Antiproliferative and Antimetastatic Effects of Praeruptorin C on Human Non–Small Cell Lung Cancer through Inactivating ERK/CTSD Signalling Pathways
Abstract
:1. Introduction
2. Results
2.1. Effect of PC on Cell Viability and Cytotoxicity in NSCLC Cells
2.2. Effect of PC on Cell Cycle Arrest in NSCLC Cells
2.3. PC Inhibits Cell Migration, Invasion and CTSD Expression in NSCLC Cells
2.4. PC inhibits the Activation of the ERK1/2 Pathway in NSCLC Cells
2.5. ERK1/2 Activation is Involved in the PC-Induced Suppression of Cell Migration and Invasion and CTSD Expression in NSCLC Cells
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Culture
4.3. Cell Viability Assay
4.4. Colony Formation Assay
4.5. Cell Cycle Distribution by Flow Cytometric Analysis
4.6. Immunoblotting
4.7. Cell Motility Determined by Wound Healing Assay
4.8. Migration and Invasion Assay
4.9. Reverse Transcription and Real-Time PCR Assay
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
- Siegel, R.L.; Miller, K.D.; Jemal, A. Cancer statistics, 2018. CA Cancer J. Clin. 2018, 68, 7–30. [Google Scholar] [CrossRef] [PubMed]
- Bray, F.; Ferlay, J.; Soerjomataram, I.; Siegel, R.L.; Torre, L.A.; Jemal, A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin. 2018, 68, 394–424. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bender, E. Epidemiology: The dominant malignancy. Nature 2014, 513, S2–S3. [Google Scholar] [CrossRef] [PubMed]
- Politi, K.; Herbst, R.S. Lung cancer in the era of precision medicine. Clin. Cancer Res. 2015, 21, 2213–2220. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Riihimaki, M.; Hemminki, A.; Fallah, M.; Thomsen, H.; Sundquist, K.; Sundquist, J.; Hemminki, K. Metastatic sites and survival in lung cancer. Lung Cancer 2014, 86, 78–84. [Google Scholar] [CrossRef]
- Morgensztern, D.; Ng, S.H.; Gao, F.; Govindan, R. Trends in stage distribution for patients with non-small cell lung cancer: A National Cancer Database survey. J. Thorac. Oncol. 2010, 5, 29–33. [Google Scholar] [CrossRef] [Green Version]
- Rao, J.S. Molecular mechanisms of glioma invasiveness: The role of proteases. Nat. Rev. Cancer 2003, 3, 489–501. [Google Scholar] [CrossRef]
- Huang, S.F.; Horng, C.T.; Hsieh, Y.S.; Hsieh, Y.H.; Chu, S.C.; Chen, P.N. Epicatechin-3-gallate reverses TGF-beta1-induced epithelial-to-mesenchymal transition and inhibits cell invasion and protease activities in human lung cancer cells. Food Chem. Toxicol. 2016, 94, 1–10. [Google Scholar] [CrossRef]
- Webb, C.P.; Vande Woude, G.F. Genes that regulate metastasis and angiogenesis. J. Neurooncol. 2000, 50, 71–87. [Google Scholar] [CrossRef]
- Sis, B.; Sagol, O.; Kupelioglu, A.; Sokmen, S.; Terzi, C.; Fuzun, M.; Ozer, E.; Bishop, P. Prognostic significance of matrix metalloproteinase-2, cathepsin D, and tenascin-C expression in colorectal carcinoma. Pathol. Res. Practice 2004, 200, 379–387. [Google Scholar] [CrossRef]
- Huang, Y.; Chu, T.; Liao, T.; Hu, X.; Huang, B. Downregulation of lysosomal and further gene expression characterization in lung cancer patients with bone metastasis. Artif. Cells Nanomed. Biotechnol. 2017, 45, 758–764. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lu, K.H.; Chen, P.N.; Hsieh, Y.H.; Lin, C.Y.; Cheng, F.Y.; Chiu, P.C.; Chu, S.C.; Hsieh, Y.S. 3-Hydroxyflavone inhibits human osteosarcoma U2OS and 143B cells metastasis by affecting EMT and repressing u-PA/MMP-2 via FAK-Src to MEK/ERK and RhoA/MLC2 pathways and reduces 143B tumor growth in vivo. Food Chem. Toxicol. 2016, 97, 177–186. [Google Scholar] [CrossRef] [PubMed]
- Cheng, C.W.; Chen, P.M.; Hsieh, Y.H.; Weng, C.C.; Chang, C.W.; Yao, C.C.; Hu, L.Y.; Wu, P.E.; Shen, C.Y. Foxo3a-mediated overexpression of microRNA-622 suppresses tumor metastasis by repressing hypoxia-inducible factor-1alpha in ERK-responsive lung cancer. Oncotarget 2015, 6, 44222–44238. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pezzani, R.; Salehi, B.; Vitalini, S.; Iriti, M.; Zuniga, F.A.; Sharifi-Rad, J.; Martorell, M.; Martins, N. Synergistic Effects of Plant Derivatives and Conventional Chemotherapeutic Agents: An Update on the Cancer Perspective. Medicina (Kaunas) 2019, 55, 110. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Caesar, L.K.; Cech, N.B. Synergy and antagonism in natural product extracts: When 1 + 1 does not equal 2. Nat. Prod. Rep. 2019, 36, 869–888. [Google Scholar] [CrossRef] [Green Version]
- Zhu, J.J.; Jiang, J.G. Pharmacological and Nutritional Effects of Natural Coumarins and Their Structure-Activity Relationships. Mol. Nutr. Food Res. 2018, 62, e1701073. [Google Scholar] [CrossRef]
- Wenjie, W.; Houqing, L.; Liming, S.; Ping, Z.; Gengyun, S. Effects of praeruptorin C on blood pressure and expression of phospholamban in spontaneously hypertensive rats. Phytomedicine 2014, 21, 195–198. [Google Scholar] [CrossRef]
- Yu, P.J.; Jin, H.; Zhang, J.Y.; Wang, G.F.; Li, J.R.; Zhu, Z.G.; Tian, Y.X.; Wu, S.Y.; Xu, W.; Zhang, J.J.; et al. Pyranocoumarins isolated from Peucedanum praeruptorum Dunn suppress lipopolysaccharide-induced inflammatory response in murine macrophages through inhibition of NF-kappaB and STAT3 activation. Inflammation 2012, 35, 967–977. [Google Scholar] [CrossRef]
- Yang, L.; Li, X.B.; Yang, Q.; Zhang, K.; Zhang, N.; Guo, Y.Y.; Feng, B.; Zhao, M.G.; Wu, Y.M. The neuroprotective effect of praeruptorin C against NMDA-induced apoptosis through down-regulating of GluN2B-containing NMDA receptors. Toxicol. In Vitro 2013, 27, 908–914. [Google Scholar] [CrossRef]
- Losch, A.; Schindl, M.; Kohlberger, P.; Lahodny, J.; Breitenecker, G.; Horvat, R.; Birner, P. Cathepsin D in ovarian cancer: Prognostic value and correlation with p53 expression and microvessel density. Gynecologic Oncol. 2004, 92, 545–552. [Google Scholar] [CrossRef]
- Chai, Y.; Wu, W.; Zhou, C.; Zhou, J. The potential prognostic value of cathepsin D protein in serous ovarian cancer. Arch. Gynecol. Obstetrics 2012, 286, 465–471. [Google Scholar] [CrossRef] [PubMed]
- Winiarski, B.K.; Cope, N.; Alexander, M.; Pilling, L.C.; Warren, S.; Acheson, N.; Gutowski, N.J.; Whatmore, J.L. Clinical Relevance of Increased Endothelial and Mesothelial Expression of Proangiogenic Proteases and VEGFA in the Omentum of Patients with Metastatic Ovarian High-Grade Serous Carcinoma. Translational Oncol. 2014, 7, 267–276. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rochefort, H. Cathepsin D in breast cancer: A tissue marker associated with metastasis. Eur. J. Cancer 1992, 28A, 1780–1783. [Google Scholar] [CrossRef]
- Ferrandina, G.; Scambia, G.; Bardelli, F.; Benedetti Panici, P.; Mancuso, S.; Messori, A. Relationship between cathepsin-D content and disease-free survival in node-negative breast cancer patients: A meta-analysis. Br. J. Cancer 1997, 76, 661–666. [Google Scholar] [CrossRef] [Green Version]
- Foekens, J.A.; Look, M.P.; Bolt-de Vries, J.; Meijer-van Gelder, M.E.; van Putten, W.L.; Klijn, J.G. Cathepsin-D in primary breast cancer: Prognostic evaluation involving 2810 patients. Br. J. Cancer 1999, 79, 300–307. [Google Scholar] [CrossRef] [Green Version]
- Zhang, C.; Zhang, M.; Song, S. Cathepsin D enhances breast cancer invasion and metastasis through promoting hepsin ubiquitin-proteasome degradation. Cancer Lett. 2018, 438, 105–115. [Google Scholar] [CrossRef]
- Vetvicka, V.; Vetvickova, J.; Benes, P. Role of enzymatically inactive procathepsin D in lung cancer. Anticancer Res. 2004, 24, 2739–2743. [Google Scholar] [PubMed]
- Gemoll, T.; Epping, F.; Heinrich, L.; Fritzsche, B.; Roblick, U.J.; Szymczak, S.; Hartwig, S.; Depping, R.; Bruch, H.P.; Thorns, C.; et al. Increased cathepsin D protein expression is a biomarker for osteosarcomas, pulmonary metastases and other bone malignancies. Oncotarget 2015, 6, 16517–16526. [Google Scholar] [CrossRef] [Green Version]
- Dubey, V.; Luqman, S. Cathepsin D as a Promising Target for the Discovery of Novel Anticancer Agents. Curr. Cancer Drug Targets 2017, 17, 404–422. [Google Scholar] [CrossRef]
- Hsu, W.H.; Chiou, H.L.; Lin, C.L.; Kao, S.H.; Lee, H.L.; Liu, C.J.; Hsieh, Y.H. Metastasis-associated protein 2 regulates human hepatocellular carcinoma metastasis progression through modulating p38MAPK/MMP2 pathways. J. Cancer 2019, 10, 6716–6725. [Google Scholar] [CrossRef]
- Huang, C.F.; Yang, S.F.; Chiou, H.L.; Hsu, W.H.; Hsu, J.C.; Liu, C.J.; Hsieh, Y.H. Licochalcone A inhibits the invasive potential of human glioma cells by targeting the MEK/ERK and ADAM9 signaling pathways. Food Funct. 2018, 9, 6196–6204. [Google Scholar] [CrossRef] [PubMed]
- Wu, M.H.; Lin, C.L.; Chiou, H.L.; Yang, S.F.; Lin, C.Y.; Liu, C.J.; Hsieh, Y.H. Praeruptorin A Inhibits Human Cervical Cancer Cell Growth and Invasion by Suppressing MMP-2 Expression and ERK1/2 Signaling. Int. J. Mol. Sci. 2017, 19, 10. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Huang, C.F.; Teng, Y.H.; Lu, F.J.; Hsu, W.H.; Lin, C.L.; Hung, C.C.; Tung, J.N.; Hsieh, Y.H.; Liu, C.J. Beta-mangostin suppresses human hepatocellular carcinoma cell invasion through inhibition of MMP-2 and MMP-9 expression and activating the ERK and JNK pathways. Environ. Toxicol. 2017, 32, 2360–2370. [Google Scholar] [CrossRef] [PubMed]
- Chen, Y.; Wu, H.; Wang, X.; Wang, C.; Gan, L.; Zhu, J.; Tong, J.; Li, Z. Huaier Granule extract inhibit the proliferation and metastasis of lung cancer cells through down-regulation of MTDH, JAK2/STAT3 and MAPK signaling pathways. Biomed. Pharmacother. 2018, 101, 311–321. [Google Scholar] [CrossRef]
- Park, W.H. MAPK inhibitors, particularly the JNK inhibitor, increase cell death effects in H2O2-treated lung cancer cells via increased superoxide anion and glutathione depletion. Oncol. Rep. 2018, 39, 860–870. [Google Scholar] [CrossRef]
- Fu, H.; Gao, H.; Qi, X.; Zhao, L.; Wu, D.; Bai, Y.; Li, H.; Liu, X.; Hu, J.; Shao, S. Aldolase A promotes proliferation and G1/S transition via the EGFR/MAPK pathway in non-small cell lung cancer. Cancer Commun. 2018, 38, 18. [Google Scholar] [CrossRef] [Green Version]
- Zhang, Y.X.; Yuan, J.; Gao, Z.M.; Zhang, Z.G. LncRNA TUC338 promotes invasion of lung cancer by activating MAPK pathway. Eur. Rev. Med. Pharmacol. Sci. 2018, 22, 443–449. [Google Scholar] [CrossRef]
- Wang, K.; Ji, W.; Yu, Y.; Li, Z.; Niu, X.; Xia, W.; Lu, S. FGFR1-ERK1/2-SOX2 axis promotes cell proliferation, epithelial-mesenchymal transition, and metastasis in FGFR1-amplified lung cancer. Oncogene 2018, 37, 5340–5354. [Google Scholar] [CrossRef]
- Ding, C.; Tang, W.; Fan, X.; Wang, X.; Wu, H.; Xu, H.; Xu, W.; Gao, W.; Wu, G. Overexpression of PEAK1 contributes to epithelial-mesenchymal transition and tumor metastasis in lung cancer through modulating ERK1/2 and JAK2 signaling. Cell Death Dis. 2018, 9, 802. [Google Scholar] [CrossRef] [Green Version]
- Xing, S.G.; Zhang, K.J.; Qu, J.H.; Ren, Y.D.; Luan, Q. Propofol induces apoptosis of non-small cell lung cancer cells via ERK1/2-dependent upregulation of PUMA. Eur. Rev. Med. Pharmacol. Sci. 2018, 22, 4341–4349. [Google Scholar] [CrossRef]
- Yu, Q.; Ma, L.; Shen, Y.; Zhai, W.; Zhou, Y. Effect of angular pyranocoumarin isolated from peucedanum praeruptorum on the proliferation and apoptosis of U266 cells. Zhonghua Xue Ye Xue Za Zhi 2015, 36, 937–941. [Google Scholar] [CrossRef] [PubMed]
- Li, X.M.; Jiang, X.J.; Yang, K.; Wang, L.X.; Wen, S.Z.; Wang, F. Prenylated Coumarins from Heracleum stenopterum, Peucedanum praeruptorum, Clausena lansium, and Murraya paniculata. Nat. Prod. Bioprospect. 2016, 6, 233–237. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liang, T.; Yue, W.; Li, Q. Chemopreventive effects of Peucedanum praeruptorum DUNN and its major constituents on SGC7901 gastric cancer cells. Molecules 2010, 15, 8060–8071. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Laurent-Matha, V.; Maruani-Herrmann, S.; Prebois, C.; Beaujouin, M.; Glondu, M.; Noel, A.; Alvarez-Gonzalez, M.L.; Blacher, S.; Coopman, P.; Baghdiguian, S.; et al. Catalytically inactive human cathepsin D triggers fibroblast invasive growth. J. Cell Biol. 2005, 168, 489–499. [Google Scholar] [CrossRef] [PubMed]
- Ohri, S.S.; Vashishta, A.; Proctor, M.; Fusek, M.; Vetvicka, V. Depletion of procathepsin D gene expression by RNA interference: A potential therapeutic target for breast cancer. Cancer Biol. Ther. 2007, 6, 1081–1087. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pranjol, M.Z.I.; Gutowski, N.J.; Hannemann, M.; Whatmore, J.L. Cathepsin D non-proteolytically induces proliferation and migration in human omental microvascular endothelial cells via activation of the ERK1/2 and PI3K/AKT pathways. Biochim. Biophys. Acta Mol. Cell Res. 2018, 1865, 25–33. [Google Scholar] [CrossRef]
Sample Availability: Samples of the compounds are not available from the authors. |
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Liu, C.-M.; Shen, H.-T.; Lin, Y.-A.; Yu, Y.-L.; Chen, Y.-S.; Liu, C.-J.; Hsieh, Y.-H. Antiproliferative and Antimetastatic Effects of Praeruptorin C on Human Non–Small Cell Lung Cancer through Inactivating ERK/CTSD Signalling Pathways. Molecules 2020, 25, 1625. https://doi.org/10.3390/molecules25071625
Liu C-M, Shen H-T, Lin Y-A, Yu Y-L, Chen Y-S, Liu C-J, Hsieh Y-H. Antiproliferative and Antimetastatic Effects of Praeruptorin C on Human Non–Small Cell Lung Cancer through Inactivating ERK/CTSD Signalling Pathways. Molecules. 2020; 25(7):1625. https://doi.org/10.3390/molecules25071625
Chicago/Turabian StyleLiu, Chien-Ming, Huan-Ting Shen, Yi-An Lin, Yung-Luen Yu, Yong-Syuan Chen, Chung-Jung Liu, and Yi-Hsien Hsieh. 2020. "Antiproliferative and Antimetastatic Effects of Praeruptorin C on Human Non–Small Cell Lung Cancer through Inactivating ERK/CTSD Signalling Pathways" Molecules 25, no. 7: 1625. https://doi.org/10.3390/molecules25071625