PMC

(A) TUFT1 mRNA expression in HEK293-mutTUFT1 cells was approximately 47% of the expression observed in HEK293-wtTUFT1 cells (*** P ≤ 0.001). ACTB and B2M were used as reference genes. (B) The variant suppressed also tuftelin protein expression, as the relative intensity of tuftelin in HEK293-mutTUFT1 cells was 51% of the relative intensity of tuftelin observed in HEK293-wtTUFT1 cells. The relative intensities were calculated from the ratio of tuftelin to beta-tubulin absolute intensities. The faint band in the control cells corresponds to the endogenous tuftelin recognized by the antibody.

Relationship between tuftelin 1 expression and prognosis of hepatocellular carcinoma. A: Kaplan-Meier analysis was performed to compare the overall survival (OS) between hepatocellular carcinoma (HCC) with high (n = 66) and low (n = 66) TUFT1; A1: The high (n = 19) and low (n = 40) tuftelin 1 (TUFT1) with OS at early HCC; A2: The high (n = 47) and low (n = 26) TUFT1 with OS at advanced HCC; B: Kaplan-Meier analysis was performed to compare the disease-free survival (DFS) between HCC with high (n = 66) and low (n = 66) TUFT1; B1: The high (n = 19) and low (n = 40) TUFT1 with DFS at early HCC; B2: The high (n = 47) and low (n = 26) TUFT1 with DFS at advanced HCC.

Effects of tuftelin 1 on healing, invasion, and migration of hepatocellular carcinoma cells. A: The scratch test of the MHCC-97H cells, the cell healing ability was decreasing after the high tuftelin 1 (TUFT1) cells were transfected with the TUFT1-shRNA3 plasmid, and A1, the relative cell healing ability was significantly inhibited compared with the control or negative sh-RNA for tuftelin 1 mRNA (sh-NC) cells (^bP < 0.001); B: The scratch test of the Hep3B cells, the cell healing ability was increased after the cells were transfected with the over-expressing TUFT1 plasmid; B1, the relative cell healing ability was significantly improved compared with the control or blank control (NC) cells (^bP < 0.001); C: The migration and invasion abilities of the MHCC-97H cells were analyzed by the transwell test; C1: The histograms of the cell migration and invasion abilities among the different groups (^bP < 0.001): D: The migration and invasion abilities of the Hep3B cells were analyzed by the transwell test; D1: The histograms of the cell migration and invasion abilities among the different groups (^bP < 0.001). Data are presented as mean ± standard error of the mean of at least three independent experiments. sh-RNA: sh-RNA3 for tuftelin 1 mRNA.

Tuftelin 1 expression with clinical staging of hepatocellular carcinoma. A, A1: Hepatocellular carcinoma (HCC) tissue with strongly positive staining; B, B1: Adjacent non-HCC tissue with negative staining, original magnifications of × 40 (scale bar: 50 μm) in A and B, and × 200 (scale bar: 50 μm) in A1 and B1; C, D: Tuftelin 1 (TUFT1) expression in different HCC staging; C1, D1: Low TUFT1 expression in the non-HCC tissues; C2-C5: The brown staining of TUFT1 gradually increases in the HCC tissues from stage I to IV (original magnification × 40); D2-D5: The staining of TUFT1 in the non-HCC tissues from stage I to IV (original magnification × 400). TNM: Tumor-node-metastasis.