gong
  • Wei Lin
  • Zhenzhen Pan
  • Chenwei Pan
Research Article

Abstract

Acquisition of cisplatin resistance is the common and critical limitation for hepatocellular carcinoma (HCC) therapy. Our study was aimed to determine whether there were conditions under which the addition of imperatorin would reverse the resistance of HCC cells to cisplatin-based therapy. In this study, we found that addition of imperatorin significantly enhanced the cytotoxicity of cisplatin to HCC cells. Since the Mcl-1 was overexpressed in HCC cell lines (HepG2, HepG3B, PLC, Huh7) compared with normal liver cell line (L-O2), we found that the Mcl-1 expression was downregulated by imperatorin but not influenced by cisplatin in HCC cells. In addition, our results showed the combination of imperatorin and cisplatin induced apoptosis and ∆Ψm collapse more significantly compared with treatment of imperatorin or cisplatin alone. Furthermore, the imperatorin-induced sensitization for cisplatin-cytotoxicity to HCC cells was abolished by the transfection of Mcl-1 expression plasmid. Finally, we found that the addition of imperatorin significantly reversed the resistance to cisplatin in cisplatin-resistant HCC cells, which was Mcl-1 dependent. In summary, our study revealed that combination with imperatorin could enhance the antitumor activity of cisplatin via targeting Mcl-1 and reverse the resistance to cisplatin in HCC.

Keywords

HCC Imperatorin Cisplatin Mcl-1 Apoptosis 

Notes

Acknowledgments

This work was supported by the Program of WenZhou Science and Technology Bureau (grant no. Y20140092)

Conflict of interest

None

Author contributions

CP designed the study. JH, CX, and BC wrote the manuscript. JH, LJ, and JL performed out the immunohistochemistry and the related statistical analysis. JH, YG, WL, and ZP carried out the cell culture and transfection, Western blot, RT-PCR, and flow cytometry. All authors approved the final version of the manuscript.

Supplementary material

13277_2015_3591_MOESM1_ESM.tif (931 kb)
Fig. Supplement The transfection efficiency of pcDNA3.1-Mcl-1 in HCC cells. (A) HepG2 cells and Huh7 cells were transfected with pcDNA3.1-Mcl-1 plasmid or its control. After 24 h incubation, the Mcl-1 mRNA level was detected by quantitative PCR. (B) Mcl-1 protein level was detected by western blot assay after the HepG2 and Huh7 cells were treated as above. (TIFF 931 kb)
13277_2015_3591_Fig6_ESM.gif (706 kb)
High Resolution (GIF 706 kb)

References

  1. 1.
    Li W, Xie L, He X, Li J, Tu K, Wei L, et al. Diagnostic and prognostic implications of microRNAs in human hepatocellular carcinoma. Int J Cancer. 2008;123:1616–22.CrossRefPubMedGoogle Scholar
  2. 2.
    Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90.CrossRefPubMedGoogle Scholar
  3. 3.
    Kerr SH, Kerr DJ. Novel treatments for hepatocellular cancer. Cancer Lett. 2009;286:114–20.CrossRefPubMedGoogle Scholar
  4. 4.
    Hubaux R, Thu KL, Vucic EA, Pikor LA, Kung SH, Martinez VD, et al. Microtubule affinity-regulating kinase 2 is associated with DNA damage response and cisplatin resistance in non-small cell lung cancer. Int J Cancer. 2015.Google Scholar
  5. 5.
    Galluzzi L, Vitale I, Michels J, Brenner C, Szabadkai G, Harel-Bellan A, et al. Systems biology ofcisplatin resistance: past, present and future. Cell Death Dis. 2014;5:e1257.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    O’Grady S, Finn SP, Cuffe S, Richard DJ, O’Byrne KJ, Barr MP. The role of DNA repair pathways in cisplatin resistant lung cancer. Cancer Treat Rev. 2014;40:1161–70.CrossRefPubMedGoogle Scholar
  7. 7.
    Rytelewski M, Tong JG, Koropatnick J, et al. BRCA2 inhibition enhances cisplatin-mediated alterations in tumor cell proliferation, metabolism, and metastasis. Mol Oncol. 2014;8:1429–40.CrossRefPubMedGoogle Scholar
  8. 8.
    Rahman A, Siddiqui SA, Jakhar R, Kang SC. Growth inhibition of various human cancer cell lines by imperatorin and limonin from poncirus trifoliata rafin. Seeds Anticancer Agents Med Chem. 2015;15:236–41.CrossRefPubMedGoogle Scholar
  9. 9.
    García-Argáez AN, Ramírez Apan TO, Parra Delgado H, Velázquez G, Martínez-Vázquez M. Anti-inflammatory activity of coumarins from Decatropis bicolor on TPA ear mice model. Planta Med. 2000;66:279–81.CrossRefPubMedGoogle Scholar
  10. 10.
    Zhang Y, Cao Y, Zhan Y, Duan H, He L. Furanocoumarins-imperatorin inhibits myocardial hypertrophy both in vitro and in vivo. Fitoterapia. 2010;81:1188–95.CrossRefPubMedGoogle Scholar
  11. 11.
    Choochuay K, Chunhacha P, Pongrakhananon V, Luechapudiporn R, Chanvorachote P. Imperatorin sensitizes anoikis and inhibits anchorage-independent growth of lung cancer cells. J Nat Med. 2013;67:599–606.CrossRefPubMedGoogle Scholar
  12. 12.
    Luo KW, Sun JG, Chan JY, Yang L, Wu SH, Fung KP, et al. Anticancer effects of imperatorin isolated from Angelica dahurica: induction of apoptosis in HepG2 cells through both death-receptor- and mitochondria-mediated pathways. Chemotherapy. 2011;57:449–59.CrossRefPubMedGoogle Scholar
  13. 13.
    Bądziul D, Jakubowicz-Gil J, Paduch R, Głowniak K, Gawron A. Combined treatment with quercetin and imperatorin as a potent strategy for killing HeLa and Hep-2 cells. Mol Cell Biochem. 2014;392:213–27.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 2011;25:402–8.CrossRefGoogle Scholar
  15. 15.
    Kelly PN, Strasser A. The role of Bcl-2 and its pro-survival relatives in tumourigenesis and cancer therapy. Cell Death Differ. 2011;18:1414–24.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Varadarajan S, Butterworth M, Wei J, Pellecchia M, Dinsdale D, Cohen GM. Sabutoclax (BI97C1) and BI112D1, putative inhibitors of MCL-1, induce mitochondrial fragmentation either upstream of or independent of apoptosis. Neoplasia. 2013;15:568–78.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Zhang Z, Zheng L, Zhao Z, Shi J, Wang X, Huang J. Grape seed proanthocyanidins inhibit H2O2-induced osteoblastic MC3T3-E1 cell apoptosis via ameliorating H2O2-induced mitochondrial dysfunction. J Toxicol Sci. 2014;39:803–13.CrossRefPubMedGoogle Scholar
  18. 18.
    Prathapan A, Vineetha VP, Raghu KG. Protective effect of Boerhaavia diffusa L. against mitochondrial dysfunction in angiotensin II induced hypertrophy in H9c2 cardiomyoblast cells. PLoS One. 2014;9, e96220.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Colombo F, Trombetta E, Cetrangolo P, Maggioni M, Razini P, De Santis F, et al. Giant lysosomes as a chemotherapy resistance mechanism in hepatocellular carcinoma cells. PLoS One. 2014;9:e114787.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Wang B, Ni Z, Dai X, Qin L, Li X, Xu L, et al. The Bcl-2/xL inhibitor ABT-263 increases the stability of Mcl-1 mRNA and protein in hepatocellular carcinoma cells. Mol Cancer. 2014;13:98.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Wang B, Chen L, Ni Z, Dai X, Qin L, Wu Y, et al. Hsp90 inhibitor 17-AAG sensitizes Bcl-2 inhibitor (−)-gossypol by suppressing ERK-mediated protective autophagy and Mcl-1 accumulation in hepatocellular carcinoma cells. Exp Cell Res. 2014;328:379–87.CrossRefPubMedGoogle Scholar
  22. 22.
    Leuenroth SJ, Grutkoski PS, Ayala A, Simms HH. The loss of Mcl-1 expression in human polymorphonuclear leukocytes promotes apoptosis. J Leukoc Biol. 2000;68:158–66.PubMedGoogle Scholar
  23. 23.
    Thomas L, Lam C, Edwards S. Mcl-1 the molecular regulation of protein function. FEBS Lett. 2010;584:2981–9.CrossRefPubMedGoogle Scholar
  24. 24.
    Sieghart W, Losert D, Wacheck V, et al. Mcl-1 overexpression in hepatocellular carcinoma: a potential target for antisense therapy. J Hepatol. 2006;44:151–7.CrossRefPubMedGoogle Scholar
  25. 25.
    Sieghart W, Losert D, Wacheck V, et al. Mcl-1 overexpression in hepatocellular carcinoma: a potential target for antisense therapy. J Hepatol. 2006;44:151–7.CrossRefPubMedGoogle Scholar
  26. 26.
    Williams MM, Cook RS. Bcl-2 family proteins in breast development and cancer: could Mcl-1 targeting overcome therapeutic resistance? Oncotarget. 2015;6:3519–30.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Wei S, Dong K, Lin F, Wang X, Li B, Shen J, et al. Inducing apoptosis and enhancing chemosensitivity to gemcitabine via RNA interference targeting mcl-1 gene in pancreatic carcinoma cell. Cancer Chemother Pharmacol. 2008;62:1055–64.CrossRefPubMedGoogle Scholar
  28. 28.
    Zhang R, Li Y, Dong X, Peng L, Nie X. MiR-363 sensitizes cisplatin-induced apoptosis targeting in Mcl-1 in breast cancer. Med Oncol. 2014;31:347.CrossRefPubMedGoogle Scholar
  29. 29.
    Pae HO, Oh H, Yun YG, Oh GS, Jang SI, Hwang KM, et al. Imperatorin, a furanocoumarin from Angelica dahurica (Umbelliferae), induces cytochrome c-dependent apoptosis in human promyelocytic leukaemia, HL-60 cells. Pharmacol Toxicol. 2002;91:40–8.CrossRefPubMedGoogle Scholar
  30. 30.
    Bądziul D, Jakubowicz-Gil J, Langner E, Rzeski W, Głowniak K, Gawron A. The effect of quercetin and imperatorin on programmed cell death induction in T98G cells in vitro. Pharmacol Rep. 2014;66:292–300.CrossRefPubMedGoogle Scholar
  31. 31.
    Ziko L, Riad S, Amer M, Zdero R, Bougherara H, Amleh A. Mechanical stress promotes Cisplatin-induced hepatocellular carcinoma cell death. Biomed Res Int. 2015;2015:430569.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Xie X, Hu Y, Xu L, Fu Y, Tu J, Zhao H, et al. The role of miR-125b-mitochondria-caspase-3 pathway in doxorubicin resistance and therapy in human breast cancer. Tumour Biol. 2015.Google Scholar
  33. 33.
    Zhao J, Li X, Zou M, He J, Han Y, Wu D, et al. miR-135a inhibition protects A549 cells from LPS-induced apoptosis by targeting Bcl-2. Biochem Biophys Res Commun. 2014;452:951–7.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Jianjian Hu
    • 1
  • Changlong Xu
    • 2
  • Bihuan Cheng
    • 1
  • Lingxiang Jin
    • 3
  • Jie Li
    • 3
  • Yuqiang Gong
    • 1
  • Wei Lin
    • 3
  • Zhenzhen Pan
    • 4
  • Chenwei Pan
    • 3
  1. 1.Department of Anesthesiology, Critical Care and Pain MedicineThe Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical UniversityWenzhouChina
  2. 2.Department of GastroenterologyThe Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical UniversityWenzhou CityChina
  3. 3.Department of Infectious DiseaseThe Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical UniversityWenzhou CityChina
  4. 4.Department of Infectious DiseaseThe First Affiliated Hospital of Wenzhou Medical UniversityWenzhouChina

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