Expression of the type-1 repeats of thrombospondin-1 inhibits tumor growth through activation of transforming growth factor-beta.
In the present study, the type-1 repeats of thrombospondin-1 (TSP-1) were transfected into A431 cells. Expression of all three type-1 repeats (3TSR) and expression of just the second type-1 repeat containing the transforming growth factor (TGF)-beta activating sequence KRFK (TSR2 + KRFK) significantly inhibited in vivo tumor angiogenesis and growth in nude mice. These tumors expressed increased levels of both active and total TGF-beta. A431 cells expressing the second type-1 repeat without the KRFK sequence (TSR2 - KRFK) produced tumors that were slightly larger than the 3TSR and TSR2 + KRFK tumors. These tumors expressed elevated levels of active TGF-beta but levels of total TGF-beta were not different from control tumors. Injection of the peptide, LSKL, which blocks TSP-1 activation of TGF-beta, reversed the growth inhibition observed with cells expressing TSR2 + KRFK to a level comparable to controls. Various residues in the WSHWSPW region and the VTCG sequence of both TSR2+/- KRFK were mutated. Although mutation of the VTCG sequence had no significant effect on tumor growth, mutation of the WSHWSPW sequence reduced inhibition of tumor growth. These findings suggest that the inhibition of tumor angiogenesis and growth by endogenous TSP-1 involves regulation of both active and total TGF-beta and the sequences KRFK and WSHWSPW in the second type-1 repeat.
Department of Pathology, Beth Israel Deaconess Medical Center, 330 Brookline Ave., Research North 270C, Boston MA 02215, USA.
Karen O
KO
Yee
Michael
M
Streit
Thomas
T
Hawighorst
Michael
M
Detmar
Jack
J
Lawler
D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000098:blood supply / Q000473:pathology*; D015870:Gene Expression / Q000502:physiology*; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008175:Lung Neoplasms / Q000098:blood supply / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009154:Mutation; D009389:Neovascularization, Pathologic / Q000517:prevention & control; D010446:Peptide Fragments / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D015996:Survival Rate; D019700:Thrombospondin 1 / Q000235:genetics*; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D020533:Angiogenesis Inhibitors; D010446:Peptide Fragments; D019700:Thrombospondin 1; D016212:Transforming Growth Factor beta; D008156:Luciferases