Budwit\Novotny DA, McCarty KS, Cox EB, et al. HCC Upregulated KIF20B has been reported in some solid tumors, such as breast and bladder cancers.13 Here, we observed a significant upregulation of KIF20B in HCC tissues (n = 50) compared with adjacent (n = 36) or non\tumor (n = 14) tissues (Figure ?(Figure1A,B).1A,B). We further analyzed the data of 336 HCC CGP77675 and 42 non\tumor patients available in The Cancer Genome Atlas (TCGA) database to investigate whether expression correlates with HCC prognosis. Consistent with the IHC results, the mRNA levels of were much higher in HCC tissues compared with normal tissues (Figure ?(Figure1C).1C). Importantly, HCC patients with higher than median expression showed significantly shorter overall (= CGP77675 .036, left panel, Figure ?Figure1D)1D) and disease\free survival duration (= .022, right panel, Figure ?Figure1D),1D), especially in the early period, which Itga10 has a higher confidence level compared to the late stage. Together, these results suggest upregulated KIF20B in HCC tissues, and its expression level CGP77675 is negatively correlated with the prognosis of patients. Open in a separate window Figure 1 Overexpression of kinesin family member 20B (KIF20B) in hepatocellular carcinoma (HCC) samples. A, Representative pictures of KIF20B immunohistochemical staining on clinical samples. B, H\score of KIF20B for different groups; presented as mean + SD. C, mRNA levels of KIF20B in HCC and para\HCC tissues (data from The Cancer Genome Atlas [TCGA]). D, Overall (left panel) and disease\free (right panel) survival rates of HCC patients with high CGP77675 KIF20B expression levels (red) and low levels (blue) (the cut\off for determining high vs low levels of KIF20B is the midpoint, data from TCGA). (*< .05, ***< .001) 3.2. Reducing KIF20B sensitizes HCC cells to taxol CGP77675 Evidence has suggested that some KIF proteins are correlated with taxol resistance of cancer cells.25 To fully address whether reducing KIF20B increases the taxol sensitivity of HCC cells, we applied Ad\shKIF20B, a recombinant adenoviral vector expressing shRNAs against in HepG2, Hep3B and HuH\7 cell lines. Significantly enhanced taxol cytotoxicity was observed in all three cell lines receiving Ad\shKIF20B (Figure ?(Figure2B,C).2B,C). Furthermore, soft agar colony formation assay indicated that HCC cells receiving Ad\shKIF20B/taxol combined treatment showed markedly reduced colony numbers compared with the respective shKIF20B or taxol mono\treated cells (Figure ?(Figure2D).2D). Moreover, isobologram analysis suggested that the shKIF20B/taxol combination brings synergistic effects on suppressing the viability of these cell lines (Figure ?(Figure22E). Open in a separate window Figure 2 Adenoviral vector expressing small hairpin RNAs targeting kinesin family member 20B (Ad\shKIF20B) enhances taxol toxicity to hepatocellular carcinoma cells. A, Quantification of KIF20B mRNA levels in HepG2, Hep3B and HuH\7 cells 48 h after infection. MOI = 1. B, Relative cell viability of HepG2, Hep3B and HuH\7 cells by MTT assays 72 h after indicated treatments. C, Relative cell viability of HepG2, Hep3B and HuH\7 cells with indicated treatments by MTT assays. MOI = 1, taxol concentration = 1 mol/L. B,C, Value of control group was arbitrarily set at 1. Three independent experiments were carried out. D, Colony formation assays with indicated treatments. MOI = 1, taxol concentration = 1 mol/L. E, MTT assays were carried out after cells received adenoviral vector expressing shRNAs targeting KIF20B (Ad\shKIF20B) and taxol for 72 h. Standard isobolograms are shown. IC 50 values.