DNA methylation-mediated caspase-8 downregulation is associated with anti-apoptotic activity and human malignant glioma grade

Teng,Yueqiu; Dong,Yu‑Cui; Liu,Zhaoyu; Zou,Yan; Xie,Hui; Zhao,Yu; Su,Jun; Cao,Fenglin; Jin,Hua; Ren,Huan

doi:10.3892/ijmm.2017.2881

DNA methylation-mediated caspase-8 downregulation is associated with anti-apoptotic activity and human malignant glioma grade

Authors:
- Yueqiu Teng
- Yu‑Cui Dong
- Zhaoyu Liu
- Yan Zou
- Hui Xie
- Yu Zhao
- Jun Su
- Fenglin Cao
- Hua Jin
- Huan Ren
View Affiliations

Affiliations: Department of Immunology, Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China, Department of Neurosurgery, The Third Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150040, P.R. China, Central Laboratory of Blood Cancer, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: February 7, 2017 https://doi.org/10.3892/ijmm.2017.2881
Pages: 725-733

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

The methylation-mediated silencing of tumor suppressors, including key apoptosis-related genes plays an important role in the pathogenesis and therapeutic resistance in human cancer. In this study, we aimed to elucidate the role and mechanisms of resistance to apoptosis with caspase-8 gene downregulation in human malignant glioma. Reverse transcription-polymerase chain reaction (RT-PCR) and methylation-specific PCR (MSP) were used to examine caspase-8 expressoin at the mRNA level and gene methylation status in normal brain tissue, glioma tissue and cancer cell lines. Caspase-8 protein kinase activity was measured by caspase-8 colorimetric assays; cell apoptosis was examined by Annexin V/propidium iodide (PI) staining; the rates of tumor cell apoptosis were detected by flow cytometry. Our results revealed that caspase-8 gene silencing may result from the methylation of its gene promoter in human glioma tissues. The expression of caspase-8 at the mRNA level was significantly associated with the grade of human glioma. In certain human cancer cell lines, the expression at the mRNA level, protein kinase activity and tumor cell anti-apoptotic activity and resistance were related to the methylation status of the caspase-8 gene promoter. Thus, the caspase-8 gene methylation status may be used as an indicator for the early diagnosis of human malignant glioma. Combination therapy with demethylation reagents may overcome therapeutic resistance in the same malignancy.

View Figures

View References

1	Fuller GN and Scheithauer BW: Symposium: The 2007 revised world health organization (WHO) Classification of tumors of the central nervous system: newly codified entities. Brain Pathol. 17:304–307. 2007. View Article : Google Scholar : PubMed/NCBI
2	Thompson CB: Apoptosis in the pathogenesis and treatment of disease. Science. 267:1456–1462. 1995. View Article : Google Scholar : PubMed/NCBI
3	Varela M, Ranuncolo SM, Morand A, Lastiri J, De Kier Joffé EB, Puricelli LI and Pallotta MG: EGF-R and PDGF-R, but not bcl-2, overexpression predict overall survival in patients with low-grade astrocytomas. J Surg Oncol. 86:34–40. 2004. View Article : Google Scholar : PubMed/NCBI
4	Kajiwara Y, Yamasaki F, Hama S, Yahara K, Yoshioka H, Sugiyama K, Arita K and Kurisu K: Expression of survivin in astrocytic tumors: Correlation with malignant grade and prognosis. Cancer. 97:1077–1083. 2003. View Article : Google Scholar : PubMed/NCBI
5	Sarkar C, Karak AK, Nath N, Sharma MC, Mahapatra AK, Chattopadhyay P and Sinha S: Apoptosis and proliferation: Correlation with p53 in astrocytic tumours. J Neurooncol. 73:93–100. 2005. View Article : Google Scholar : PubMed/NCBI
6	Grenet J, Teitz T, Wei T, Valentine V and Kidd VJ: Structure and chromosome localization of the human CASP8 gene. Gene. 226:225–232. 1999. View Article : Google Scholar : PubMed/NCBI
7	Boldin MP, Goncharov TM, Goltsev YV and Wallach D: Involvement of MACH, a novel MORT1/FADD-interacting protease, in Fas/APO-1- and TNF receptor-induced cell death. Cell. 85:803–815. 1996. View Article : Google Scholar : PubMed/NCBI
8	Muzio M, Chinnaiyan AM, Kischkel FC, O'Rourke K, Shevchenko A, Ni J, Scaffidi C, Bretz JD, Zhang M, Gentz R, et al: FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death--inducing signaling complex. Cell. 85:817–827. 1996. View Article : Google Scholar : PubMed/NCBI
9	Deng Y, Lin Y and Wu X: TRAIL-induced apoptosis requires Bax-dependent mitochondrial release of Smac/DIABLO. Genes Dev. 16:33–45. 2002. View Article : Google Scholar : PubMed/NCBI
10	Blanchard H, Donepudi M, Tschopp M, Kodandapani L, Wu JC and Grütter MG: Caspase-8 specificity probed at subsite S(4): Crystal structure of the caspase-8-Z-DEVD-cho complex. J Mol Biol. 302:9–16. 2000. View Article : Google Scholar : PubMed/NCBI
11	Hopkins-Donaldson S, Ziegler A, Kurtz S, Bigosch C, Kandioler D, Ludwig C, Zangemeister-Wittke U and Stahel R: Silencing of death receptor and caspase-8 expression in small cell lung carcinoma cell lines and tumors by DNA methylation. Cell Death Differ. 10:356–364. 2003. View Article : Google Scholar : PubMed/NCBI
12	McKee AE and Thiele CJ: Targeting caspase 8 to reduce the formation of metastases in neuroblastoma. Expert Opin Ther Targets. 10:703–708. 2006. View Article : Google Scholar : PubMed/NCBI
13	Ashley DM, Riffkin CD, Muscat AM, Knight MJ, Kaye AH, Novak U and Hawkins CJ: Caspase 8 is absent or low in many ex vivo gliomas. Cancer. 104:1487–1496. 2005. View Article : Google Scholar : PubMed/NCBI
14	Jones PA: DNA methylation and cancer. Oncogene. 21:5358–5360. 2002. View Article : Google Scholar : PubMed/NCBI
15	Galm O, Wilop S, Reichelt J, Jost E, Gehbauer G, Herman JG and Osieka R: DNA methylation changes in multiple myeloma. Leukemia. 18:1687–1692. 2004. View Article : Google Scholar : PubMed/NCBI
16	Sun T, Gao Y, Tan W, Ma S, Shi Y, Yao J, Guo Y, Yang M, Zhang X, Zhang Q, et al: A six-nucleotide insertion-deletion polymorphism in the CASP8 promoter is associated with susceptibility to multiple cancers. Nat Genet. 39:605–613. 2007. View Article : Google Scholar : PubMed/NCBI
17	Shen L, Kondo Y, Rosner GL, Xiao L, Hernandez NS, Vilaythong J, Houlihan PS, Krouse RS, Prasad AR, Einspahr JG, et al: MGMT promoter methylation and field defect in sporadic colorectal cancer. J Natl Cancer Inst. 97:1330–1338. 2005. View Article : Google Scholar : PubMed/NCBI
18	Shen DH, Chan KY, Khoo US, Ngan HY, Xue WC, Chiu PM, Ip P and Cheung AN: Epigenetic and genetic alterations of p33^ING1b in ovarian cancer. Carcinogenesis. 26:855–863. 2005. View Article : Google Scholar : PubMed/NCBI
19	Kim JH, Choi YD, Lee JS, Lee JH, Nam JH and Choi C: Assessment of DNA methylation for the detection of cervical neoplasia in liquid-based cytology specimens. Gynecol Oncol. 116:99–104. 2010. View Article : Google Scholar
20	Hildebrandt MA, Gu J, Lin J, Ye Y, Tan W, Tamboli P, Wood CG and Wu X: Hsa-miR-9 methylation status is associated with cancer development and metastatic recurrence in patients with clear cell renal cell carcinoma. Oncogene. 29:5724–5728. 2010. View Article : Google Scholar : PubMed/NCBI
21	Park SY, Kook MC, Kim YW, Cho NY, Jung N, Kwon HJ, Kim TY and Kang GH: CpG island hypermethylator phenotype in gastric carcinoma and its clinicopathological features. Virchows Arch. 457:415–422. 2010. View Article : Google Scholar : PubMed/NCBI
22	Kitazawa S, Kitazawa R and Maeda S: Identification of methylated cytosine from archival formalin-fixed paraffin-embedded specimens. Lab Invest. 80:275–276. 2000. View Article : Google Scholar : PubMed/NCBI
23	Hopkins-Donaldson S, Bodmer JL, Bourloud KB, Brognara CB, Tschopp J and Gross N: Loss of caspase-8 expression in highly malignant human neuroblastoma cells correlates with resistance to tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis. Cancer Res. 60:4315–4319. 2000.PubMed/NCBI
24	Hopkins-Donaldson S, Bodmer JL, Bourloud KB, Brognara CB, Tschopp J and Gross N: Loss of caspase-8 expression in neuroblastoma is related to malignancy and resistance to TRAIL-induced apoptosis. Med Pediatr Oncol. 35:608–611. 2000. View Article : Google Scholar : PubMed/NCBI
25	Teitz T, Wei T, Valentine MB, Vanin EF, Grenet J, Valentine VA, Behm FG, Look AT, Lahti JM and Kidd VJ: Caspase 8 is deleted or silenced preferentially in childhood neuroblastomas with amplification of MYCN. Nat Med. 6:529–535. 2000. View Article : Google Scholar : PubMed/NCBI
26	Saggioro FP, Neder L, Stávale JN, Paixão-Becker AN, Malheiros SM, Soares FA, Pittella JE, Matias CC, Colli BO, Carlotti CG Jr, et al: Fas, FasL, and cleaved caspases 8 and 3 in glioblastomas: A tissue microarray-based study. Pathol Res Pract. 210:267–273. 2014. View Article : Google Scholar : PubMed/NCBI
27	Jones PA and Baylin SB: The fundamental role of epigenetic events in cancer. Nat Rev Genet. 3:415–428. 2002.PubMed/NCBI
28	Gonzalez-Gomez P, Bello MJ, Inda MM, Alonso ME, Arjona D, Amiñoso C, Lopez-Marin I, de Campos JM, Sarasa JL, Castresana JS, et al: Deletion and aberrant CpG island methylation of caspase-8 gene in medulloblastoma. Oncol Rep. 12:663–666. 2004.PubMed/NCBI
29	Knudson AG Jr: Mutation and cancer: Statistical study of retinoblastoma. Proc Natl Acad Sci USA. 68:820–823. 1971. View Article : Google Scholar : PubMed/NCBI
30	Ebinger M, Senf L, Wachowski O and Scheurlen W: Promoter methylation pattern of caspase-8, P16^INK4A, MGMT, TIMP-3, and E-cadherin in medulloblastoma. Pathol Oncol Res. 10:17–21. 2004. View Article : Google Scholar
31	Adams DJ, Waud WR, Wani MC, Manikumar G, Flowers JL, Driscoll TA and Morgan LR: BACPTDP: A water-soluble camptothecin pro-drug with enhanced activity in hypoxic/acidic tumors. Cancer Chemother Pharmacol. 67:855–865. 2011. View Article : Google Scholar
32	Xia S, Lal B, Tung B, Wang S, Goodwin CR and Laterra J: Tumor microenvironment tenascin-C promotes glioblastoma invasion and negatively regulates tumor proliferation. Neuro Oncol. 18:507–517. 2016. View Article : Google Scholar