Figure 5. Suppressed TTL expression in stromal cells of human colon and liver cancer tissues.

(A) Paraffin-embedded tissue samples ± 1 cm from the boundary of a tumor nodule (tumor area) or more than 2 cm away from tumor nodules (non-malignant areas) of human colon carcinoma were stained by immunohistochemistry for TTL (brown). (Upper left panel) Representative images from colon areas of one patient; boxed areas enlarged next to original (upper right panel); G, colonic glands; S, stroma; scale bars, 20 μm. (B) Representative images of TTL⁺ cells (brown) stained in non-malignant and adenoma areas of the colon tissues from a mouse model of intestinal inflammation-induced carcinogenesis (AOM/DSS). G, colonic glands; S, stromal surrounding the colonic glands; scale bars, 20 μm. (C) Tumor and non-tumor areas as described in (A) from human hepatocarcinoma tissues were stained for detyrosinated-tubulin (abbreviated as “Glu-tublin”, brown). Representative images from liver tissue areas from one patient. T, tumor nests; S, stroma. Bars, 50 μm. (A-C, lower panels) Proportions of TTL⁺ or detyrosinated-tubulin⁺ stromal cells were calculated by counting TTL⁺ or detyrosinated-tubulin⁺ stromal cells and all nuclei within the stroma from 35 visual fields (AOM/DSS induced), or 6 visual fields (human colon or liver) tissue sample. Mean ± SEM, ^**P< 0.01, Student’s t test, n=6 tumors per group. (D) Human TTL mRNA levels in non-malignant colon tissues (n = 72) and colon tumor tissues (n = 77). Scatter plot and mean, ^**P < 0.0001, Student’s t test. Cut-off value for definition of TTL^high samples set as 2.0. (E) TTL^low and TTL^high expression in colon tumor tissue was correlated to the overall survival of the colon tumor patients after surgery. P < 0.01, log-rank test, n = 72. (F) The TTL expression levels in colon tumor tissues from T3 (n = 42) and T4 stage (n = 16) were compared. Scatter plot and mean, ^*P= 0.012, Student’s t test. (G) TTL^low and TTL^high expression in liver tumor tissues was defined as described in (D) and correlated to the overall survival times of the hepatocarcinoma patients after surgery. P= 0.0243, log-rank test, n = 31.