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Article

Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line

  • Authors:
    • Yuan Zhou
    • Han-Dong Wang
    • Lin Zhu
    • Zi-Xiang Cong
    • Ning Li
    • Xiang-Jun Ji
    • Hao Pan
    • Jia-Wei Wang
    • Wan-Chun Li
  • View Affiliations / Copyright

    Affiliations: Department of Neurosurgery, Jinling Hospital, School of Medicine, Nanjing University, Nanjing 210002, P.R. China, Department of Neurosurgery, School of Medicine, Second Military Medical University (Shanghai), Jinling Hospital, Jiangsu, Nanjing 210002, P.R. China, Department of Pathology, Jinling Hospital, School of Medicine, Nanjing University, Nanjing 210002, P.R. China
  • Pages: 394-400
    |
    Published online on: October 31, 2012
       https://doi.org/10.3892/or.2012.2115
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Abstract

Glioblastoma multiforme (GBM) and oxidative stress are closely linked. Oxidative stress affects many signaling pathways and may cause the induction of autophagy. The NF-E2-related factor 2 (Nrf2)/Kelch-like ECH-associated protein 1 (Keap1) signaling pathway is the main pathway responsible for cell defense against oxidative stress and Nrf2 is a critical transcription factor related with cancer multidrug resistance. However, the relation between Nrf2 and regulation of autophagy is not well understood. In this study, we used temozolomide (TMZ), which inhibited the viability of GBM cells mainly by inducing autophagic cell death and explored the role of Nrf2 downregulation on autophagy induced by TMZ in GBM cells. In U251-Si-Nrf2 48 h after transfection the protein levels of Nrf2 were significantly downregulated, while the protein levels of LC3B-II increased by western blot analysis. Knockdown of Nrf2 also led to a significant increase of autophagic vacuoles and acidic vesicular organelles (AVOs), revealed by trans­mission electron microscopy (TEM) and acridine orange (AO) staining using flow cytometry. Collectively, these findings demonstrate that knockdown of Nrf2 can enhance the basal level of autophagy in the U251 glioma cell line. Furthermore, after the treatment with TMZ (100 µM) for 3 days, the U251-Si-Nrf2 transfected cells showed less viability rate by cell counting kit-8 (CCK-8) assay and the levels of autophagy increased obviously through analysis of western blot and AO staining using flow cytometry. Taken together, our results suggest that knockdown of Nrf2 may enhance autophagy induced by TMZ in the U251 glioma cell line, which should be further evaluated for novel anticancer activity.
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1 

Germano IM, Emdad L, Qadeer ZA, Binello E and Uzzaman M: Embryonic stem cell (ESC)-mediated transgene delivery induces growth suppression, apoptosis and radiosensitization, and overcomes temozolomide resistance in malignant gliomas. Cancer Gene Ther. 17:664–674. 2010. View Article : Google Scholar

2 

Ulasov IV, Sonabend AM, Nandi S, Khramtsov A, Han Y and Lesniak MS: Combination of adenoviral virotherapy and temozolomide chemotherapy eradicates malignant glioma through autophagic and apoptotic cell death in vivo. Br J Cancer. 100:1154–1164. 2009. View Article : Google Scholar : PubMed/NCBI

3 

Stupp R, Mason WP, van den Bent MJ, et al: Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. New Engl J Med. 352:987–996. 2005. View Article : Google Scholar : PubMed/NCBI

4 

Jiang H, White EJ, Conrad C, Gomez-Manzano C and Fueyo J: Autophagy pathways in glioblastoma. Method Enzymol. 453:273–286. 2009. View Article : Google Scholar : PubMed/NCBI

5 

Pan H, Wang H, Zhu L, Mao L, Qiao L and Su X: The Role of Nrf2 in migration and invasion of human glioma cell U251. World Neurosurg. Nov 7–2011.(Epub ahead of print).

6 

Zhang DD: The Nrf2-Keap1-ARE signaling pathway: The regulation and dual function of Nrf2 in cancer. Antioxid Redox Signal. 13:1623–1626. 2010. View Article : Google Scholar : PubMed/NCBI

7 

Lau A, Villeneuve NF, Sun Z, Wong PK and Zhang DD: Dual roles of Nrf2 in cancer. Pharmacol Res. 58:262–270. 2008. View Article : Google Scholar : PubMed/NCBI

8 

Akhdar H, Loyer P, Rauch C, Corlu A, Guillouzo A and Morel F: Involvement of Nrf2 activation in resistance to 5-fluorouracil in human colon cancer HT-29 cells. Eur J Cancer. 45:2219–2227. 2009. View Article : Google Scholar : PubMed/NCBI

9 

Zhang J, Stevens MF and Bradshaw TD: Temozolomide: mechanisms of action, repair and resistance. Curr Mol Pharmacol. 5:102–114. 2012. View Article : Google Scholar : PubMed/NCBI

10 

Wang XJ, Sun Z, Villeneuve NF, et al: Nrf2 enhances resistance of cancer cells to chemotherapeutic drugs, the dark side of Nrf2. Carcinogenesis. 29:1235–1243. 2008. View Article : Google Scholar : PubMed/NCBI

11 

Levine B, Mizushima N and Virgin HW: Autophagy in immunity and inflammation. Nature. 469:323–335. 2011. View Article : Google Scholar : PubMed/NCBI

12 

Klionsky DJ, Abeliovich H, Agostinis P, et al: Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy. 4:151–175. 2008. View Article : Google Scholar

13 

Mathew R and White E: Autophagy in tumorigenesis and energy metabolism: friend by day, foe by night. Curr Opin Genet Dev. 21:113–119. 2011. View Article : Google Scholar : PubMed/NCBI

14 

Jeon SH, Kim SH, Kim Y, et al: The tricyclic antidepressant imipramine induces autophagic cell death in U-87MG glioma cells. Biochem Biophys Res Commun. 413:311–317. 2011. View Article : Google Scholar : PubMed/NCBI

15 

Park EJ, Choi KS and Kwon TK: β-Lapachone-induced reactive oxygen species (ROS) generation mediates autophagic cell death in glioma U87 MG cells. Chem Biol Interact. 189:37–44. 2011.

16 

Jiang H, Gomez-Manzano C, Aoki H, et al: Examination of the therapeutic potential of Delta-24-RGD in brain tumor stem cells: role of autophagic cell death. J Natl Cancer Inst. 99:1410–1414. 2007. View Article : Google Scholar : PubMed/NCBI

17 

Kaza N, Kohli L and Roth KA: Autophagy in brain tumors: a new target for therapeutic intervention. Brain Pathol. 22:89–98. 2012. View Article : Google Scholar : PubMed/NCBI

18 

Jakubowicz-Gil J, Langner E and Rzeski W: Kinetic studies of the effects of Temodal and quercetin on astrocytoma cells. Pharmacol Rep. 63:403–416. 2011. View Article : Google Scholar : PubMed/NCBI

19 

Kanzawa T, Germano IM, Komata T, Ito H, Kondo Y and Kondo S: Role of autophagy in temozolomide-induced cytotoxicity for malignant glioma cells. Cell Death Differ. 11:448–457. 2004. View Article : Google Scholar : PubMed/NCBI

20 

Natsumeda M, Aoki H, Miyahara H, et al: Induction of autophagy in temozolomide treated malignant gliomas. Neuropathology. 31:486–493. 2011. View Article : Google Scholar : PubMed/NCBI

21 

Katayama M, Kawaguchi T, Berger MS and Pieper RO: DNA damaging agent-induced autophagy produces a cytoprotective adenosine triphosphate surge in malignant glioma cells. Cell Death Differ. 14:548–558. 2007. View Article : Google Scholar

22 

Kato T, Natsume A, Toda H, et al: Efficient delivery of liposome-mediated MGMT-siRNA reinforces the cytotoxity of temozolomide in GBM-initiating cells. Gene Ther. 17:1363–1371. 2010. View Article : Google Scholar : PubMed/NCBI

23 

Voss V, Senft C, Lang V, et al: The pan-Bcl-2 inhibitor (−)-gossypol triggers autophagic cell death in malignant glioma. Mol Cancer Res. 8:1002–1016. 2010.

24 

Lefranc F: Glioblastomas are resistant to apoptosis but less resistant to the autophagic process. Bull Mem Acad R Med Belg. 162:331–338. 2007.(In French).

25 

Stepkowski TM and Kruszewski MK: Molecular cross-talk between the NRF2/KEAP1 signaling pathway, autophagy, and apoptosis. Free Radic Biol Med. 50:1186–1195. 2011. View Article : Google Scholar : PubMed/NCBI

26 

Gao S, Yang XJ, Zhang WG, Ji YW and Pan Q: Mechanism of thalidomide to enhance cytotoxicity of temozolomide in U251-MG glioma cells in vitro. Chin Med J. 122:1260–1266. 2009.PubMed/NCBI

27 

Klingelhoeffer C, Kammerer U, Koospal M, et al: Natural resistance to ascorbic acid induced oxidative stress is mainly mediated by catalase activity in human cancer cells and catalase-silencing sensitizes to oxidative stress. BMC Complement Altern Med. 12:612012. View Article : Google Scholar

28 

Lau AT, Wang Y and Chiu JF: Reactive oxygen species: current knowledge and applications in cancer research and therapeutic. J Cell Biochem. 104:657–667. 2008. View Article : Google Scholar : PubMed/NCBI

29 

Bjorkoy G, Lamark T, Brech A, et al: p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol. 171:603–614. 2005. View Article : Google Scholar : PubMed/NCBI

30 

Mizushima N and Yoshimori T: How to interpret LC3 immunoblotting. Autophagy. 3:542–545. 2007. View Article : Google Scholar : PubMed/NCBI

31 

Aoki H, Kondo Y, Aldape K, et al: Monitoring autophagy in glioblastoma with antibody against isoform B of human microtubule-associated protein 1 light chain 3. Autophagy. 4:467–475. 2008. View Article : Google Scholar : PubMed/NCBI

32 

Lin CJ, Lee CC, Shih YL, et al: Resveratrol enhances the therapeutic effect of temozolomide against malignant glioma in vitro and in vivo by inhibiting autophagy. Free Radic Biol Med. 52:377–391. 2012. View Article : Google Scholar : PubMed/NCBI

33 

Chen H-Y and White E: Role of autophagy in cancer prevention. Cancer Prev Res. 4:973–983. 2011. View Article : Google Scholar : PubMed/NCBI

34 

Moreau K, Luo S and Rubinsztein DC: Cytoprotective roles for autophagy. Curr Opin Cell Biol. 22:206–211. 2010. View Article : Google Scholar : PubMed/NCBI

35 

Rao VA, Klein SR, Bonar SJ, et al: The antioxidant transcription factor Nrf2 negatively regulates autophagy and growth arrest induced by the anticancer redox agent mitoquinone. J Biol Chem. 285:34447–34459. 2010. View Article : Google Scholar : PubMed/NCBI

36 

Pan Q, Yang XJ, Wang HM, et al: Chemoresistance to temozolomide in human glioma cell line U251 is associated with increased activity of O6-methylguanine-DNA methyltransferase and can be overcome by metronomic temozolomide regimen. Cell Biochem Biophys. 62:185–191. 2012. View Article : Google Scholar : PubMed/NCBI

37 

Cho JM, Manandhar S, Lee HR, Park HM and Kwak MK: Role of the Nrf2-antioxidant system in cytotoxicity mediated by anticancer cisplatin: implication to cancer cell resistance. Cancer Lett. 260:96–108. 2008. View Article : Google Scholar : PubMed/NCBI

38 

Nezis IP and Stenmark H: p62 at the interface of autophagy, oxidative stress signaling, and cancer. Antioxid Redox Signal. 17:786–793. 2012. View Article : Google Scholar : PubMed/NCBI

39 

Inami Y, Waguri S, Sakamoto A, et al: Persistent activation of Nrf2 through p62 in hepatocellular carcinoma cells. J Cell Biol. 193:275–284. 2011. View Article : Google Scholar : PubMed/NCBI

40 

Jain A, Lamark T, Sjottem E, et al: p62/SQSTM1 is a target gene for transcription factor NRF2 and creates a positive feedback loop by inducing antioxidant response element-driven gene transcription. J Biol Chem. 285:22576–22591. 2010. View Article : Google Scholar : PubMed/NCBI

41 

Komatsu M, Kurokawa H, Waguri S, et al: The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1. Nat Cell Biol. 12:213–223. 2010.PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Zhou Y, Wang H, Zhu L, Cong Z, Li N, Ji X, Pan H, Wang J and Li W: Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line. Oncol Rep 29: 394-400, 2013.
APA
Zhou, Y., Wang, H., Zhu, L., Cong, Z., Li, N., Ji, X. ... Li, W. (2013). Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line. Oncology Reports, 29, 394-400. https://doi.org/10.3892/or.2012.2115
MLA
Zhou, Y., Wang, H., Zhu, L., Cong, Z., Li, N., Ji, X., Pan, H., Wang, J., Li, W."Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line". Oncology Reports 29.1 (2013): 394-400.
Chicago
Zhou, Y., Wang, H., Zhu, L., Cong, Z., Li, N., Ji, X., Pan, H., Wang, J., Li, W."Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line". Oncology Reports 29, no. 1 (2013): 394-400. https://doi.org/10.3892/or.2012.2115
Copy and paste a formatted citation
x
Spandidos Publications style
Zhou Y, Wang H, Zhu L, Cong Z, Li N, Ji X, Pan H, Wang J and Li W: Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line. Oncol Rep 29: 394-400, 2013.
APA
Zhou, Y., Wang, H., Zhu, L., Cong, Z., Li, N., Ji, X. ... Li, W. (2013). Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line. Oncology Reports, 29, 394-400. https://doi.org/10.3892/or.2012.2115
MLA
Zhou, Y., Wang, H., Zhu, L., Cong, Z., Li, N., Ji, X., Pan, H., Wang, J., Li, W."Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line". Oncology Reports 29.1 (2013): 394-400.
Chicago
Zhou, Y., Wang, H., Zhu, L., Cong, Z., Li, N., Ji, X., Pan, H., Wang, J., Li, W."Knockdown of Nrf2 enhances autophagy induced by temozolomide in U251 human glioma cell line". Oncology Reports 29, no. 1 (2013): 394-400. https://doi.org/10.3892/or.2012.2115
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