Glycosylphosphatidylinositol Anchor Modification Machinery Deficiency Is Responsible for the Formation of Pro-Prion Protein (PrP) in BxPC-3 Protein and Increases Cancer Cell Motility

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2015 Dec 20

PMID 26683373

Citations 9

Authors

Liheng Yang

Zhenxing Gao

Lipeng Hu

Guiru Wu

Xiaowen Yang

Lihua Zhang

Ying Zhu

Boon-Seng Wong

Wei Xin

Man-Sun Sy

Chaoyang Li

Affiliations

Soon will be listed here.

Abstract

The normal cellular prion protein (PrP) is a glycosylphosphatidylinositol (GPI)-anchored cell surface glycoprotein. However, in pancreatic ductal adenocarcinoma cell lines, such as BxPC-3, PrP exists as a pro-PrP retaining its glycosylphosphatidylinositol (GPI) peptide signaling sequence. Here, we report the identification of another pancreatic ductal adenocarcinoma cell line, AsPC-1, which expresses a mature GPI-anchored PrP. Comparison of the 24 genes involved in the GPI anchor modification pathway between AsPC-1 and BxPC-3 revealed 15 of the 24 genes, including PGAP1 and PIG-F, were down-regulated in the latter cells. We also identified six missense mutations in DPM2, PIG-C, PIG-N, and PIG-P alongside eight silent mutations. When BxPC-3 cells were fused with Chinese hamster ovary (CHO) cells, which lack endogenous PrP, pro-PrP was successfully converted into mature GPI-anchored PrP. Expression of the individual gene, such as PGAP1, PIG-F, or PIG-C, into BxPC-3 cells does not result in phosphoinositide-specific phospholipase C sensitivity of PrP. However, when PIG-F but not PIG-P is expressed in PGAP1-expressing BxPC-3 cells, PrP on the surface of the cells becomes phosphoinositide-specific phospholipase C-sensitive. Thus, low expression of PIG-F and PGAP1 is the major factor contributing to the accumulation of pro-PrP. More importantly, BxPC-3 cells expressing GPI-anchored PrP migrate much slower than BxPC-3 cells bearing pro-PrP. In addition, GPI-anchored PrP-bearing AsPC-1 cells also migrate slower than pro-PrP bearing BxPC-3 cells, although both cells express filamin A. "Knocking out" PRNP in BxPC-3 cell drastically reduces its migration. Collectively, these results show that multiple gene irregularity in BxPC-3 cells is responsible for the formation of pro-PrP, and binding of pro-PrP to filamin A contributes to enhanced tumor cell motility.

Citing Articles

The expression of pro-prion, a transmembrane isoform of the prion protein, leads to the constitutive activation of the canonical Wnt/β-catenin pathway to sustain the stem-like phenotype of human glioblastoma cells.

Corsaro A, Dellacasagrande I, Tomanelli M, Pagano A, Barbieri F, Thellung S Cancer Cell Int. 2024; 24(1):426.

PMID: 39716276 PMC: 11667964. DOI: 10.1186/s12935-024-03581-1.

Integration of metabolites from meta-analysis with transcriptome reveals enhanced SPHK1 in PDAC with a background of pancreatitis.

Ketavarapu V, Ravikanth V, Sasikala M, Rao G, Devi C, Sripadi P BMC Cancer. 2022; 22(1):792.

PMID: 35854233 PMC: 9295503. DOI: 10.1186/s12885-022-09816-6.

Upregulation of phosphatidylinositol glycan anchor biosynthesis class C is associated with unfavorable survival prognosis in patients with hepatocellular carcinoma.

Peng X, Lei C, He A, Luo R, Cai Y, Dong W Oncol Lett. 2021; 21(3):237.

PMID: 33613726 PMC: 7856693. DOI: 10.3892/ol.2021.12498.

The Role of Cellular Prion Protein in Promoting Stemness and Differentiation in Cancer.

Ryskalin L, Biagioni F, Busceti C, Giambelluca M, Morelli L, Frati A Cancers (Basel). 2021; 13(2).

PMID: 33418999 PMC: 7825291. DOI: 10.3390/cancers13020170.

Prion Protein at the Leading Edge: Its Role in Cell Motility.

Prado M, Escobar M, Alves R, Coelho B, Fernandes C, Boccacino J Int J Mol Sci. 2020; 21(18).

PMID: 32932634 PMC: 7555277. DOI: 10.3390/ijms21186677.

References

Stossel T, Condeelis J, Cooley L, Hartwig J, Noegel A, Schleicher M . Filamins as integrators of cell mechanics and signalling. Nat Rev Mol Cell Biol. 2001; 2(2):138-45. DOI: 10.1038/35052082. View

Yang L, Zhang Y, Hu L, Zhu Y, Sy M, Li C . A panel of monoclonal antibodies against the prion protein proves that there is no prion protein in human pancreatic ductal epithelial cells. Virol Sin. 2014; 29(4):228-36. PMC: 8206262. DOI: 10.1007/s12250-014-3480-4. View

Stossel T, Hartwig J . Filling gaps in signaling to actin cytoskeletal remodeling. Dev Cell. 2003; 4(4):444-5. DOI: 10.1016/s1534-5807(03)00098-4. View

Takeda J, Miyata T, Kawagoe K, Iida Y, Endo Y, Fujita T . Deficiency of the GPI anchor caused by a somatic mutation of the PIG-A gene in paroxysmal nocturnal hemoglobinuria. Cell. 1993; 73(4):703-11. DOI: 10.1016/0092-8674(93)90250-t. View

Moss J, Balducci E, CAVANAUGH E, Kim H, Konczalik P, Lesma E . Characterization of NAD:arginine ADP-ribosyltransferases. Mol Cell Biochem. 1999; 193(1-2):109-13. View

Ho J, Cheung S, Patil M, Chen X, Fan S . Increased expression of glycosyl-phosphatidylinositol anchor attachment protein 1 (GPAA1) is associated with gene amplification in hepatocellular carcinoma. Int J Cancer. 2006; 119(6):1330-7. DOI: 10.1002/ijc.22005. View

Pan Y, Zhao L, Liang J, Liu J, Shi Y, Liu N . Cellular prion protein promotes invasion and metastasis of gastric cancer. FASEB J. 2006; 20(11):1886-8. DOI: 10.1096/fj.06-6138fje. View

Maeda Y, Ashida H, Kinoshita T . CHO glycosylation mutants: GPI anchor. Methods Enzymol. 2006; 416:182-205. DOI: 10.1016/S0076-6879(06)16012-7. View

Richt J, Kasinathan P, Hamir A, Castilla J, Sathiyaseelan T, Vargas F . Production of cattle lacking prion protein. Nat Biotechnol. 2007; 25(1):132-8. PMC: 2813193. DOI: 10.1038/nbt1271. View

10.

Meslin F, Conforti R, Mazouni C, Morel N, Tomasic G, Drusch F . Efficacy of adjuvant chemotherapy according to Prion protein expression in patients with estrogen receptor-negative breast cancer. Ann Oncol. 2007; 18(11):1793-8. DOI: 10.1093/annonc/mdm406. View

11.

Zhou L, Shang Y, Liu C, Li J, Hu H, Liang C . Overexpression of PrPc, combined with MGr1-Ag/37LRP, is predictive of poor prognosis in gastric cancer. Int J Cancer. 2014; 135(10):2329-37. PMC: 4277329. DOI: 10.1002/ijc.28883. View

12.

Prusiner S, Groth D, Serban A, Koehler R, Foster D, Torchia M . Ablation of the prion protein (PrP) gene in mice prevents scrapie and facilitates production of anti-PrP antibodies. Proc Natl Acad Sci U S A. 1993; 90(22):10608-12. PMC: 47826. DOI: 10.1073/pnas.90.22.10608. View

13.

Sailer A, Bueler H, Fischer M, Aguzzi A, Weissmann C . No propagation of prions in mice devoid of PrP. Cell. 1994; 77(7):967-8. DOI: 10.1016/0092-8674(94)90436-7. View

14.

Lehmann S, Harris D . A mutant prion protein displays an aberrant membrane association when expressed in cultured cells. J Biol Chem. 1995; 270(41):24589-97. DOI: 10.1074/jbc.270.41.24589. View

15.

Brown D, Qin K, Herms J, Madlung A, Manson J, Strome R . The cellular prion protein binds copper in vivo. Nature. 1997; 390(6661):684-7. DOI: 10.1038/37783. View

16.

Diep D, Nelson K, Raja S, Pleshak E, Buckley J . Glycosylphosphatidylinositol anchors of membrane glycoproteins are binding determinants for the channel-forming toxin aerolysin. J Biol Chem. 1998; 273(4):2355-60. DOI: 10.1074/jbc.273.4.2355. View

17.

Zanusso G, Liu D, Ferrari S, Hegyi I, Yin X, Aguzzi A . Prion protein expression in different species: analysis with a panel of new mAbs. Proc Natl Acad Sci U S A. 1998; 95(15):8812-6. PMC: 21159. DOI: 10.1073/pnas.95.15.8812. View

18.

Pauly P, Harris D . Copper stimulates endocytosis of the prion protein. J Biol Chem. 1998; 273(50):33107-10. DOI: 10.1074/jbc.273.50.33107. View

19.

Chen R, Knez J, Merrick W, Medof M . Comparative efficiencies of C-terminal signals of native glycophosphatidylinositol (GPI)-anchored proproteins in conferring GPI-anchoring. J Cell Biochem. 2001; 84(1):68-83. DOI: 10.1002/jcb.1267. View

20.

Hong Y, Maeda Y, Watanabe R, Inoue N, Ohishi K, Kinoshita T . Requirement of PIG-F and PIG-O for transferring phosphoethanolamine to the third mannose in glycosylphosphatidylinositol. J Biol Chem. 2000; 275(27):20911-9. DOI: 10.1074/jbc.M001913200. View