科研之友
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摘要
Transforming growth factor (TGF)-beta-inducible nuclear protein 1 (TINP1) functions to regulate cell proliferation. This study used bioinformatics techniques to analyze TINP1 cDNA and assessed the role of the nuclear localization signal 2 (NLS2) in the regulation of TINP1-induced cell proliferation. An NLS2-truncated TINP1 DNA fragment was generated and subcloned into the pcDB vector. Hela cells were transfected with pEGFP-N1-TINP1-Delta NLS2 and cellular localization of TINP1-Delta NLS2 was analyzed by fluorescence microscopy. HEK293T cells were co-transfected with luciferase reporter plasmids for proliferation-related genes. Expression of proliferation-related genes was also assessed by PCR and Western blot. The effect of TINP1-Delta NLS2 on proliferation was assessed by cell viability assay. TINP1-Delta NLS2 was found to not affect the nuclear localization of TINP1, but significantly enhanced expression of T-bet, NF-kappa B, AP-1, WNT, C/EBP and p53 (P<0.05) more strongly than TINP1. TINP1-Delta NLS2 transfection induced HeLa and HCT116 cell proliferation more weakly than TINP1. TINP1-Delta NLS2 transfection reduced levels of p53 and p21 mRNA and protein less strongly than TINP1, whereas TINP1-Delta NLS2 transfection substantially reduced expression of cyclin E mRNA and protein. In conclusion, TINP1-Delta NLS2 did not affect nuclear localization of TINP1, and possessed a weaker capacity to induce proliferation and modulate gene expression than TINP1, suggesting that NLS2 is required for TINP1 suppression of p53 signaling and TINP1-induced proliferation.
