Studies of human cell hybrids have provided evidence that the tumorigenicity of a cervical carcinoma (HeLa) is under the control of a putative tumor suppressor on chromosome 11. Using these human cell hybrids, we found a tumor-associated glycosylation change in the glucose transporter GLUT1, which is an N-linked glycoprotein at the plasma membrane. The non-tumorigenic HeLa × fibroblast cell hybrid CGL1 and the normal diploid fibroblast WI38 expressed the 50–55 kDa GLUT1, whereas in a tumorigenic segregant hybrid, CGL4, as well as in parental HeLa cells, GLUT1 glycosylation was altered and its molecular mass was about 70 kDa. However, the altered GLUT1 glycosylation was not observed in SV40-transformed WI38 cells, suggesting a correlation between this glycosylation change and a putative tumor suppressor function. Further investigations using glycosidases, glycosylation inhibitors and lectin-affinity chromatography demonstrated that the tumor-associated glycosylation change in GLUT1 was mainly due to the increase in N-acetyl-lactosamine repeats in the N-linked oligosaccharides. In accordance with the altered glycosylation, affinity for 2-deoxyglucose in the tumorigenic CGL4 cells increased 2-fold, but there was little change in the Vmax. These results suggest there may be a functional role for the modulation by glycosylation of GLUT1 in the tumorigenic behavior of CGL4 and HeLa cells.

REFERENCES

Asano
T.
,
Katagiri
H.
,
Takata
K.
,
Lin
J.-L.
,
Ishihara
H.
,
Inukai
K.
,
Tsukuda
K.
,
Kikuchi
M.
,
Hirano
H.
,
Yazaki
Y.
,
Oka
Y.
(
1991
).
The role of N -glycosylation of GLUT1 for glucose transport activity.
J. Biol. Chem
266
,
24632
24636
Bell
G. I.
,
Burant
C. F.
,
Takeda
J.
,
Gould
G. M.
(
1993
).
Structure and function of mammalian facilitative sugar transporters.
J. Biol. Chem
268
,
19161
19164
Birnbaum
M. J.
,
Haspel
H. C.
,
Rosen
O. M.
(
1986
).
Cloning and characterization of a cDNA encoding the rat brain glucose-transporter protein.
Proc. Nat. Acad. Sci. USA
83
,
5784
5788
Birnbaum
M. J.
,
Haspel
H. C.
,
Rosen
O. M.
(
1987
).
Transformation of rat fibroblasts by FSV rapidly increases glucose transporter gene transcription.
Science
235
,
1495
1498
Bramwell
M. E.
,
Davies
A.
,
Baldwin
S. A.
(
1990
).
Heterogeniety of the glucose transporter in malignant and suppressed hybrid cells.
Exp. Cell Res
188
,
97
104
Buck
C. A.
,
Fuhrer
J. P.
,
Soslau
G.
,
Warren
L.
(
1974
).
Membrane glycopeptides from subcellular fractions of control and virus-transformed cells.
J. Biol. Chem
249
,
1541
1550
Charron
M. J.
,
Brosius
F. C.
III
,
Alper
S. L.
,
Lodish
H. F.
(
1989
).
A glucose transport protein expressed predominately in insulin-responsive tissues.
Proc. Nat. Acad. Sci. USA
86
,
2535
2539
Cummings
R. D.
,
Kornfeld
S.
(
1984
).
The distribution of repeating [Gal1,4GlcNAc 1,3] sequences in asparagine-linked oligosaccharides of the mouse lymphoma cell lines BW5147 and PHAR2.1.
J. Biol. Chem
259
,
6253
6260
Dennis
J. W.
,
Laferte
S.
,
Waghorne
C.
,
Breitman
M. L.
,
Kerbel
R. S.
(
1987
).
1-6 Branching of Asn-linked oligosaccharides is directly associated with metastasis.
Science
236
,
582
585
Der
C. J.
,
Stanbridge
E. J.
(
1981
).
A tumor-specific membrane phosphoprotein marker in human cell hybrids.
Cell
26
,
429
438
Endo
T.
,
Kasahara
M.
,
Kobata
A.
(
1990
).
Structures of the asparagine-linked sugar chain of glucose transporter from human erythrocytes.
Biochemistry
29
,
9126
9134
Fearon
E. R.
,
Vogelstein
B.
(
1990
).
A genetic model for colorectal tumorigenesis.
Cell
61
,
759
767
Flier
J. S.
,
Mueckler
M. M.
,
Usher
P.
,
Lodish
H. F.
(
1987
).
Elevated levels of glucose transport and transporter mRNA are induced by ras or src oncogenes.
Science
235
,
1492
1495
Girardi
A. J.
,
Weinstein
D.
,
Moorhead
P. S.
(
1966
).
SV40 transformation of human diploid cells.
Ann. Med. Exp. Fenn
44
,
242
254
Gross
V.
,
Tran-Thi
T.-A.
,
Vosbeck
K.
,
Heinrich
P. C.
(
1983
).
Effect of swainsonine on the processing of the asparagine-linked carbohydrate chains of1-antitrypsin in rat hepatocytes.
J. Biol. Chem
258
,
4032
4036
Hampton
G. M.
,
Penny
L. A.
,
Baergen
R. N.
,
Larson
A.
,
Brewer
C.
,
Liao
S.
,
Busby-Earle
R. M. C.
,
Williams
A. W. R.
,
Steel
C. M.
,
Bird
C. C.
,
Stanbridge
E. J.
,
Evans
G. A.
(
1994
).
Loss of heterozygosity in cervical carcinoma: subchromosomal localization of a putative tumor-suppressor gene to chromosome 11q22-q24.
Proc. Nat. Acad. Sci. USA
91
,
6953
6957
Harris
H.
(
1988
).
The analysis of malignancy by cell fusion: the position in 1988.
Cancer Res
48
,
3302
3306
Hiraki
Y.
,
Rosen
O. M.
,
Birnbaum
M. J.
(
1988
).
Growth factors rapidly induce expression of the glucose transporter gene.
J. Biol. Chem
263
,
13655
13662
Hiraki
Y.
,
Garcia de Herreros
A.
,
Birnbaum
M. J.
(
1989
).
Transformation stimulates glucose transporter gene expression in the absence of protein kinase C.
Proc. Nat. Acad. Sci. USA
86
,
8252
8256
Haspel
H. C.
,
Rosenfeld
M.G.
,
Rosen
O. M.
(
1988
).
Characterization of antisera to a synthetic carboxyl-terminal peptide of the glucose transporter protein.
J. Biol. Chem
263
,
398
403
Haspel
H. C.
,
Birnbaum
M. J.
,
Wilk
E. W.
,
Rosen
O. M.
(
1985
).
Biosynthetic precursors and in vitro translation products of the glucose transporter of human hepatocarcinoma cells, human fibroblasts, and murine preadipocytes.
J. Biol. Chem
260
,
7219
7225
Hatanaka
M.
(
1974
).
Transport of sugars in tumor cell membranes.
Biochim. Biophys. Acta
355
,
77
104
Hunter
T.
,
Pines
J.
(
1994
).
Cyclins and cancer II: cyclin D and CDK inhibitors come of age.
Cell
79
,
573
582
Kaestner
K. H.
,
Christy
R. J.
,
McLenithan
J. C.
,
Braiterman
L. T.
,
Cornelius
P.
,
Pekala
P. H.
,
Lane
M. D.
(
1989
).
Sequence, tissue distribution, and differential expression of mRNA for a putative insulin-responsive glucose transporter in mouse 3T3-L1 adipocytes.
Proc. Nat. Acad. Sci. USA
86
,
3150
3154
Kitagawa
T.
,
Tanaka
M.
,
Akamatsu
Y.
(
1989
).
Regulation of glucose transport activity and expression of glucose transporter mRNA by serum, growth factors and phorbol ester in quiescent mouse fibroblasts.
Biochim. Biophys. Acta
980
,
100
108
Kitagawa
T.
,
Masumi
A.
,
Akamatsu
Y.
(
1991
).
Transforming growth factor-1 stimulates glucose uptake and the expression of glucose transporter mRNA in quiescent mouse 3T3 cells.
J. Biol. Chem
266
,
18066
18071
Kornfeld
R.
,
Kornfeld
S.
(
1985
).
Assembly of asparagine-linked oligosaccharides.
Annu. Rev. Biochem
54
,
631
664
Latham
K. M.
,
Stanbridge
E. J.
(
1990
).
Identification of the HeLa tumor-associated antigen, p75/150, as intestinal alkaline phosphatase and evidence for its transcriptional regulation.
Proc. Nat. Acad. Sci. USA
87
,
1263
1267
Latham
K. M.
,
Stanbridge
E. J.
(
1992
).
Examination of the oncogenic potential of a tumor-associated antigen, intestinal alkaline phospatase, in HeLafibroblast cell hybrids.
Cancer Res
52
,
616
622
Marshall
C. J.
(
1991
).
Tumor suppressor genes.
Cell
64
,
313
326
Massague
J.
(
1990
).
The transforming growth factor-family.
Annu. Rev. Cell Biol
6
,
597
641
Masumi
A.
,
Akamatsu
Y.
,
Kitagawa
T.
(
1993
).
Modulation of the synthesis and glycosylation of the glucose transporter protein by transforming growth factor-1 in Swiss 3T3 fibroblasts.
Biochim. Biophys. Acta
1145
,
227
234
Masumi
A.
,
Akamatsu.
Y.
,
Kitagawa
T.
(
1994
).
Alteration by transforming growth factor-1 of asparagine-linked sugar chains in glucose transporter protein in Swiss 3T3 cells.
Biochim. Biophys. Acta
1221
,
330
338
Rademacher
T. W.
,
Parekh
R. B.
,
Dwek
R. A.
(
1988
).
Glycobiology.
Annu. Rev. Biochem
57
,
785
838
Rhoads
D. B.
,
Takano
M.
,
Gattoni-Celli
S.
,
Chen
C.-C.
,
Isselbacher
K. J.
(
1988
).
Evidence for expression of the facilitated glucose transporter in rat hepatocytes.
Proc. Nat. Acad. Sci. USA
85
,
9042
9046
Saxon
P. J.
,
Srivatsan
E. S.
,
Stanbridge
E. J.
(
1986
).
Introduction of human chromosome 11 via microcell transfer controls tumorigenic expression of HeLa cells.
EMBO J
5
,
3461
3466
Schroer
D. W.
,
Frost
S. C.
,
Kohanski
R. A.
,
Lane
M. D.
,
Lienhard
G. E.
(
1986
).
Identification and partial purification of the insulin-responsive glucose transporter from 3T3-L1 adipocytes.
Biochim. Biophys. Acta
885
,
317
326
Sherr
C. J.
(
1994
).
G1 phase progression: cycling on cue.
Cell
79
,
551
555
Sogin
D.C.
,
Hinkle
P.C.
(
1980
).
Immunological identification of the human erythrocyte glucose transporter.
Proc. Nat. Acad. Sci. USA
77
,
5725
5729
Stanbridge
E. J.
(
1976
).
Suppression of malignancy in human cells.
Nature
260
,
17
20
Stanbridge
E. J.
,
Wilkinson
J.
(
1978
).
Analysis of malignancy in human cells: malignant and transformed phenotypes are under separate genetic control.
Proc. Nat. Acad. Sci. USA
75
,
1466
1469
Stanbridge
E. J.
,
Flandermeyer
R. R.
,
Daniels
D. W.
,
Nelson-Rees
W. A.
(
1981
).
Specific chromosome loss associated with the expression of tumorigenicity in human cell hybrids.
Somat. Cell Genet
7
,
699
712
Stanbridge
E. J.
,
Der
C. J.
,
Doersen
C.-J.
,
Nishimi
R. Y.
,
Peehl
D. M.
,
Weissman
B. E.
,
Wilkinson
J. E.
(
1982
).
Human cell hybrids: analysis of transformation and tumorigenicity.
Science
215
,
252
259
Tarentino
A. L.
,
Gomez
C. M.
,
Plummer
T. H.
Jr.
(
1985
).
Deglycosylation of asparagine-linked glycans by peptide: N -glycosidase F.
Biochemistry
24
,
4665
4671
Weinberg
R. A.
(
1991
).
Tumor suppressor genes.
Science
254
,
1138
1146
White
M. K.
,
Bramwell
M. E.
,
Harris
H.
(
1981
).
Hexose transport in hybrids between malignant and normal cells.
Nature
294
,
232
235
White
M. K.
,
Bramwell
M. E.
,
Harris
H.
(
1983
).
Kinetic parameters of hexose transport in hybrid between malignant and non-malignant cells.
J. Cell Sci
62
,
49
80
Yamamoto
T.
,
Seino
Y.
,
Fukumoto
H.
,
Koh
G.
,
Yano
H.
,
Inagaki
N.
,
Yamada
Y.
,
Inoue
K.
,
Manabe
T.
,
Imura
H.
(
1990
).
Over-expression of facilitative glucose transporter genes in human cancer.
Biochem. Biophys. Res. Commun
170
,
223
230
Yamashita
K.
,
Tachibana
Y.
,
Ohkura
T.
,
Kobata
A.
(
1985
).
Enzymatic basis for the structural changes of asparagine-linked sugar chains of membrane glycoproteins of baby hamster kidney cells induced by polyoma transformation.
J. Biol. Chem
260
,
3963
3969
Yamashita
K.
,
Totani
K.
,
Ohkura
T.
,
Takahashi
S.
,
Goldstein
I. J.
,
Kobata
A.
(
1987
).
Carbohydrate binding properties of complex-type oligosaccharides on immobilized Datura stramonium lectin.
J. Biol. Chem
262
,
1602
1607
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