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Antisense cathepsin B RNA inhibits invasion and metastasis of human breast carcinoma cells
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Abstract:
Abstract Objective:To investigate the effect of antisense cathepsin B(CatB) RNA on the invasion and migration of human breast carcinoma cells. Methods: PBudCE4.1-antiCatB recombinant plasmid carrying antisense cathepsin B(CatB) gene was constructed and transfected into breast carcinoma cell line MDA-MB-231 by lipofection system. The expression of CatB protein in MDA-MB-231 cells was detected by Western blotting analysis; the proliferation of MDA-MB-231 cells was determined by MTT assay. Cell-matrix adhesion assay was used to examine the effect of anti-CatB on adhesion ability of MDA-MB-231 cells. The effects of anti-CatB on the invasion and migration abilities of MDA-MB-231 cells were measured by invasion and migration transwell system. Results: The recombinant plasmid PBudCE4.1-antiCatB was successfully constructed. Expression of CatB protein in MDA-MB-231 cells was decreased after PBudCE4.1-antiCatB transfection compared with those in untransfected and mock-vehicle transfected cells([0.96±0.02] vs [1.98±0.23], [1.84±0.08], P<0.05); the proliferation of MDA-MB-231 cells was also inhibited in PBudCE4.1-antiCatB transfected group([0.255±0.017] vs [0.458±0.033], [0.421±0.022], P<0.01); and the adhesion abilities(binding to matrix or fibronectin) were decreased([0.054±0.017] vs [0.111±0.018], [0.107±0.017], P<0.01; [0.052±0.008] vs [0.120±0.014], [0.113±0.009], P<0.01). Transwell asaay showed that the invasion and migration abilities were inhibited in PBudCE4.1-antiCatB transfected group compared with those in the non-transfection and mock-vehicle transfected groups([52.80±7.76] vs [124.00±44.54], [116.80±32.87], P<0.01; [60.25±8.73] vs [132.50±12.15], [119.20±25.13], P<0.01). Conclusion: The expression of antisense CatB RNA can inhibit the growth, adhesion, invasion and migration abilities of MDA-MB-231 cell in vitro.
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Project Supported:
Project supported by the Natural Science Foundation of Fujian Province(No. X0650058), and the Higher Institute Foundation of Fujian Province(No. 2006F5047)
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