Proteomic analysis of oridonin-induced apoptosis in multiple myeloma cells

Zhao,Jing; Zhang,Mei; He,Pengcheng; Zhao,Junjie; Chen,Ying; Qi,Jun; Wang,Yuan

doi:10.3892/mmr.2017.6213

Proteomic analysis of oridonin-induced apoptosis in multiple myeloma cells

Authors:
- Jing Zhao
- Mei Zhang
- Pengcheng He
- Junjie Zhao
- Ying Chen
- Jun Qi
- Yuan Wang
View Affiliations

Affiliations: Department of Hematology, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Department of Genetics and Molecular Biology, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Institute of Xi'an Blood Bank, Shaanxi Blood Center, Xi'an, Shaanxi 710061, P.R. China
Published online on: February 17, 2017 https://doi.org/10.3892/mmr.2017.6213
Pages: 1807-1815

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Oridonin is a diterpenoid compound isolated from the medicinal herb Rabdosia rubescens, and has shown marked antitumor effects against different types of cancer. However, the definitive systematic molecular mechanism underlying the antitumor activity of oridonin in multiple myeloma remains to be elucidated. In the present study, cell viability and cytotoxicity were examined to determine the appropriate concentration for proteomic investigation. In addition, cell apoptosis was evaluated using flow cytometry and transmission electron microscopy. A proteomic investigation using a two‑dimensional electrophoresis system and mass spectrometry was performed to identify and characterize the global proteome of the apoptosis induced by oridonin. Of the proteins identiﬁed, seven were involved in the anticancer effects of oridonin. Regulation of the expression and function of target proteins, stathmin, dihydrofolate reductase and pyruvate dehydrogenase E1β, may be potential, therapeutic strategies to effectively treat multiple myeloma. These ﬁndings provide novel information on the molecular mechanisms underlying the anticancer properties of oridonin in multiple myeloma.

View Figures

View References

1	Sirohi B and Powles R: Multiple myeloma. Lancet. 363:875–887. 2004. View Article : Google Scholar : PubMed/NCBI
2	Mahindra A, Laubach J, Raje N, Munshi N, Richardson PG and Anderson K: Latest advances and current challenges in the treatment of multiple myeloma. Nat Rev Clin Oncol. 9:135–143. 2012. View Article : Google Scholar : PubMed/NCBI
3	Cheng Y, Qiu F, Ye YC, Tashiro S, Onodera S and Ikejima T: Oridonin induces G2/M arrest and apoptosis via activating ERK-p53 apoptotic pathway and inhibiting PTK-Ras-Raf-JNK survival pathway in murine fibrosarcoma L929 cells. Arch Biochem Biophys. 490:70–75. 2009. View Article : Google Scholar : PubMed/NCBI
4	Yang J, Jiang H, Wang C, Yang B, Zhao L, Hu D, Qiu G, Dong X and Xiao B: Oridonin triggers apoptosis in colorectal carcinoma cells and suppression of microRNA-32 expression augments oridonin-mediated apoptotic effects. Biomed Pharmacother. 72:125–134. 2015. View Article : Google Scholar : PubMed/NCBI
5	Kang N, Zhang JH, Qiu F, Tashiro S, Onodera S and Ikejima T: Inhibition of EGFR signaling augments oridonin-induced apoptosis in human laryngeal cancer cells via enhancing oxidative stress coincident with activation of both the intrinsic and extrinsic apoptotic pathways. Cancer Lett. 294:147–158. 2010. View Article : Google Scholar : PubMed/NCBI
6	Lou H, Zhang X, Gao L, Feng F, Wang J, Wei X, Yu Z, Zhang D and Zhang Q: In vitro and in vivo antitumor activity of oridonin nanosuspension. Int J Pharm. 379:181–186. 2009. View Article : Google Scholar : PubMed/NCBI
7	Liu YQ, Mu ZQ, You S, Tashiro S, Onodera S and Ikejima T: Fas/FasL Signaling allows extracelluar-signal regulated kinase to regulate cytochrome c release in oridonin-induced apoptotic u937 cells. Biol Pharm Bull. 29:1873–1879. 2006. View Article : Google Scholar : PubMed/NCBI
8	Ren KK, Wang HZ, Xie LP, Chen DW, Liu X, Sun J, Nie YC and Zhang RQ: The effects of oridonin on cell growth, cell cycle, cell migration and differentiation in melanoma cells. J Ethnopharmacol. 103:176–180. 2006. View Article : Google Scholar : PubMed/NCBI
9	Zhang Y, Wu Y, Tashiro S, Onodera S and Ikejima T: Involvement of PKC signal pathways in oridonin-induced autophagy in HeLa cells: A protective mechanism against apoptosis. Biochem Biophys Res Commun. 378:273–278. 2009. View Article : Google Scholar : PubMed/NCBI
10	Hsieh TC, Wijeratne EK, Liang JY, Gunatilaka AL and Wu JM: Differential control of growth, cell cycle progression and expression of NF-kappaB in human breast cancer cells MCF-7, MCF-10A, and MDA-MB-231 by ponicidin and oridonin, diterpenoids from the chinese herb Rabdosia rubescens. Biochem Biophys Res Commun. 337:224–231. 2005. View Article : Google Scholar : PubMed/NCBI
11	Hanash SM, Madoz-Gurpide J and Misek DE: Identification of novel targets for cancer therapy using expression proteomics. Leukemia. 16:478–485. 2002. View Article : Google Scholar : PubMed/NCBI
12	Ramagli L: Quantifying protein in 2-D PAGE solubilization buffers2-D Proteome Analysis Protocols. Link AJ: 112. Humana Press; Totowa, NJ: pp. 99–103. 1999, View Article : Google Scholar
13	Görg A, Postel W and Günther S: The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis. 9:531–546. 1988. View Article : Google Scholar : PubMed/NCBI
14	B Tom ST: 2-D electrophoresis using immobilized pH gradients, pricinples and methods. Amersham Pharmacia Biotech. 1998.
15	Pasquali C, Fialka I and Huber LA: Preparative two-dimensional gel electrophoresis of membrane proteins. Electrophosis. 18:2573–2781. 1997. View Article : Google Scholar
16	Iversen LF, Kastrup JS, Bjørn SE, Wiberg FC, Larsen IK, Flodgaard HJ and Rasmussen PB: Structure and function of the N-linked glycans of HBP/CAP37/azurocidin: Crystal structure determination and biological characterization of nonglycosylated HBP. Protein Sci. 8:2019–2026. 1999. 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	Zhu Y, Xie L, Chen G, Chen G, Wang H and Zhang R: Effects of oridonin on proliferation of HT29 human colon carcinoma cell lines both in vitro and in vivo in mice. Pharmazie. 62:439–444. 2007.PubMed/NCBI
19	Dal Piaz F, Cotugno R, Lepore L, Vassallo A, Malafronte N, Lauro G, Bifulco G, Belisario MA and De Tommasi N: Chemical proteomics reveals HSP70 1A as a target for the anticancer diterpene oridonin in Jurkat cells. J Proteomics. 82:14–26. 2013. View Article : Google Scholar : PubMed/NCBI
20	Charbaut E, Curmi PA, Ozon S, Lachkar S, Redeker V and Sobel A: Stathmin family proteins display specific molecular and tubulin binding properties. J Biol Chem. 276:16146–16154. 2001. View Article : Google Scholar : PubMed/NCBI
21	Alli E, Yang JM and Hait WN: Silencing of stathmin induces tumor-suppressor function in breast cancer cell lines harboring mutant p53. Oncogene. 26:1003–1012. 2007. View Article : Google Scholar : PubMed/NCBI
22	Mistry SJ, Bank A and Atweh GF: Targeting stathmin in prostate cancer. Mol Cancer Ther. 4:1821–1829. 2005. View Article : Google Scholar : PubMed/NCBI
23	Mistry SJ and Atweh GF: Therapeutic interactions between stathmin inhibition and chemotherapeutic agents in prostate cancer. Mol Cancer Ther. 5:3248–3257. 2006. View Article : Google Scholar : PubMed/NCBI
24	Yuan RH, Jeng YM, Chen HL, Lai PL, Pan HW, Hsieh FJ, Lin CY, Lee PH and Hsu HC: Stathmin overexpression cooperates with p53 mutation and osteopontin overexpression, and is associated with tumour progression, early recurrence, and poor prognosis in hepatocellular carcinoma. J Pathol. 209:549–558. 2006. View Article : Google Scholar : PubMed/NCBI
25	Zhang HZ, Wang Y, Gao P, Lin F, Liu L, Yu B, Ren JH, Zhao H and Wang R: Silencing stathmin gene expression by survivin promoter-driven siRNA vector to reverse malignant phenotype of tumor cells. Cancer Biol Ther. 5:1457–1461. 2006. View Article : Google Scholar : PubMed/NCBI
26	Iancu C, Mistry SJ, Arkin S and Atweh GF: Taxol and anti-stathmin therapy: A synergistic combination that targets the mitotic spindle. Cancer Res. 60:3537–3541. 2000.PubMed/NCBI
27	Jensen DE, Black AR, Swick AG and Azizkhan JC: Distinct roles for Sp1 and E2F sites in the growth/cell cycle regulation of the DHFR promoter. J Cell Biochem. 67:24–31. 1997. View Article : Google Scholar : PubMed/NCBI
28	Chen MJ, Shimada T, Moulton AD, Cline A, Humphries RK, Maizel J and Nienhuis AW: The functional human dihydrofolate reductase gene. J Biol Chem. 259:3933–3943. 1984.PubMed/NCBI
29	Assaraf YG: Molecular basis of antifolate resistance. Cancer Metastasis Rev. 26:153–181. 2007. View Article : Google Scholar : PubMed/NCBI
30	Morales C, García MJ, Ribas M, Miró R, Muñoz M, Caldas C and Peinado MA: Dihydrofolate reductase amplification and sensitization to methotrexate of methotrexate-resistant colon cancer cells. Mol Cancer Ther. 8:424–432. 2009. View Article : Google Scholar : PubMed/NCBI
31	Biaglow JE and Miller RA: The thioredoxin reductase/thioredoxin system: Novel redox targets for cancer therapy. Cancer Biol Ther. 4:6–13. 2005. View Article : Google Scholar : PubMed/NCBI
32	Nguyen P, Awwad RT, Smart DD, Spitz DR and Gius D: Thioredoxin reductase as a novel molecular target for cancer therapy. Cancer Lett. 236:164–174. 2006. View Article : Google Scholar : PubMed/NCBI
33	Yoo MH, Xu XM, Carlson BA, Gladyshev VN and Hatfield DL: Thioredoxin reductase 1 deficiency reverses tumor phenotype and tumorigenicity of lung carcinoma cells. J Biol Chem. 281:13005–13008. 2006. View Article : Google Scholar : PubMed/NCBI
34	Lu J, Chew EH and Holmgren A: Targeting thioredoxin reductase is a basis for cancer therapy by arsenic trioxide. Proc Natl Acad Sci USA. 104:12288–12293. 2007. View Article : Google Scholar : PubMed/NCBI
35	Wang X, Zhang J and Xu T: Cyclophosphamide as a potent inhibitor of tumor thioredoxin reductase in vivo. Toxicol Appl Pharmacol. 218:88–95. 2007. View Article : Google Scholar : PubMed/NCBI
36	Glushakova LG, Lisankie MJ, Eruslanov EB, Ojano-Dirain C, Zolotukhin I, Liu C, Srivastava A and Stacpoole PW: AAV3-mediated transfer and expression of the pyruvate dehydrogenase E1 alpha subunit gene causes metabolic remodeling and apoptosis of human liver cancer cells. Mol Genet Metab. 98:289–299. 2009. View Article : Google Scholar : PubMed/NCBI
37	Lu J, Tan M and Cai Q: The Warburg effect in tumor progression: Mitochondrial oxidative metabolism as an anti-metastasis mechanism. Cancer Lett. 356:156–164. 2015. View Article : Google Scholar : PubMed/NCBI
38	Michelakis ED, Webster L and Mackey JR: Dichloroacetate (DCA) as a potential metabolic-targeting therapy for cancer. Br J Cancer. 99:989–994. 2008. View Article : Google Scholar : PubMed/NCBI