Estrogen Receptor Alpha 36 Gene Knockdown Promote the Expression of NF-κB in PC12 Cells


The nuclear transcription factors κB (NF-κB) is widely existing in various kinds of cell types in the nervous system and plays an important role in neuron apoptosis and neurodegenerative diseases. Estrogen receptor alpha 36 (ER-α36), is a novel variant of ERα (as known ER-α66) which can transduce both estrogenand antiestrogen-dependent activation of MAPK signal pathway and stimulate cell growth. Here, we aimed to detect the effect of ER-α36 gene silencing on the expression of NF-κB in normal cultured PC12 cells and to provide an experimental foundation for understanding the function of ER-α36 innerve cells. PC12 cells with ER-α36 expression knocked down by the shRNA method. Then Western blot and immunocytochemical staining were performed to detect the expression and translocation of NF-κB after transfection. The results showed that NF-κB expression was significantly higher comparing with the control group after transfection (P < 0.01). Also, NF-κB subunit entered nuclear after transfection; Immunofluorescence staining and immunocytochemical staining of PC12 cells demonstrated that ER-α36 was expressed mainly on the plasma membrane and on the cell nucleus membrane. These data indicate that ER-α36 gene silencing can increase the expression of NF-κB and promote its nuclear translocation in PC12 cells.

Share and Cite:

P. Zou, C. Qu, Y. Xu, H. Li, D. Han, D. Shi and W. Zou, "Estrogen Receptor Alpha 36 Gene Knockdown Promote the Expression of NF-κB in PC12 Cells," Open Journal of Endocrine and Metabolic Diseases, Vol. 3 No. 4B, 2013, pp. 20-24. doi: 10.4236/ojemd.2013.34A2003.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] F. Cavttaneo, G. Guerra and R. Ammendola, “Expression and Signaling of Formyl-Peptide Receptors in the Brain,” Neurochemical Research, Vol. 35, No. 12, 2010, pp. 2018-2026. doi:10.1007/s11064-010-0301-5
[2] I. Charalampopoulos, V. I. Alexaki, C. Tsatsanis, V. Minas, E. Dermitzaki, I. Lasaridis, L. Vardouli, C. Stournaras, A. N. Margioris, E. Castanas and A. Gravanis, “Neurosteroids as Endogenous Inhibitors of Neuronal Cell Apoptosis in Aging,” Annals of the New York Academy of Sciences, Vol. 1088, 2006, pp. 139-152. doi:10.1196/annals.1366.003
[3] C. H. Nijboer, C. J. Heijnen, F. Groenendaal, M. J. May, F. van Bel and A. Kavelaars, “Strong Neuroprotection by Inhibition of NF-KappaB after Neonatal Hypoxia-Ischemia Involves Apoptotic Mechanisms but Is Independent of Cytokines,” Stroke, Vol. 39, No. 7, 2008, pp. 2129-2137. doi:10.1161/STROKEAHA.107.504175
[4] S. M. Fernandez, M. C. Lewis, A. S. Pechenino, L. L. Harburger, P. T. Orr, J. E. Gresack, G. E. Schafe and K. M. Frick, “Estradiol-Induced Enhancement of Object Memory Consolidation Involves Hippocampal Extracellular Signal-Regulated Kinase Activation and MembraneBound Estrogen Receptors,” The Journal of Neuroscience, Vol. 28, No. 35, 2008, pp. 8660-8667. doi:10.1523/JNEUROSCI.1968-08.2008
[5] Z. Wang, X. Zhang, P. Shen, B. W. Loggie, Y. Chang and T. F. Deuel, “Identification, Cloning, and Expression of Human Estrogen Receptor-Alpha36, a Novel Variant of Human Estrogen Receptor-Alpha66,” Biochemical and Biophysical Research Communications, Vol. 336, No. 4, 2005, pp. 1023-1027. doi:10.1016/j.bbrc.2005.08.226
[6] L. M. Lee, J. Cao, H. Deng, P. Chen, Z. Gatalica and Z. Y. Wang, “ER-Alpha36, a Novel Variant of ER-Alpha, Is Expressed in ER-Positive and -Negative Human Breast Carcinomas,” Anticancer Research, Vol. 28, No. 1B, 2008, pp. 479-483.
[7] C. Hicks, R. Kumar, A. Pannuti and L. Miele, “Integrative Analysis of Response to Tamoxifen Treatment in ER-Positive Breast Cancer Using GWAs Information and Transcription Profiling,” Breast Cancer, Vol. 6, 2012, pp. 47-66.
[8] K. W. Nettles, G. Gil, J. Nowak, R. Métivier, V. B. Sharma and G. L. Greene, “CBP Is a Dosage-Dependent Regulator of Nuclear Factor—KappaB Suppression by the Estrogen Receptor,” Molecular Endocrinology, Vol. 22, No. 2, 2008, pp. 263-272. doi:10.1210/me.2007-0324
[9] J. W. Antoon, M. D. White, E. M. Slaughter, J. L. Driver, H. S. Khalili, S. Elliott, C. D. Smith, M. E. Burow and B.S. Beckman, “Targeting NFκB Mediated Breast Cancer Chemoresistance through Selective Inhibition of Sphingosine Kinase-2,” Cancer Biology & Therapy, Vol. 11, No. 7, 2011, pp. 678-689. doi:10.4161/cbt.11.7.14903
[10] Y. Zhou, S. Eppenberger-Castori, C. Marx, C. Yau, G. K. Scott, U. Eppenberger and C. C. Benz, “Activation of Nuclear Faetor-KappaB (NFkappaB) Identifies a High-Risk Subset of Hormone-Dependent Breast Cancers,” The International Journal of Biochemistry & Cell Biology, Vol. 37, No. 5, 2005, pp. l130-l144. doi:10.1016/j.biocel.2004.09.006
[11] L. Shi, B. Dong, Z. Li, Y. Lu, T. Ouyang, J. Li, T. Wang, Z. Fan, T. Fan, B. Lin, Z. Wang and Y. Xie, “Expression of ER-(α)36, a Novel Variant of Estrogen Receptor-(α), and Resistance to Tamoxifen Treatment in Breast Cancer,” Journal of Clinical Oncology, Vol. 27, No. 21, 2009, pp. 3423-3429. doi:10.1200/JCO.2008.17.2254
[12] G. Zhang and S. Ghosh, “Toll-Like Receptor-Mediated NFκB Activation: A Phylogenetically Conserved Paradigm in Innate Immunity,” Journal of Clinical Investigation, Vol. 107, No. 1, 2001, pp. 13-19. doi:10.1172/JCI11837
[13] A. L. Bernardino, T. A. Myers, X. Alvarez, A. Hasegawa and M. T. Philipp, “Toll-Like Receptors: Insights into Their Possible Role in the Pathogenesis of Lyme Neuroborreliosis,” Infection and Immunity, Vol. 76, No. 10, 2008, pp. 4385-4395. doi:10.1128/IAI.00394-08
[14] J. K. Lee, J. Chung, K. M. Druey and M. G. Tansey, “RGS10 Exerts a Neuroprotective Role through the PKA/ c-AMP ResponseElement (CREB) Pathway in Dopaminergic Neuron-Like Cells,” Journal of Neurochemistry, Vol. 122, No. 2, 2012, pp. 333-343.
[15] L. I. McKay and J. A. Cidlowski, “Molecular Control of Immune/Inflammatory Responses: Interactions between Nuclear Factor-Kappa B and Steroid Receptor-Signaling Pathways,” Endocrine Reviews, Vol. 20, No. 4, 1999, pp. 435-459. doi:10.1210/er.20.4.435
[16] N. Maulik, M. Sato, B. D. Price and D. K. Das, “An Essential Role of NFkappaB in Tyrosine Kinase Signaling of p38 MAP Kinase Regulation of Myocardial Adaptation to Ischemia,” FEBS Letters, Vol. 429, No. 3, 1998, pp. 365-369. doi:10.1016/S0014-5793(98)00632-2
[17] I. Sarnico, A. Lanzillotta, M. Benarese, M. Alghisi, C. Baiguera, L. Battistin, P. Spano and M. Pizzi, “NF-KappaB Dimers in the Regulation of Neuronal Survival,” International Review of Neurobiology, Vol. 85, 2009, pp. 351-362. doi:10.1016/S0074-7742(09)85024-1
[18] C. H. Nijboer, C. J Heijnen, F. Groenendaal, M. J. May, F. Bel and A. Kavelaars, “Strong Neuroprotection by Inhibition of NF-KappaB after Neonatal Hypoxia-Ischemia Involves Apoptotic Mechanisms but Is Independent of Cytokines,” Stroke, Vol. 39, No. 7, 2008, pp. 2129-2137. doi:10.1161/STROKEAHA.107.504175
[19] C. B. Weldon, A. P. Parker, D. Patten, S. Elliott, Y. Tang, D. E. Frigo, C. M. Dugan, E. L. Coakley, N. N. Butler, J. L. Clayton, J. Alam, T. J. Curiel, B. S. Beckman, B. M. Jaffe and M. E. Burow, “Sensitization of ApoptoticallyResistant Breast Carcinoma Cells to TNF and TRAIL by Inhibition of p38 Mitogen-Activated Protein Kinase Signaling,” International Journal of Oncology, Vol. 24, No. 6, 2004, pp. 1473-1480.
[20] J. Zhang, G. Li, Z. Li, X. Yu, Y. Zheng, K. Jin, H. Wang, Y. Gong, X. Sun, X. Teng, J. Cao and L. Teng, “Estrogen-Independent Effects of ER-a36 in ER-Negative Breast Cancer,” Steroids, Vol. 77, No. 6, 2012, pp. 666-673. doi:10.1016/j.steroids.2012.02.013
[21] X. T. Zhang, L. Ding, L. G. Kang and Z. Y. Wang, “Involvement of ER-α36, Src, EGFR and STAT5 in the Biphasic Estrogen Signaling of ER-Negative Breast Cancer Cells,” Oncology Reports, Vol. 27, No. 6, 2012, pp. 2057-2065.
[22] X. Zhang, L. Ding, L. Kang and Z. Y. Wang, “Estrogen Receptor-Alpha 36 Mediates Mitogenic Antiestrogen Signaling in ER-Negative Breast Cancer Cells,” PLOS One, Vol. 7, No. 1, 2012, pp. e30174-30186.
[23] L. Kang, Y. Guo, X. Zhang, J. Meng and Z. Y. Wang, “A Positive Cross-Regulation of HER2 and ER-α36 Controls ALDH1 Positive Breast Cancer Cells,” Molecular Biology, Vol. 127, No. 3-5, 2011, pp. 262-268.
[24] J. Rao, X. Jiang, Y. Wang and B. Chen, “Advances in the Understanding of the Structure and Function of ER-α36, a Novel Variant of Human Estrogen Receptor-Alpha,” Molecular Biology, Vol. 127, No. 3-5, 2011, pp. 231-237.
[25] M. E. Quaedackers, C. E. van den Brink, P. T. van der Saag and L. G. Tertoolen, “Tertoolen LG Direct Interaction between Estrogen Receptor Alpha and NF-KappaB in the Nucleus of Living Cells,” Molecular and Cellular Endocrinology, Vol. 273, No. 1-2, 2007, pp. 42-50. doi:10.1016/j.mce.2007.05.002

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.