no. silencing of disrupts mitochondrial function of melanoma cells, conferring a new target for melanoma removal. A recent work also revealed that is required for melanoma cell viability [26]. However, the role of in liver malignancy and liver TICs is usually unclear. Here we found high expression of in liver malignancy and liver TICs. initiates Wnt/-catenin activation and the self-renewal of liver TICs. interacts with and recruits EZH2 to the promoter of CTNNBIP1, and inhibits its transcription initiation. RESULTS is highly expressed in liver cancer and liver TICs plays an essential role in melanoma cell viability and metabolic vulnerability, while, its role in liver tumorigenesis and liver TIC self-renewal is usually unknown. Accordingly, we detected the expression levels of in liver cancer. is usually highly expressed in HCC samples, especially in advanced HCC samples (Physique 1A, 1B). Vitamin K1 Interestingly, only a small subset cells showed high expression of in HCC samples, especially in early HCC samples (Physique ?(Figure1B1B). Open in a separate window Physique 1 is highly expressed in live malignancy and liver TICs(A) RNA were extracted from 19 peri-tumor samples, 7 early hepatocellular carcinoma (HCC) and 12 advanced HCC (aHCC) samples, followed by realtime PCR detection for expression. The average expression level of peri-tumor samples was defined as 1. (B) expression profiles in peri-tumor, early HCC (eHCC) and advanced HCC (aHCC). Left panels were common images of hybridization, right panels were Vitamin K1 statistical results. Level bars, 50 m. (C) Liver TICs were enriched from main cells using TIC surface marker CD133, followed by detection with realtime PCR. Vitamin K1 (D) Liver oncospheres and non-spheres were collected for detection with realtime PCR. expression levels in non-sphere served as control. (E) Fluorescence hybridization (FISH) results showed high expression of in oncospheres. c-Myc served as a positive control. Level bars, 10 m. (F) Nuclear-cytoplasmic segregation was performed and subcellular location of was detected with realtime PCR (left panels). The efficiency of nuclear-cytoplasmic segregation was detected with Western blot (right panels). Data were shown as meanss.d. **P 0.01; ***P 0.001 by two-tailed Students t test. Data are representative of three independent experiments. We then enriched liver TICs from HCC primary samples using TIC surface marker CD133, followed by mRNA detection. Compared with CD133- non-TICs, TICs showed high expression (Figure ?(Figure1C).1C). Similarly, high expression of was also found in oncospheres (Figure ?(Figure1D).1D). We then confirmed expression profiles in oncospheres and non-spheres with fluorescence hybridization (FISH), and found was highly expressed in spheres (Figure ?(Figure1E).1E). To our surprise, was mainly located in nucleus according to Vitamin K1 ISH and FISH data (Figure 1B, 1E), but not mitochondrial localization. Accordingly, we performed nuclear-cytoplasmic segregation and examined the subcellular location of was highly expressed in liver cancer and liver TICs, with nuclear location. is required for liver TIC self-renewal We then explored the KLK3 role of in TIC self-renewal. Sphere formation assay is the most widely-accepted experimental system to detect TIC self-renewal. So we knockdown with antisense oligos (ASO), followed by sphere formation. silenced cells showed impaired sphere formation (Figure ?(Figure2A).2A). We further examined the long-term self-renewal of silenced TICs, and also found attenuate self-renewal in long-term incubation (Figure ?(Figure2B).2B). These data indicate the critical role of in liver TIC self-renewal. Open in a separate window Figure 2 is required for liver TIC self-renewal(A) silenced cells were established with antisense oligos (ASO) (left panels), followed by sphere formation assay. Typical photos of oncospheres were shown in middle panels and sphere initiating ratios were shown in right panels. (B) Sequential sphere formation assay were performed with silenced cells. 1st, the first passage; 2nd, the second passage; 3rd, the third passage; 4th, the fourth passage. (C) 1104 silenced and control cells were used for transwell assay, and invasive cells were visualized Vitamin K1 by crystal.