Podocalyxin is a CD34-related type I transmembrane protein that is highly glycosylated with N-glycan, O-glycan, and keratan sulfate. Podocalyxin was originally found in the podocytes of rat kidney and is reportedly expressed in many types of tumors, including brain tumors, colorectal cancers, and breast cancers. Overexpression of podocalyxin is an independent predictor of progression, metastasis, and poor outcome. We recently immunized mice with recombinant human podocalyxin, which was produced using LN229 glioblastoma cells, and produced a novel antipodocalyxin monoclonal antibody (mAb), PcMab-47, which reacts with endogenous podocalyxin-expressing cancer cell lines and normal cell lines independent of glycosylation in Western blot, flow cytometry, and immunohistochemical analyses. In this study, we performed immunohistochemical analysis against oral cancers using PcMab-47. PcMab-47-stained oral squamous cell carcinoma cells in a cytoplasmic pattern and detected 26/38 (68.4%) of oral squamous cell carcinoma cells on tissue microarrays. These results indicate that PcMab-47 is useful in detecting podocalyxin of oral cancers for immunohistochemical analysis.
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References
1.
McNagnyKM, PetterssonI, RossiF, FlammeI, ShevchenkoA, MannM, and GrafT: Thrombomucin, a novel cell surface protein that defines thrombocytes and multipotent hematopoietic progenitors. J Cell Biol, 1997; 138:1395–1407.
2.
SchopperleWM, KershawDB, and DeWolfWC: Human embryonal carcinoma tumor antigen, Gp200/GCTM-2, is podocalyxin. Biochem Biophys Res Commun, 2003; 300:285–290.
3.
SchopperleWM, and DeWolfWC: The TRA-1-60 and TRA-1-81 human pluripotent stem cell markers are expressed on podocalyxin in embryonal carcinoma. Stem Cells, 2007; 25:723–730.
4.
KerjaschkiD, SharkeyDJ, and FarquharMG: Identification and characterization of podocalyxin—The major sialoprotein of the renal glomerular epithelial cell. J Cell Biol, 1984; 98:1591–1596.
5.
KerjaschkiD, PoczewskiH, DekanG, HorvatR, BalzarE, KraftN, and AtkinsRC: Identification of a major sialoprotein in the glycocalyx of human visceral glomerular epithelial cells. J Clin Invest, 1986; 78:1142–1149.
6.
KershawDB, BeckSG, WharramBL, WigginsJE, GoyalM, ThomasPE, and WigginsRC: Molecular cloning and characterization of human podocalyxin-like protein. Orthologous relationship to rabbit PCLP1 and rat podocalyxin. J Biol Chem, 1997; 272:15708–15714.
HayatsuN, KanekoMK, MishimaK, NishikawaR, MatsutaniM, PriceJE, and KatoY: Podocalyxin expression in malignant astrocytic tumors. Biochem Biophys Res Commun, 2008; 374:394–398.
9.
LarruceaS, ButtaN, RodriguezRB, Alonso-MartinS, Arias-SalgadoEG, AyusoMS, and ParrillaR: Podocalyxin enhances the adherence of cells to platelets. Cell Mol Life Sci, 2007; 64:2965–2974.
10.
LarruceaS, ButtaN, Arias-SalgadoEG, Alonso-MartinS, AyusoMS, and ParrillaR: Expression of podocalyxin enhances the adherence, migration, and intercellular communication of cells. Exp Cell Res, 2008; 314:2004–2015.
11.
TakedaT, McQuistanT, OrlandoRA, and FarquharMG: Loss of glomerular foot processes is associated with uncoupling of podocalyxin from the actin cytoskeleton. J Clin Invest, 2001; 108:289–301.
12.
KangL, YaoC, Khodadadi-JamayranA, XuW, ZhangR, BanerjeeNS, ChangCW, ChowLT, TownesT, and HuK: The universal 3D3 antibody of human PODXL is pluripotent cytotoxic, and identifies a residual population after extended differentiation of pluripotent stem cells. Stem Cells Dev, 2016; 25:556–568.
13.
ToyodaH, NagaiY, KojimaA, and Kinoshita-ToyodaA: Podocalyxin as a major pluripotent marker and novel keratan sulfate proteoglycan in human embryonic and induced pluripotent stem cells. Glycoconj J, 2017; 34:139–145.
SomasiriA, NielsenJS, MakretsovN, McCoyML, PrenticeL, GilksCB, ChiaSK, GelmonKA, KershawDB, HuntsmanDG, McNagnyKM, and RoskelleyCD: Overexpression of the anti-adhesin podocalyxin is an independent predictor of breast cancer progression. Cancer Res, 2004; 64:5068–5073.
16.
SnyderKA, HughesMR, HedbergB, BrandonJ, HernaezDC, BergqvistP, CruzF, PoK, GravesML, TurveyME, NielsenJS, WilkinsJA, McCollSR, BabcookJS, RoskelleyCD, and McNagnyKM: Podocalyxin enhances breast tumor growth and metastasis and is a target for monoclonal antibody therapy. Breast Cancer Res, 2015; 17:46.
17.
GravesML, CipolloneJA, AustinP, BellEM, NielsenJS, GilksCB, McNagnyKM, and RoskelleyCD: The cell surface mucin podocalyxin regulates collective breast tumor budding. Breast Cancer Res, 2016; 18:11.
18.
LarssonA, JohanssonME, WangefjordS, GaberA, NodinB, KucharzewskaP, WelinderC, BeltingM, EberhardJ, JohnssonA, UhlenM, and JirstromK: Overexpression of podocalyxin-like protein is an independent factor of poor prognosis in colorectal cancer. Br J Cancer, 2011; 105:666–672.
19.
KaprioT, FermerC, HagstromJ, MustonenH, BockelmanC, NilssonO, and HaglundC: Podocalyxin is a marker of poor prognosis in colorectal cancer. BMC Cancer, 2014; 14:493.
20.
KaprioT, HagstromJ, FermerC, MustonenH, BockelmanC, NilssonO, and HaglundC: A comparative study of two PODXL antibodies in 840 colorectal cancer patients. BMC Cancer, 2014; 14:494.
21.
LarssonAH, LehnS, WangefjordS, KarneviE, KuteevaE, SundstromM, NodinB, UhlenM, EberhardJ, BirgissonH, and JirstromK: Significant association and synergistic adverse prognostic effect of podocalyxin-like protein and epidermal growth factor receptor expression in colorectal cancer. J Transl Med, 2016; 14:128.
22.
LaitinenA, BockelmanC, HagstromJ, KokkolaA, FermerC, NilssonO, and HaglundC: Podocalyxin as a prognostic marker in gastric cancer. PLoS One, 2015; 10:e0145079.
23.
CaseyG, NevillePJ, LiuX, PlummerSJ, CicekMS, KrumroyLM, CurranAP, McGreevyMR, CatalonaWJ, KleinEA, and WitteJS: Podocalyxin variants and risk of prostate cancer and tumor aggressiveness. Hum Mol Genet, 2006; 15:735–741.
24.
HsuYH, LinWL, HouYT, PuYS, ShunCT, ChenCL, WuYY, ChenJY, ChenTH, and JouTS: Podocalyxin EBP50 ezrin molecular complex enhances the metastatic potential of renal cell carcinoma through recruiting Rac1 guanine nucleotide exchange factor ARHGEF7. Am J Pathol, 2010; 176:3050–3061.
BomanK, LarssonAH, SegerstenU, KuteevaE, JohannessonH, NodinB, EberhardJ, UhlenM, MalmstromPU, and JirstromK: Membranous expression of podocalyxin-like protein is an independent factor of poor prognosis in urothelial bladder cancer. Br J Cancer, 2013; 108:2321–2328.
27.
CipolloneJA, GravesML, KobelM, KallogerSE, PoonT, GilksCB, McNagnyKM, and RoskelleyCD: The anti-adhesive mucin podocalyxin may help initiate the transperitoneal metastasis of high grade serous ovarian carcinoma. Clin Exp Metastasis, 2012; 29:239–252.
28.
HebyM, ElebroJ, NodinB, JirstromK, and EberhardJ: Prognostic and predictive significance of podocalyxin-like protein expression in pancreatic and periampullary adenocarcinoma. BMC Clin Pathol, 2015; 15:10.
29.
SaukkonenK, HagstromJ, MustonenH, JuutiA, NordlingS, FermerC, NilssonO, SeppanenH, and HaglundC: Podocalyxin is a marker of poor prognosis in pancreatic ductal adenocarcinoma. PLoS One, 2015; 10:e0129012.
30.
LinCW, SunMS, and WuHC: Podocalyxin-like 1 is associated with tumor aggressiveness and metastatic gene expression in human oral squamous cell carcinoma. Int J Oncol, 2014; 45:710–718.
31.
BinderZA, SiuIM, EberhartCG, Ap RhysC, BaiRY, StaedtkeV, ZhangH, SmollNR, PiantadosiS, PiccirilloSG, DimecoF, WeingartJD, VescoviA, OliviA, RigginsGJ, and GalliaGL: Podocalyxin-like protein is expressed in glioblastoma multiforme stem-like cells and is associated with poor outcome. PLoS One, 2013; 8:e75945.
32.
AmoL, Tamayo-OrbegozoE, MaruriN, EguizabalC, ZenarruzabeitiaO, RinonM, ArrietaA, SantosS, MongeJ, VesgaMA, BorregoF, and LarruceaS: Involvement of platelet-tumor cell interaction in immune evasion. Potential role of podocalyxin-like protein 1. Front Oncol, 2014; 4:245.
33.
ThomasSN, SchnaarRL, and KonstantopoulosK: Podocalyxin-like protein is an E-/L-selectin ligand on colon carcinoma cells: Comparative biochemical properties of selectin ligands in host and tumor cells. Am J Physiol Cell Physiol, 2009; 296:C505–C513.
34.
DallasMR, ChenSH, StreppelMM, SharmaS, MaitraA, and KonstantopoulosK: Sialofucosylated podocalyxin is a functional E- and L-selectin ligand expressed by metastatic pancreatic cancer cells. Am J Physiol Cell Physiol, 2012; 303:C616–C624.
35.
LaubliH, and BorsigL: Selectins promote tumor metastasis. Semin Cancer Biol, 2010; 20:169–177.
36.
OkiH, KanekoMK, OgasawaraS, TsujimotoY, LiuX, SugawaraM, TakakuboY, TakagiM, and KatoY: Characterization of monoclonal antibody LpMab-7 recognizing non-PLAG domain of podoplanin. Monoclon Antib Immunodiagn Immunother, 2015; 34:174–180.
37.
YamadaS, OgasawaraS, KanekoMK, and KatoY: LpMab-23: A cancer-specific monoclonal antibody against human podoplanin. Monoclon Antib Immunodiagn Immunother, 2017; 36:72–76.
38.
OkiH, OgasawaraS, KanekoMK, TakagiM, YamauchiM, and KatoY: Characterization of monoclonal antibody LpMab-3 recognizing sialylated glycopeptide of podoplanin. Monoclon Antib Immunodiagn Immunother, 2015; 34:44–50.
39.
KanekoMK, OkiH, HozumiY, LiuX, OgasawaraS, TakagiM, GotoK, and KatoY: Monoclonal antibody LpMab-9 recognizes O-glycosylated N-terminus of human podoplanin. Monoclon Antib Immunodiagn Immunother, 2015; 34:310–317.
40.
KatoY, OgasawaraS, OkiH, GoichbergP, HonmaR, FujiiY, and KanekoMK: LpMab-12 established by CasMab technology specifically detects sialylated O-Glycan on Thr52 of platelet aggregation-stimulating domain of human podoplanin. PLoS One, 2016; 11:e0152912.
41.
OgasawaraS, OkiH, KanekoMK, HozumiY, LiuX, HonmaR, FujiiY, NakamuraT, GotoK, TakagiM, and KatoY: Development of monoclonal antibody LpMab-10 recognizing non-glycosylated PLAG1/2 domain including Thr34 of human podoplanin. Monoclon Antib Immunodiagn Immunother, 2015; 34:318–326.
42.
KatoY, OgasawaraS, OkiH, HonmaR, TakagiM, FujiiY, NakamuraT, SaidohN, KannoH, UmetsuM, KamataS, KuboH, YamadaM, SawaY, MoritaK, HaradaH, SuzukiH, and KanekoMK: Novel monoclonal antibody LpMab-17 developed by CasMab technology distinguishes human podoplanin from monkey podoplanin. Monoclon Antib Immunodiagn Immunother, 2016; 35:109–116.
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