Tuftelin 1 (TUFT1), a protein functioning distinctively in different tissues, is reported to be elevated in several types of cancers and the elevation of TUFT1 is correlated with unfavorable clinicopathologic characteristics and poor survival. However, the involvement of TUFT1 in renal cell carcinoma (RCC) remains unknown. In the current study, we investigated the role of TUFT1 in RCC and potential underlying mechanisms. RT-PCR and Western blot analysis showed that both the mRNA and protein levels of TUFT1 were increased in primary RCC tissue and RCC cell lines. TUFT1 overexpression in RCC cells resulted in enhanced cell proliferation and migration while knockdown of TUFT1 by contrast decreased the growth and migration of the RCC cells, indicating TUFT1 expression is involved in RCC cell growth and migration. The involvement of TUFT1 in the epithelial-mesenchymal transition (EMT) of RCC cells was also determined by measuring the expression of EMT-related markers. Our data showed that TUFT1 overexpression promoted RCC cell EMT progression while knockdown of TUFT1 suppressed such process. Further signaling pathway inhibition assay revealed that TUFT1-induced RCC cell growth, migration and EMT was significantly suppressed by PI3K inhibitor, but not JNK or MEK inhibitors. In addition, TUFT1 overexpression enhanced the AKT phosphorylation, a key member of the PI3K signaling pathway, while PI3K inhibitor suppressed such process. Taken together, our study showed that TUFT1 expression was elevated in RCC and such elevation promoted the proliferation, migration and EMT of RCC cells in vitro, through PI3K/AKT signaling pathway. The findings of our current study imply that TUFT1 is involved in RCC tumorigenesis, and it may serve as a biomarker for RCC diagnosis and a potential target for RCC treatment.

To observe the inhibitory effect of plumbagin on the invasion and migration of glioma cells U-118 and to explore the possible mechanism of its action. U-118 cells were cultured in medium with different concentrations of plumbagin and the cells were counted and morphological changes were observed under the microscope after 24 h of incubation and the cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, the effect of plumbagin on cell migration was detected by cell scratching assay and the effect of cell invasion was detected by Transwell method. Western blotting was used to detect the expression of matrix metalloproteinase-2, matrix metalloproteinase-9 and p21 protein.The results showed that the morphology of U-118 cells could be changed by plumbagin and it was dependent on the concentration; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide results showed that plumbagin significantly inhibited the growth of U-118 cells and its inhibitory ability was positively correlated with the concentration; scratch assay results showed that the planar motility of cells decreased significantly with the increase of plumbagin concentration; Transwell Western blotting results showed that plumbagin administration could inhibit the expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 proteins in U-118 cells, but significantly increased the expression of P21 and there was a positive correlation with the concentration. Plumbagin can inhibit the proliferation of U-118 cells and suppress cell invasion and the mechanism may be related to the inhibition of matrix metalloproteinase-2, matrix metalloproteinase-9 and p21 expressio.