Tumour invasion and metastasis are promoted in mice deficient in tenascin-X.

Abstract

Tenascin-X (TNX) is a member of the tenascin family of large oligomeric glycoproteins of the extracellular matrix (ECM). To determine whether TNX plays a part in tumour invasion and metastasis and to disclose its normal physiological role, we disrupted its gene in mouse embryonic stem cells by homologous recombination and created mice deficient in TNX. TNX-null mutant (TNX-/-) mice arose at normal frequency and showed no obvious defects during their adult life. However, when TNX-/- mice were subcutaneously inoculated in foot-pads with a highly invasive and metastatic cell line, B16-BL6 melanoma cells, the primary tumour size at 30 days after inoculation in the TNX-/- mice had increased by 1.2-fold compared with that in wild-type mice, and the invasion to the ankle and pulmonary metastasis in TNX-/- mice were also augmented by 2.2-fold and 6.8-fold, respectively, compared to those in wild-type mice. To disclose the molecular mechanism(s) of the promotion of tumour invasion and metastasis in TNX-/- mice, we measured the protein levels of matrix metalloproteinases (MMPs), which are recognized as playing a key role in these events, in the foot-pad homogenates of TNX-/- mice prior to the inoculation of melanoma cells. Gelatin zymography showed that the activities of proMMP-2, active MMP-2 and proMMP-9 were significantly higher in TNX-/- mice than in wild-type mice. Furthermore, a Northern blot analysis demonstrated that this increased activity of MMP-2 in TNX-/- mice was due to the induced expression of MMP-2 at the transcriptional level. The elevated expression of MMP-2 and MMP-9 resulted in decreased laminin levels, to less than half that of wild-type mice in the homogenates of TNX-/- mice. TNX deficiency led to an increase in the production of MMPs, and the increased activity of MMPs may result in the degradation of laminin. Consequently, the melanoma cells inoculated in TNX-/- mice might facilitate invasion and metastasis. These results imply that TNX is required for impeding the invasion and metastasis of tumour cells.