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Journal of Experimental & Clinical Cancer Research
Published in: Journal of Experimental & Clinical Cancer Research 1/2020

Open Access 01-12-2020 | Colorectal Cancer | Research

Downregulation of GLYR1 contributes to microsatellite instability colorectal cancer by targeting p21 via the p38MAPK and PI3K/AKT pathways

Authors: Zhiyan Hu, Ting Long, Yidan Ma, Jiaxian Zhu, Lingfang Gao, Yan Zhong, Xia Wang, Xiaoyan Wang, Zuguo Li

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2020

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Abstract

Background

GLYR1 has a high mutation frequency in microsatellite instability colorectal cancer (MSI CRC) and is presumed to be a novel tumor suppressor. However, the role of GLYR1 in tumors has never been studied. In particular, the downregulation of GLYR1 in MSI CRC is worthy of further investigation.

Methods

Western blot and immunohistochemistry analyses were used to detect GLYR1 protein expression in CRC tissues and cell lines, and the clinical significance of GLYR1 was also analyzed. The relationship between GLYR1 and MLH1 was validated by immunofluorescence, immunoprecipitation and bioinformatics analyses. Western blotting, qRT-PCR, CCK-8 assays, colony formation assays, flow cytometry and Hoechst 33258 staining assays were used to assess the effect of GLYR1 on the cell cycle progression, proliferation, differentiation and apoptosis of CRC cells in vitro. The related mechanisms were initially investigated by Western blotting.

Results

GLYR1 was significantly downregulated in MSI CRC and its expression was negatively correlated with tumor size and positively correlated with tumor differentiation in CRC patients. In addition, GLYR1 interacted with MLH1 to regulate its nuclear import and expression. Moreover, downregulation of GLYR1 accelerated G1/S phase transition, promoted proliferation and inhibited differentiation of SW480 and SW620 cells in vitro. Furthermore, downregulation of GLYR1 decreased the sensitivity to 5-fluorouracil (5-FU) by inhibiting the mitochondrial apoptosis pathway in CRC cells. Inhibition of the p38 mitogen-activated protein kinase (p38MAPK) and activation of the phosphatidyl 3-kinase/protein kinase B (PI3K/Akt) signaling pathways were involved in the mechanism by which GLYR1 downregulated p21.

Conclusions

Ours is the first study to elucidate the role of GLYR1 in tumors and provide evidence for GLYR1 as a biological marker that reflects the degree of malignancy and sensitivity to 5-FU in MSI CRC.
Appendix
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Literature
1.
Romanowicz-Makowska H, Smolarz B, Langner E, Kozlowska E, Kulig A, Dziki A. Analysis of microsatellite instability and BRCA1 mutations in patients from hereditary nonpolyposis colorectal cancer (HNPCC) family. Pol J PatholJournal Article; Research Support, Non-U.S. Gov’t. 2005;56(1):21–6.PubMed
2.
Iacopetta B, Grieu F, Amanuel B. Microsatellite instability in colorectal cancer. Asia Pac J Clin Oncol. Journal Article; Research Support, Non-U.S. Gov’t; Review. 2010;6(4):260–9.PubMed
3.
Strambu V, Garofil D, Pop F, Radu P, Bratucu M, Iorga C, et al. Microsatellite instability in the management of stage II colorectal patients. Chirurgia (Bucur). Journal Article; Research Support, Non-U.S. Gov’t. 2013;108(6):816–21.
4.
Jover R, Zapater P, Castells A, Llor X, Andreu M, Cubiella J, et al. The efficacy of adjuvant chemotherapy with 5-fluorouracil in colorectal cancer depends on the mismatch repair status. Eur J CancerJournal Article; Multicenter Study; Research Support, Non-U.S. Gov't. 2009;45(3):365–73.PubMed
5.
Alhopuro P, Sammalkorpi H, Niittymäki I, Biström M, Raitila A, Saharinen J, et al. Candidate driver genes in microsatellite-unstable colorectal cancer. Int J Cancer. 2012;130(7):1558–66.PubMed
6.
Shin J, Min S. Novel functional roles of caspase-related genes in the regulation of apoptosis and autophagy. Korean J Physiol Pharmacol. 2016;20(6):573.PubMedPubMedCentral
7.
Dotto GP. p21(WAF1/Cip1): more than a break to the cell cycle. Biochim Biophys ActaJournal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov’t, P.H.S.; Review. 2000;1471(1):M43.PubMed
8.
Roninson IB. Oncogenic functions of tumour suppressor p21(Waf1/Cip1/Sdi1): association with cell senescence and tumour-promoting activities of stromal fibroblasts. Cancer LettJournal Article; Research Support, U.S. Gov’t, P.H.S.; Review. 2002;179(1):1–14.PubMed
9.
Dalton S. Linking the cell cycle to cell fate decisions. Trends Cell BiolJournal Article; Research Support, N.I.H., Extramural; Review. 2015;25(10):592–600.PubMedPubMedCentral
10.
Blagosklonny MV, Pardee AB. The restriction point of the cell cycle. Cell CycleJournal Article; Review. 2002;1(2):103–10.PubMed
11.
Harper JW, Adami GR, Wei N, Keyomarsi K, Elledge SJ. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. CellJournal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov’t, P.H.S. 1993;75(4):805–16.PubMed
12.
Zager RA, Johnson AC. Acute kidney injury induces dramatic p21 up-regulation via a novel, glucocorticoid- activated, p53 independent, pathway. Am J Physiol-Renal. 2019;316(4):F674.
13.
Chen H, Shan J, Chen D, Wang R, Qi W, Wang H, et al. CtIP promotes G2/M arrest in etoposide-treated HCT116 cells in a p53-independent manner. J Cell Physiol. 2018;234(7):11871.PubMed
14.
Wang Y, Qiu C, Lu N, Liu Z, Jin C, Sun C, et al. FOXD1 is targeted by miR-30a-5p and miR-200a-5p and suppresses the proliferation of human ovarian carcinoma cells by promoting p21 expression in a p53-independent manner. Int J Oncol. 2018;52(6):2130–42.PubMed
15.
Huang S, Li Y, Yuan X, Zhao M, Wang J, Li Y, et al. The UbL-UBA Ubiquilin4 protein functions as a tumor suppressor in gastric cancer by p53-dependent and p53-independent regulation of p21. Cell Death Differ. 2019;26(3):516–30.PubMed
16.
Kim GY, Mercer SE, Ewton DZ, Yan Z, Jin K, Friedman E. The stress-activated protein kinases p38 alpha and JNK1 stabilize p21(Cip1) by phosphorylation. J Biol ChemJournal Article; Research Support, U.S. Gov’t, P.H.S. 2002;277(33):29792–802.PubMed
17.
Wei L, Lin J, Wu G, Xu W, Li H, Hong Z, et al. Scutellaria barbata D. Don induces G1/S arrest via modulation of p53 and Akt pathways in human colon carcinoma cells. Oncol RepJournal Article; Research Support, Non-U.S. Gov’t. 2013;29(4):1623–8.PubMed
18.
Guo X, Yang Y, Liu L, Liu X, Xu J, Wu K, et al. Pirfenidone induces G1 arrest in human Tenon's fibroblasts in vitro involving AKT and MAPK signaling pathways. J Ocul Pharmacol Th. 2017;33(5):366–74.
19.
Jiang Y, Gram H, Zhao M, New L, Gu J, Feng L, et al. Characterization of the structure and function of the fourth member of p38 group mitogen-activated protein kinases, p38delta. J Biol ChemJournal Article; Research Support, Non-U.S. Gov’t; Research Support, U.S. Gov't, P.H.S. 1997;272(48):30122.PubMed
20.
Webster C. Phosphoinositide 3-kinase, but not mitogen-activated protein kinase, pathway is involved in hepatocyte growth factor-mediated protection against bile acid-induced apoptosis in cultured rat hepatocytes. HepatologyJournal Article; Research Support, Non-U.S. Gov’t; Research Support, U.S. Gov’t, P.H.S. 2001;33(3):608–15.PubMed
21.
Fu J, Yang Z, Wei J, Han J, Gu J. Nuclear protein NP60 regulates p38 MAPK activity. J Cell SciJournal Article; Research Support, Non-U.S. Gov’t. 2006;119(Pt 1):115–23.PubMed
22.
Ahmed D, Eide PW, Eilertsen IA, Danielsen SA, Eknæs M, Hektoen M, et al. Epigenetic and genetic features of 24 colon cancer cell lines. Oncogenesis. 2013;2(9):e71.PubMedPubMedCentral
23.
Cottu PH, Muzeau F, Estreicher A, Flejou JF, Iggo R, Thomas G, et al. Inverse correlation between RER+ status and p53 mutation in colorectal cancer cell lines. OncogeneJournal Article; Research Support, Non-U.S. Gov’t. 1996;13(12):2727–30.PubMed
24.
Takenoue T, Kitayama J, Takei Y, Umetani N, Matsuda K, Nita ME, et al. Characterization of dihydropyrimidine dehydrogenase on immunohisto-chemistry in colon carcinoma, and correlation between immunohisto-chemical score and protein level or messenger RNA expression. Ann Oncol. 2000;11(3):273–80.PubMed
25.
Gu C, Wang X, Long T, Wang X, Zhong Y, Ma Y, et al. FSTL1 interacts with VIM and promotes colorectal cancer metastasis via activating the focal adhesion signalling pathway. Cell Death Dis. 2018;9:6.
26.
Bellizzi AM, Frankel WL. Colorectal cancer due to deficiency in DNA mismatch repair function: a review. Adv Anat PatholJournal Article; Review. 2009;16(6):405–17.PubMed
27.
Richman S. Deficient mismatch repair: read all about it (review). Int J OncolJournal Article; Review. 2015;47(4):1189–202.PubMedPubMedCentral
28.
Kane MF, Loda M, Gaida GM, Lipman J, Mishra R, Goldman H, et al. Methylation of the hMLH1 promoter correlates with lack of expression of hMLH1 in sporadic colon tumors and mismatch repair-defective human tumor cell lines. Cancer ResJournal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. 1997;57(5):808–11.PubMed
29.
Fischer PM, Lane DP. Inhibitors of cyclin-dependent kinases as anti-cancer therapeutics. Curr Med Chem. 2000;7(12):1213–45.PubMed
30.
He G, Kuang J, Huang Z, Koomen J, Kobayashi R, Khokhar AR, et al. Upregulation of p27 and its inhibition of CDK2/cyclin E activity following DNA damage by a novel platinum agent are dependent on the expression of p21. Br J CancerJournal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov’t. 2006;95(11):1514–24.PubMedPubMedCentral
31.
Hardwick L, Azzarelli R, Philpott A. Cell cycle-dependent phosphorylation and regulation of cellular differentiation. Biochem Soc TransJournal Article; Review; Research Support, Non-U.S. Gov’t. 2018;46(5):1083–91.PubMed
32.
Chu P, Wu E, Weiss LM. Cytokeratin 7 and cytokeratin 20 expression in epithelial neoplasms: a survey of 435 cases. Mod Pathol. 2000;13(9):962–72.PubMed
33.
Gartel AL, Serfas MS, Gartel M, Goufman E, Wu GS, El-Deiry WS, et al. p21 (WAF1/CIP1) expression is induced in newly nondividing cells in diverse epithelia and during differentiation of the Caco-2 intestinal cell line. Exp Cell ResJournal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. 1996;227(2):171–81.PubMed
34.
Zhao Y, Peng J, Zhang E, Jiang N, Li J, Zhang Q, et al. CD133 expression may be useful as a prognostic indicator in colorectal cancer, a tool for optimizing therapy and supportive evidence for the cancer stem cell hypothesis: a meta-analysis. OncotargetJournal Article; Meta-Analysis; Research Support, Non-U.S. Gov't. 2016;7(9):10023–36.PubMedPubMedCentral
35.
Gawlik-Rzemieniewska N, Bednarek I. The role of NANOG transcriptional factor in the development of malignant phenotype of cancer cells. Cancer Biol TherJournal Article; Research Support, Non-U.S. Gov’t; Review. 2016;17(1):1–10.PubMed
36.
Abbasian M, Mousavi E, Arab-Bafrani Z, Sahebkar A. The most reliable surface marker for the identification of colorectal cancer stem-like cells: a systematic review and meta-analysis. J Cell PhysiolJournal Article; Review. 2018;234(6):8192.PubMed
37.
Pocard M, Bras-Goncalves R, Hamelin R, Northover J, Poupon MF. Response to 5-fluorouracil of orthotopically xenografted human colon cancers with a microsatellite instability: influence of P53 status. Anticancer ResComparative Study; Journal Article; Research Support, Non-U.S. Gov’t. 2000;20(1A):85–90.PubMed
38.
Webber EM, Kauffman TL, Connor EO, Goddard KA. Systematic review of the predictive effect of MSI status in colorectal cancer patients undergoing 5FU-based chemotherapy. BMC Cancer. 2015;15:1.
39.
Tajima A, Hess MT, Cabrera BL, Kolodner RD, Carethers JM. The mismatch repair complex hMutS alpha recognizes 5-fluorouracil-modified DNA: implications for chemosensitivity and resistance. GastroenterologyJournal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. 2004;127(6):1678–84.PubMed
40.
Wang Y, Xia C, Lun Z, Lv Y, Chen W, Li T. Crosstalk between p38 MAPK and caspase-9 regulates mitochondria-mediated apoptosis induced by tetra-alpha-(4-carboxyphenoxy) phthalocyanine zinc photodynamic therapy in LoVo cells. Oncol Rep. 2018;39(1):61–70.PubMed
41.
Wang Y, Xia C, Lv Y, Li C, Mei Q, Li H, et al. Crosstalk influence between P38MAPK and autophagy on mitochondria-mediated apoptosis induced by anti-Fas antibody/Actinomycin D in human hepatoma Bel-7402 cells. Molecules. 2017;22:10.
42.
de la Cruz-Morcillo MA, Valero MLL, Callejas-Valera JL, Arias-González L, Melgar-Rojas P, Galán-Moya EM, et al. P38MAPK is a major determinant of the balance between apoptosis and autophagy triggered by 5-fluorouracil: implication in resistance. Oncogene. 2012;31(9):1073–85.PubMed
43.
Li Q, Lai Z, Yan Z, Peng J, Jin Y, Wei L, et al. Hedyotis diffusa Willd inhibits proliferation and induces apoptosis of 5FU resistant colorectal cancer cells by regulating the PI3K/AKT signaling pathway. Mol Med Rep. 2018;17(1):358–65.PubMed
44.
Lin J, Feng J, Yang H, Yan Z, Li Q, Wei L, et al. Scutellaria barbata D. Don inhibits 5-fluorouracil resistance in colorectal cancer by regulating PI3K/AKT pathway. Oncol Rep. 2017;38(4):2293–300.PubMed
45.
Newmeyer DD, Bossy-Wetzel E, Kluck RM, Wolf BB, Beere HM, Green DR. Bcl-xL does not inhibit the function of Apaf-1. Cell Death DifferJournal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. 2000;7(4):402–7.PubMed
46.
Adams JM, Cory S. Life-or-death decisions by the Bcl-2 protein family. Trends Biochem SciJournal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.; Review. 2001;26(1):61–6.PubMed
47.
Cory S, Adams JM. The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev CancerJournal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov’t, P.H.S.; Review. 2002;2(9):647–56.PubMed
48.
49.
Baretti M, Le DT. DNA mismatch repair in cancer. Pharmacol Ther. 2018;189:45–62.PubMed
50.
de Vos M, Hayward B, Bonthron DT, Sheridan E. Phenotype associated with recessively inherited mutations in DNA mismatch repair (MMR) genes: table 1. Biochem Soc T. 2005;33(4):718–20.
51.
Li GM. Decoding the histone code: role of H3K36me3 in mismatch repair and implications for cancer susceptibility and therapy. Cancer ResJournal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov’t; Review. 2013;73(21):6379–83.PubMed
52.
Li F, Mao G, Tong D, Huang J, Gu L, Yang W, et al. The histone mark H3K36me3 regulates human DNA mismatch repair through its interaction with MutSalpha. CellJournal Article; Research Support, N.I.H., Extramural; Research Support, N.I.H., Intramural; Research Support, Non-U.S. Gov't. 2013;153(3):590–600.PubMedPubMedCentral
53.
Li F, Mao G, Tong D, Huang J, Gu L, Yang W, et al. The histone mark H3K36me3 regulates human DNA mismatch repair through its interaction with MutSα. CELL. 2013;153(3):590–600.PubMedPubMedCentral
54.
Leong V, Lorenowicz J, Kozij N, Guarné A. Nuclear import of human MLH1, PMS2, and MutLα: redundancy is the key. Mol Carcinog. 2009;48(8):742–50.PubMed
55.
Fei J, Ishii H, Hoeksema MA, Meitinger F, Kassavetis GA, Glass CK, et al. NDF, a nucleosome-destabilizing factor that facilitates transcription through nucleosomes. Genes Dev. 2018;32(9–10):682–94.PubMedPubMedCentral
56.
Metadata
Title
Downregulation of GLYR1 contributes to microsatellite instability colorectal cancer by targeting p21 via the p38MAPK and PI3K/AKT pathways
Authors
Zhiyan Hu
Ting Long
Yidan Ma
Jiaxian Zhu
Lingfang Gao
Yan Zhong
Xia Wang
Xiaoyan Wang
Zuguo Li
Publication date
01-12-2020
Publisher
BioMed Central
Published in
Journal of Experimental & Clinical Cancer Research / Issue 1/2020
Electronic ISSN: 1756-9966
DOI
https://doi.org/10.1186/s13046-020-01578-y

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