Expression of antisense RNA to S100A4 gene encoding an S100-related calcium-binding protein suppresses metastatic potential of high-metastatic Lewis lung carcinoma cells

Abstract

S100A4 (also known as pEL98/mts1/p9Ka/18A2/42A/ calvasculin/FSP1/CAPL), a member of S100-related calcium-binding proteins, has been implicated to play a role in metastasis. In the present study, we examined the e ect of antisense S100A4 RNA on metastatic potential of Lewis lung carcinoma (LLC) cells. High-metastatic A11 cells were transfected with the expression vector containing S100A4 cDNA in an inverted (antisense) orientation under the transcriptional control of the mouse metallothionein promoter. Treatment of a stably transfected clone (AS10 cells) with Zn 2+ resulted in the suppression of the experimental metastatic ability, which was accompanied with the expression of antisense S100A4 RNA and the suppression of the S100A4 expression at both the mRNA and the protein levels. To further con®rm the e ect of antisense S100A4 RNA, we established several clones after retroviral transduction with an antisense S100A4 construct. Notably, reduced metastatic potential was also evident in these clones. In the antisense S100A4 RNA-expressing cells, cell motility and in vitro invasiveness were found to be suppressed.

Suppression of metastasis-associated S100A4 gene expression by gamma-interferon in human colon adenocarcinoma cells

“…Conversely, the expression of antisense S100A4 mRNA in S100A4-expressing Lewis lung carcinoma cells lowered cell motility (Takenaga et al, 1997a). These results collectively support the assumption that S100A4 participates in regulating cell motility.…”

“…Cell motility was assessed by phagokinetic track assay as described elsewhere (Takenaga et al, 1994a(Takenaga et al, , 1997a. Cells were incubated for 24 h in growth medium containing 300 IU ml -1 IFN-γ, washed, and then seeded onto coverslips coated with 10 µg ml -1 BSA and colloidal gold particle.…”

“…We and others have shown that the increased expression of S100A4 in tumour cells by introduction of the S100A4 gene resulted in an enhancement of cell motility and invasive and metastatic potential of tumour cells (Davies et al, 1993;Parker et al, 1994;Takenaga et al, 1994a). On the other hand, reduction of the expression of S100A4 by means of antisense S100A4 RNA or anti-S100A4 ribozyme in high-metastatic tumour cells resulted in the suppression of their invasive and metastatic capabilities (Grigorian et al, 1993;MaeLandsmo et al, 1996;Takenaga et al, 1997a). Although the precise functions of the S100A4 protein are not fully understood, recent studies implicate that the protein interacts with cytoskeletal proteins such as non-muscle tropomyosins (Takenaga et al, 1994c(Takenaga et al, , 1994d and myosin (Kriajevska et al, 1994; in a Ca 2+ -dependent manner and thereby modulates cell motility (Takenaga et al, 1994a(Takenaga et al, , 1994d.…”

Network-based analysis of calcium-binding protein genes identifies Grp94 as a target in human oral carcinogenesis

“…A number of Ca 2 þ -binding proteins are thought to be implicated in establishing the malignant and metastatic phenotypes of various tumours (Chen et al, 1997;Takenaga et al, 1997;Van Ginkel et al, 1998), and recent studies have reported altered expression of several Ca 2 þ -binding protein genes in a wide range of human malignancies (Bustin et al, 2001;Pietas et al, 2002;Lakshmikuttyamma et al, 2004;Imazawa et al, 2005). Regarding oral squamous cell carcinomas (OSCCs), aberrant expression of S100 Ca 2 þ -binding protein families seen in tumour sites has been correlated with cancer cell invasion or metastasis (Moriyama-Kita et al, 2004;Tsai et al, 2005).…”

Extracellular S100A4(mts1) stimulates invasive growth of mouse endothelial cells and modulates MMP-13 matrix metalloproteinase activity

“…The S100A4 gene was originally isolated as a gene differentially expressed in highly metastatic mouse mammary adenocarcinoma cells (Ebralidze et al, 1989). Introduction of the S100A4 gene into nonmetastatic tumor cell lines and suppression of its activity in metastatic ones proved its involvement in metastasis formation Maelandsmo et al, 1996;Takenaga et al, 1997;Lloyd et al, 1998;Uozumi et al, 2000). Stimulation of tumor metastasis was demonstrated in two transgenic mouse models with overexpression of the S100A4 gene Davies et al, 1996).…”