Interaction of testisin with maspin and its impact on invasion and cell death resistance of cervical cancer cells

Abstract

Previous studies have shown that testisin promotes malignant transformation in cancer cells. To define the mechanism of testisin-induced carcinogenesis, we performed yeast two-hybrid analysis and identified maspin, a tumor suppressor protein, as a testisin-interacting molecule. The direct interaction and cytoplasmic co-localization of testisin with maspin was confirmed by immunoprecipitation and confocal analysis, respectively. In cervical cancer cells, maspin modulated cell death and invasion; however, these effects were inhibited by testisin in parallel experiments. Of interest, the doxorubicin resistance was dramatically reduced by testisin knockdown ( P = 0.016). Moreover, testisin was found to be over-expressed in cervical cancer samples as compared to matched normal cervical tissues. Thus, we postulate that testisin may promote carcinogenesis by inhibiting tumor suppressor activity of maspin. Structured summary MINT- 7712215, MINT- 7712176: Testisin (uniprotkb: Q9Y6M0) binds (MI: 0407) to Maspin (uniprotkb: P36952) by pull down (MI: 0096) MINT- 7712188: Testisin (uniprotkb: Q9Y6M0) and Maspin (uniprotkb: P36952) colocalize (MI: 0403) by fluorescence microscopy (MI: 0416) MINT- 7712115: Testisin (uniprotkb: Q9Y6M0) physically interacts (MI: 0915) with Maspin (uniprotkb: P36952) by two-hybrid (MI: 0018) MINT- 7712162, MINT- 7712128: Maspin (uniprotkb: P36952) physically interacts (MI: 0915) with Testisin (uniprotkb: Q9Y6M0) by anti bait co-immunoprecipitation (MI: 0006) MINT- 7712147: Testisin (uniprotkb: Q9Y6M0) physically interacts (MI: 0915) with Maspin (uniprotkb: P36952) by anti tag co-immunoprecipitation (MI: 0007)