Abstract
Aim:
To investigate the effects of small interfering RNA of cyclooxygenase-2 (COX-2) on the proliferation and apoptosis of human multiple myeloma RPMI8226 cells and its relation with the Bcl-2 family in vitro.
Methods:
Transcription and expression of COX-2 in human myeloma RPMI8226 cells were checked by RT-PCR and Western blot analysis, respectively. The COX-2 siRNA fragment targeting exon 5 of COX-2 gene was transfected into the cells with the Amaxa nucleofection technique. The inhibition of cell growth was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay. Apoptosis was estimated by Annexin-V/ propidium iodide double-labeled cytometry and confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay. Bcl-2 and Bax expression was evaluated by Western blot analysis.
Results:
The COX-2 siRNA fragment could be successfully transfected into RPMI8226 cells, which resulted in the significant inhibition of transcription and expression of COX-2 in the myeloma cells. Proliferation of the transfected cells was inhibited and apoptosis was induced (6.52%±0.32%, 12.53%±2.52%, 24.39%±3.51% and 36.48%±4.96% for 0, 12, 24, and 48 h, respectively) in a time-dependent manner (P<0.01). However, the expression of Bcl-2 and Bax in the RPMI8226 cells had no significant changes after nucleofection.
Conclusion:
COX-2 siRNA transfection can suppress COX-2 expression in human myeloma RPMI8226 cells, which leads to growth inhibition and apoptosis independent of Bcl-2.
Similar content being viewed by others
Article PDF
References
Hellek M, Bergsagel PL, Anderson KC . Multiple myeloma: increasing evidence for a multistep transformation process. Blood 1998; 91: 3–21.
Hideshima T, Chauhan D, Ishitsuka K, Yasui H, Raje N, Kumar S, et al. Molecular characterization of PS-341 (bortezomib) resistance: implications for overcoming resistance using lysophosphatidic acid acyltransferase (LPAAT)-β inhibitors. Oncogene 2005; 24: 3121–9.
Mitsiades N, Mitsiades C, Poulaki V, Chauhan D, Richardson P, Hideshima T, et al. Biologic sequelae of nuclear factor-kB blockade in multiple myeloma: therapeutic applications. Blood 2002; 99: 4079–86.
Pikarsky E, Porat RM, Stein I, Abramovitch R, Amit S, Kasem S, et al. NF-KappaB functions as a tumour promoter in inflammation-associated cancer. Nature 2004; 431: 461–6.
Zhang DY, Wu J, Ye F, Xue L, Jiang S, Yi J, et al. Inhibition of cancer cell proliferation and prostaglandin E2 synthesis by Scutellaria baicalensis. Cancer Res 2003; 63: 4037–43.
Anderson GD, Hauser SD, McGarity KL, Bremer ME, Isakson PC, Gregory SA . Selective inhibition of cyclooxygenase (COX)-2 reverses inflammation and expression of COX-2 and interleukin 6 in rat adjuvant arthritis. J Clin Invest 1996; 97: 2672–9.
Morita I . Distinct function of COX-1 and COX-2. Prostaglandins Other Lipid Mediat 2002; 68–69: 165–75.
Pai R, Soreghan B, Szabo IL, Pavelka M, Baatar D, Tarnawski AS . Prostaglandin E2 transactivates EGF receptor: a novel mechanism for promoting colon cancer growth and gastrointestinal hypertrophy. Nat Med 2002; 8: 289–93.
Chan G, Boyle JO, Yang EK, Zhang F, Sacks PG, Shah JP, et al. Cyclooxygenase-2 expression is up-regulated in squamous cell carcinoma of the head and neck. Cancer Res 1999; 59: 991–4.
Kawano M, Hirano T, Matsuda T, Taga T, Horii Y, Iwato K, et al. Autocrine generation and requirement of BSF-2/IL-6 for human multiple myelomas. Nature 1988; 332: 83–5.
Ladetto M, Vallet S, Trojan A, Maria D, Monitillo L, Rosato R, et al. Cyclooxygenase-2 (COX-2) is frequently expressed in multiple myeloma and is an independent predictor of poor outcome. Blood 2005; 105: 4784–91.
Trojan A, Tinguely M, Vallet S, Seifert B, Jenni B, Zippelius A, et al. Clinical significance of cyclooxygenase-2 (COX-2) in multiple myeloma. Swiss Med Wkly 2006; 136: 400–3.
Aluigi M, Fogli M, Curti A, Isidori A, Gruppioni E, Chiodoni C, et al. Nucleofection is an efficient nonviral transfection technique for human bone marrow-derived mesenchymal stem cells. Stem Cells 2006; 24: 454–61.
Zhi HY, Wang L, Zhang J, Zhou C, Ding F, Luo A, et al. Significance of COX-2 expression in human esophageal squamous cell carcinoma. Carcinogenesis 2006; 27: 1214–21.
Yan Y, Li JX, Ouyang WM, Ma Q, Hu Y, Zhang DY, et al. NFAT3 is specifically required for TNF-α-induced cyclooxygenase-2 (COX-2) expression and transformation of C141 cells. J Cell Sci 2006; 119: 2985–94.
Shin YK, Park JS, Kim HS, Jun HJ, Kim GE, Suh CO, et al. Radiosensitivity enhancement by celecoxib, a cyclooxygenase (COX)-2 selective inhibitor, via COX-2-dependent cell cycle regulation on human cancer cells expressing differential COX-2 levels. Cancer Res 2005; 65: 9501–9.
Tsujii M, DuBois R . Alterations in cellular adhesion and apptosis in epithelial cells overexpressing prostaglandin endoperoxide syntherase 2. Cell 1995; 83: 493–501.
Wu J, Xia XH, Tu PS, Fan DM, Lin CM, Kung HF, et al. 15-Lipoxygenase-1 mediates cyclooxygenase-2 inhibitor-induced apoptosis in gastric cancer. Carcinogenesis 2003; 24: 243–47.
Li QB, You Y, Chen ZC, Lv J, Shao J, You Y, et al. Role of Baicalein in the regulation of proliferation and apoptosis in human myeloma RPMI8226 cells. Chin Med J 2006; 119: 948–52.
Prince HM, Mileshkin L, Roberts A, Ganju V, Underhill C, Catalano J, et al. A Multicenter Phase II Trial of Thalidomide and Celecoxib for Patients with Relapsed and Refractory Multiple Myeloma. Clin Cancer Res 2005; 11: 5504–14.
Lacroix S, Rivest S . Effect of acute systemic inflammatory response and cytokines on the transcription of the genes encoding cyclooxygenase enzyme (COX-1 and COX-2) in the rat brain. J Neurochem 1998; 70: 452–66.
Costes V, Portier M, Lu ZY, Rossi JF, Bataille R, Klein B . Interleukin-1 in multiple myeloma: producer cells and their role in the control of IL-6 production. Br J Haematol 1998; 103: 1152–60.
Zhang M, Abe Y, Matsushima T, Nishimura J, Nawata H, Muta K . Selective cyclooxygenase-2 inhibitor NS-398 induces apoptosis in myeloma cells via a Bcl-2 independent pathway. Leuk Lymphoma 2005; 46: 425–33.
Kawane K, Fukuyama H, Yoshida H, Nagase H, Ohsawa Y, Uchiyama Y, et al. Impaired thymic development in mouse embryos deficient in apoptotic DNA degradation. Nat Immunol 2003; 4: 138–44.
Panaretakis T, Pokrovskaja K, Shoshan MC, Grander D . Interferon-alpha-induced apoptosis in U266 cells is associated with activation of the proapoptotic Bcl-2 family members Bak and Bax. Oncogene 2003; 22: 4543–56.
Oltvai ZN, Milliman CL, Korsmeyer K . Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 1993; 74: 609–19.
Sheng H, Shao J, Morrow JD, Beauchamp RD, Dubois RN . Modulation of apoptosis and Bcl-2 expression by prostaglandin E2 in human colon cancer cells. Cancer Res 1998; 58: 362–6.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Li, Qb., Chen, Zc., You, Y. et al. Small interfering RNA of cyclooxygenase-2 induces growth inhibition and apoptosis independently of Bcl-2 in human myeloma RPMI8226 cells. Acta Pharmacol Sin 28, 1031–1036 (2007). https://doi.org/10.1111/j.1745-7254.2007.00550.x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1111/j.1745-7254.2007.00550.x
Keywords
This article is cited by
-
COX2 confers bone marrow stromal cells to promoting TNFα/TNFR1β-mediated myeloma cell growth and adhesion
Cellular Oncology (2021)
-
Characterization of microRNAs regulating cyclooxygenase-2 gene expression
Genes & Genomics (2011)
-
shRNA-targeted Cyclooxygenase (COX)-2 inhibits proliferation, reduces invasion and enhances chemosensitivity in laryngeal carcinoma cells
Molecular and Cellular Biochemistry (2008)