LRRC4 inhibits glioblastoma cell proliferation, migration, and angiogenesis by downregulating pleiotropic cytokine expression and responses.
Leucine-rich repeat C4 (LRRC4) has been shown to inhibit glioma cell proliferation, however, little is known about the mechanism(s) underlying the action of LRRC4. Here, we show that two glioblstoma U251 cell clones stably expressing LRRC4 were established. LRRC4 expression significantly inhibited the expression of some cytokines and their receptors determined by microarray and Western blot assays, and dramatically reduced cytokine-induced AP-1, NF-kB, and CyclinD1 activation in glioma cells. Furthermore, LRRC4 expression in glioma cells significantly downregulated spontaneous and cytokine-induced expression of K-RAS and phosphorylation of c-Raf, ERK, AKT, NF-kBp65, p70S6K, and PKC, suggesting that LRRC4 inhibited receptor tyrosine kinase (RTK) signaling pathways. Moreover, treatment with bFGF, IGF1, or IGF2 stimulated LRRC4(-/-), but not the LRRC4(+), glioma cell proliferation, indicating that LRRC4 mitigated cytokine-stimulated proliferation in glioma cells. In addition, treatment of LRRC4(-/-) glioma cells with EGF, IGF2, or PDGF promoted long distance mobilization, but induced little migration in LRRC4(+) glioma cells, suggesting that LRRC4 retarded cytokine-promoted glioma cell migration in vitro. Finally, human vessel endothelial cells (ECV304) treated with VEGF grew, aligned and formed hollow tube-like structures in vitro. In contrast, LRRC4(+) ECV304 treated with VEGF failed to form vessel-tube structures. Collectively, LRRC4 expression inhibited the expression of some growth factors, cytokines and their receptors, and the capacity of glioma cells responding to cytokine stimulation, leading to inhibition of glioma cell proliferation. Conceivably, induction of LRRC4 expression may provide new intervention for human glioma in the clinic.
Wu
Minghua
M
Cancer Research Institute, Central South University, Changsha, Hunan, The People's Republic of China.
Huang
Chen
C
Li
Xiayu
X
Li
Xiaoling
X
Gan
Kai
K
Chen
Qiong
Q
Tang
Yunlian
Y
Tang
Ke
K
Shen
Shourong
S
Li
Guiyuan
G
D015153:Blotting, Western; D018929:Cell Culture Techniques; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002999:Clone Cells; D016207:Cytokines / Q000378:metabolism*; D015536:Down-Regulation; D042783:Endothelial Cells / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005562:Formazans / Q000378:metabolism; D005909:Glioblastoma / Q000235:genetics / Q000473:pathology*; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000494:pharmacology; D007668:Kidney / Q000166:cytology; D008156:Luciferases / Q000378:metabolism; D009389:Neovascularization, Pathologic / Q000188:drug therapy*; D009419:Nerve Tissue Proteins / Q000378:metabolism* / Q000494:pharmacology; D020411:Oligonucleotide Array Sequence Analysis; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D012334:RNA, Neoplasm / Q000302:isolation & purification; D013778:Tetrazolium Salts / Q000378:metabolism; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
D016207:Cytokines; D005562:Formazans; C503331:LRRC4 protein, human; D009419:Nerve Tissue Proteins; D010982:Platelet-Derived Growth Factor; D012334:RNA, Neoplasm; D013778:Tetrazolium Salts; D042461:Vascular Endothelial Growth Factor A; C079782:MTT formazan; D004815:Epidermal Growth Factor; D007335:Insulin-Like Growth Factor II; D008156:Luciferases