Fucosylation, which is catalyzed by fucosyltransferases (FUTs), is one of the most important glycosylation events involved in cancer. 0.05, ** 0.01, significant difference between groups as indicated. The protein expression of FUT2/FUT8 in lung cancer and lung adenocarcinoma To investigate the protein levels in lung cancer and lung adenocarcinoma tissues, Western Blot was used to examine the protein level of FUT2 and FUT8 (Figure ?(Figure2A).2A). Compared to adjacent noncancerous tissue, the FUT2 expression was significantly increased in lung cancer (n=21) (Figure ?(Figure2B)2B) and lung adenocarcinoma tissues (n=15) (Figure ?(Figure2C).2C). The FUT8 expression was also markedly increased in lung cancer (n=13) (Figure ?(Figure2D)2D) and lung adenocarcinoma specimens (n=9) (Figure ?(Figure1E),1E), compared with adjacent noncancerous tissue. These data indicated that both FUT2 and FUT8 were significantly up-regulated in cancer lung and lung adenocarcinoma. Open in a separate window Figure 2 The protein expression levels of FUT2 and FUT8 in lung cancer(A) Western Blot analysis of FUT2 and FUT8 levels in tumor tissues. (B) (D) The protein expression levels of FUT2/FUT8 in lung F2r cancer. (C) (E) The protein expression levels of FUT2/FUT8 in lung adenocarcinoma. Tubulin was acted Volasertib cost as a loading control. N: adjacent noncancerous tissues; C: cancer tissue. * 0.05, ** 0.01, significant difference between groups as indicated. The expression of FUT2 in lung adenocarcinoma To explore the expression of FUT2 in lung adenocarcinoma patients, we performed immunohistochemical (IHC) staining on slides with lung adenocarcinoma tissue, including stage I, II and III. As shown in Figure ?Figure3,3, tissue histology showed that cancer cells were pleomorphism, and most of them were the solid mass or small funicular line, even the lumens is forming, and arranged in a tubular adenoid structure (Figure ?(Figure3A).3A). FUT2 was only Volasertib cost slightly detectable in lung tissues by immunostaining, and increased expression of FUT2 was detected in all stages of lung adenocarcinoma tissues (Figure ?(Figure3B).3B). However, there was no distinct difference among the clinical Volasertib cost stages. Statistical analysis was performed to examine the correlation the FUT2 protein level represented by staining intensity, and showed in Figure ?Figure3C3C and ?and3D.3D. The expression of Volasertib cost FUT2 in lung adenocarcinoma tissues was higher than that in adjacent noncancerous tissue ( Volasertib cost 0.0001), but there were no difference among the clinical stages I, II and III ( 0.05). These data indicate that FUT2 is significantly increased in lung adenocarcinoma with no correlated with clinical stages. Open in a separate window Figure 3 The expression levels of FUT2 protein in lung adenocarcinoma(A) Tissue histology revealed tumors morphological characteristics: 200. (B) Representative images of FUT2 staining using IHC assay in normal adjacent tissue and 74 cases of archived lung adenocarcinoma specimens with different clinical stages: 200. (C) (D) Quantitative analysis of the average MOD of FUT2 staining in normal adjacent tissue and lung adenocarcinoma specimens. (E) The statistics of protein expression of FUT2 in cancer and normal tissue. N: Normal adjacent tissue; C: Cancer tissue; n: number of cases; CI: Cancer stage I, CII: Cancer stage II, CIII: Cancer stage III. **** 0.0001, significant difference between groups as indicated. The effect of FUT2 knockdown on cell migration and invasion of lung adenocarcinoma cells To further examine the effects of FUT2 on cell growth, migration and invasion, we generated stably clones suppressed FUT2 expression by vector-based transfection of the sh-FUT2 plasmid in A549 and H1299 cells. We also prepared control A549 and H1299 cells transfected using a scrambled vector (NC) to compare cell growth, migration and invasion by culture assays and xenograft model. Real-Time PCR showed that the mRNA expression of FUT2 was significantly reduced in A549 cells with sh-FUT2 (Figure ?(Figure4A).4A). Western Blot analysis validated that stable FUT2 RNAi clones significantly suppressed FUT2 in A549 and H1299 cells (Figure ?(Figure4B4B and ?and4C).4C). The expression level of FUT2 was quantified by densitometry using GAPDH as the loading control. Open in a separate window Figure 4 FUT2 knockdown inhibits migration and invasion of lung adenocarcinoma cells(A) FUT2 and GAPDH amplification curve and melting curve, and the relative FUT2 mRNA level. (B) The FUT2 protein.