Correlation of DACT2 expression with clinicopathological parameters In this study, we also included another cohort of 61 HCC patients who had undergone liver transplan tation. We divided them into two expression groups. The low expression group included pa tients whose tumor tissue expressed DACT2 mRNA below the median selleckbio level. The high expression group Inhibitors,Modulators,Libraries consisted of patients whose tumor tissue expressed DACT2 mRNA above the median value. Clini copathological parameters are listed in Table 1 in relation to DACT2 mRNA expression status. In the relationship between Inhibitors,Modulators,Libraries DACT2 expression and the clinicopathological data, we found that DACT2 expression was negatively correlated with tumor size. Tumors with low DACT2 expression were significantly larger in size than those with high expression.

There was no statistically significant correlation between DACT2 expression and age, gender, AFP, histopathological Inhibitors,Modulators,Libraries gra ding, or tumor number. Downregulation and promoter methylation of DACT2 in liver cancer cells We first detected the expression level of DACT2 in five liver cancer cell lines. The results showed that DACT2 Inhibitors,Modulators,Libraries transcript was silenced or reduced in HepG2 and HCCLM3 cells. To examine the role of promoter methy lation in the silencing of DACT2, we further analyzed the methylation status of the DACT2 gene the results indicate that the DACT2 gene promoter was partially methylated in cell lines with reduced or silenced expression. To demonstrate whether methylation mediates DACT2 si lencing, we treated two methylated cell lines that showed silencing of DACT2 with 5 Aza.

5 Aza restored DACT2 expression in both cell lines with obvious demethylation observed in the treated HepG2 cell line by bisulfite genomic sequencing, suggesting that DNA methylation plays a crucial role Inhibitors,Modulators,Libraries in the transcriptional silencing of DACT2 in liver cancer cells. Cell cycle alterations after DACT2 silencing As shown in Figure 3A, the inhibitory efficiency of siRNAs for gene transcription was significant in morderate metastasic potential MHCC97L cells. To determine the role of DACT2 in tumor cell cycle progression and tumor cell proliferation, we examined the effect of DACT2 knock down in MHCC97L cells on the cell cycle. After 48 h of transfection, cell proliferation as determined by the number of cells in the S phase was increased from 25. 9% to 34. 6%, as is shown in Figure 3B,C.

Moreover, the num ber of cells in the else G0G1 phase decreased from 61. 7% to 50. 5%. DACT2 silencing promotes cell migration and invasion The effect of DACT2 on the migration and invasiveness of HCC cells was analyzed using the Matrigel model. We found that the average number of migratory and invaded cells transfected with DACT2 siRNA was significantly increased when compared to those with negative control siRNA, indicating that the invasive potential of MHCC97L cells increased after DACT2 knockdown.