Selective assembly of laminin variants by human carcinoma cells.

U M Wewer; E A Wayner; B G Hoffstrom; F Lan; B Meyer-Nielsen; E Engvall; R Albrechtsen

Selective assembly of laminin variants by human carcinoma cells.

U M Wewer, E A Wayner, B G Hoffstrom, F Lan, B Meyer-Nielsen, E Engvall, R Albrechtsen

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

28 Citationer (Scopus)

Abstract

Udgivelsesdato: 1994-Nov

Originalsprog	Engelsk
Tidsskrift	Laboratory Investigation
Vol/bind	71
Udgave nummer	5
Sider (fra-til)	719-30
Antal sider	11
ISSN	0023-6837
Status	Udgivet - 1994

Bibliografisk note

Keywords: Amino Acid Sequence; Animals; Base Sequence; Carcinoma; Cell Adhesion Molecules; Cloning, Molecular; DNA Probes; DNA, Complementary; Gene Expression Regulation, Neoplastic; Humans; Immunologic Techniques; Keratinocytes; Laminin; Male; Mice; Mice, Nude; Molecular Sequence Data; Neoplasm Transplantation; Peptides; RNA, Messenger; Transplantation, Heterologous; Tumor Cells, Cultured

Citationsformater

@article{20ca45905c7c11dd8d9f000ea68e967b,

title = "Selective assembly of laminin variants by human carcinoma cells.",

abstract = "BACKGROUND: The laminins are heterotrimeric basement membrane glycoproteins. Eight subunits that can be assembled into laminins have been characterized and are known as: A, B1, B2, S, M, K, B2t, B1k laminin chains. Although many neoplastic cells secrete laminins and some of them even assemble basement membranes, the pattern of production of various laminin subunits remains to be explored. EXPERIMENTAL DESIGN: The expression of laminin was examined in several human carcinoma cells using a panel of specific cDNA probes as well as polyclonal and chain specific monoclonal antibodies. For this purpose a human laminin S chain 2 kb cDNA was isolated and characterized and used together with existing probes for laminin chains. RESULTS: All carcinoma cell lines had a high level of expression of three light chains (B1, S and B2) mRNA. In contrast, the heavy chains of laminin, A and M, were expressed in negligible amounts as detected by Northern blotting and PCR. The only exception was the HU-1 lung adenocarcinoma cell line which expressed significant quantities of laminin M chain mRNA and lower levels of laminin A chain mRNA. The presence in the HU-1 cells of translated polypeptides was demonstrated by immunofluorescence staining. The cells contained both B1 and S chain laminin in the cell layer, but preferentially secreted the B1 chain into the culture supernatant as shown by Western blotting. The 300 to 400 kDa M chain immunoreactive band was found in laminin secreted into the culture medium of HU-1 cells. Immunoprecipitation of biosynthetically labeled proteins showed that the M chain was synthesized as a complex with B chains. Little or no A chain laminin was detected in the culture medium supernatant. HU-1 cells also synthesized the newly described laminin variant, epiligrin which was secreted into the medium. Thus, the HU-1 cells secreted two laminin variants: M-B1-B12 laminin and epiligrin into the culture medium. Immunostaining of HU-1 nude mice tumors showed that tumor basement membranes contained M, B1, and B2 laminin and epiligrin immunoreactivity but apparently no S chain. CONCLUSIONS: All human carcinoma cell lines produced laminin chains B1, B2 and S, but no or little A or M. The only exception was the lung carcinoma cell line HU-1. Human HU-1 carcinoma cells in culture synthesize several homologous laminin chains and regulate the process of assembly, secretion and deposition of laminin variants into tumor basement membranes. These data indicate that the tumor cells vary among themselves with regards to laminin production and that some of them, like HU-1 may produce essentially all laminin chains simultaneously.",

author = "Wewer, {U M} and Wayner, {E A} and Hoffstrom, {B G} and F Lan and B Meyer-Nielsen and E Engvall and R Albrechtsen",

note = "Keywords: Amino Acid Sequence; Animals; Base Sequence; Carcinoma; Cell Adhesion Molecules; Cloning, Molecular; DNA Probes; DNA, Complementary; Gene Expression Regulation, Neoplastic; Humans; Immunologic Techniques; Keratinocytes; Laminin; Male; Mice; Mice, Nude; Molecular Sequence Data; Neoplasm Transplantation; Peptides; RNA, Messenger; Transplantation, Heterologous; Tumor Cells, Cultured",

year = "1994",

language = "English",

volume = "71",

pages = "719--30",

journal = "Laboratory Investigation",

issn = "0023-6837",

publisher = "nature publishing group",

number = "5",

}

TY - JOUR

T1 - Selective assembly of laminin variants by human carcinoma cells.

AU - Wewer, U M

AU - Wayner, E A

AU - Hoffstrom, B G

AU - Lan, F

AU - Meyer-Nielsen, B

AU - Engvall, E

AU - Albrechtsen, R

N1 - Keywords: Amino Acid Sequence; Animals; Base Sequence; Carcinoma; Cell Adhesion Molecules; Cloning, Molecular; DNA Probes; DNA, Complementary; Gene Expression Regulation, Neoplastic; Humans; Immunologic Techniques; Keratinocytes; Laminin; Male; Mice; Mice, Nude; Molecular Sequence Data; Neoplasm Transplantation; Peptides; RNA, Messenger; Transplantation, Heterologous; Tumor Cells, Cultured

PY - 1994

Y1 - 1994

N2 - BACKGROUND: The laminins are heterotrimeric basement membrane glycoproteins. Eight subunits that can be assembled into laminins have been characterized and are known as: A, B1, B2, S, M, K, B2t, B1k laminin chains. Although many neoplastic cells secrete laminins and some of them even assemble basement membranes, the pattern of production of various laminin subunits remains to be explored. EXPERIMENTAL DESIGN: The expression of laminin was examined in several human carcinoma cells using a panel of specific cDNA probes as well as polyclonal and chain specific monoclonal antibodies. For this purpose a human laminin S chain 2 kb cDNA was isolated and characterized and used together with existing probes for laminin chains. RESULTS: All carcinoma cell lines had a high level of expression of three light chains (B1, S and B2) mRNA. In contrast, the heavy chains of laminin, A and M, were expressed in negligible amounts as detected by Northern blotting and PCR. The only exception was the HU-1 lung adenocarcinoma cell line which expressed significant quantities of laminin M chain mRNA and lower levels of laminin A chain mRNA. The presence in the HU-1 cells of translated polypeptides was demonstrated by immunofluorescence staining. The cells contained both B1 and S chain laminin in the cell layer, but preferentially secreted the B1 chain into the culture supernatant as shown by Western blotting. The 300 to 400 kDa M chain immunoreactive band was found in laminin secreted into the culture medium of HU-1 cells. Immunoprecipitation of biosynthetically labeled proteins showed that the M chain was synthesized as a complex with B chains. Little or no A chain laminin was detected in the culture medium supernatant. HU-1 cells also synthesized the newly described laminin variant, epiligrin which was secreted into the medium. Thus, the HU-1 cells secreted two laminin variants: M-B1-B12 laminin and epiligrin into the culture medium. Immunostaining of HU-1 nude mice tumors showed that tumor basement membranes contained M, B1, and B2 laminin and epiligrin immunoreactivity but apparently no S chain. CONCLUSIONS: All human carcinoma cell lines produced laminin chains B1, B2 and S, but no or little A or M. The only exception was the lung carcinoma cell line HU-1. Human HU-1 carcinoma cells in culture synthesize several homologous laminin chains and regulate the process of assembly, secretion and deposition of laminin variants into tumor basement membranes. These data indicate that the tumor cells vary among themselves with regards to laminin production and that some of them, like HU-1 may produce essentially all laminin chains simultaneously.

AB - BACKGROUND: The laminins are heterotrimeric basement membrane glycoproteins. Eight subunits that can be assembled into laminins have been characterized and are known as: A, B1, B2, S, M, K, B2t, B1k laminin chains. Although many neoplastic cells secrete laminins and some of them even assemble basement membranes, the pattern of production of various laminin subunits remains to be explored. EXPERIMENTAL DESIGN: The expression of laminin was examined in several human carcinoma cells using a panel of specific cDNA probes as well as polyclonal and chain specific monoclonal antibodies. For this purpose a human laminin S chain 2 kb cDNA was isolated and characterized and used together with existing probes for laminin chains. RESULTS: All carcinoma cell lines had a high level of expression of three light chains (B1, S and B2) mRNA. In contrast, the heavy chains of laminin, A and M, were expressed in negligible amounts as detected by Northern blotting and PCR. The only exception was the HU-1 lung adenocarcinoma cell line which expressed significant quantities of laminin M chain mRNA and lower levels of laminin A chain mRNA. The presence in the HU-1 cells of translated polypeptides was demonstrated by immunofluorescence staining. The cells contained both B1 and S chain laminin in the cell layer, but preferentially secreted the B1 chain into the culture supernatant as shown by Western blotting. The 300 to 400 kDa M chain immunoreactive band was found in laminin secreted into the culture medium of HU-1 cells. Immunoprecipitation of biosynthetically labeled proteins showed that the M chain was synthesized as a complex with B chains. Little or no A chain laminin was detected in the culture medium supernatant. HU-1 cells also synthesized the newly described laminin variant, epiligrin which was secreted into the medium. Thus, the HU-1 cells secreted two laminin variants: M-B1-B12 laminin and epiligrin into the culture medium. Immunostaining of HU-1 nude mice tumors showed that tumor basement membranes contained M, B1, and B2 laminin and epiligrin immunoreactivity but apparently no S chain. CONCLUSIONS: All human carcinoma cell lines produced laminin chains B1, B2 and S, but no or little A or M. The only exception was the lung carcinoma cell line HU-1. Human HU-1 carcinoma cells in culture synthesize several homologous laminin chains and regulate the process of assembly, secretion and deposition of laminin variants into tumor basement membranes. These data indicate that the tumor cells vary among themselves with regards to laminin production and that some of them, like HU-1 may produce essentially all laminin chains simultaneously.

M3 - Journal article

C2 - 7967523

SN - 0023-6837

VL - 71

SP - 719

EP - 730

JO - Laboratory Investigation

JF - Laboratory Investigation

IS - 5

ER -