Abstract
Objective: Transcription factor SIX1 aberrant expression has been shown in various mammalian tumors, and also recent studies indicated that SIX1 has a role in cancer stem cell properties. However, its roles in HCC cancer stem cell characteristics remain unclear. The aim of the study, to evaluate the EMT-inducer SIX1 and cancer stem cell markers expression profile in-vitro and in-vivo analyses.
 Material and Methods: SIX1 expression was suppressed by short hairpin RNA transduction in the SNU398 HCC cell line. Tumorsphere formation assay is a golden useful assay for cancer stem cell analysis. SIX1-dependent cancer stem cell markers PROM1, EPCAM, and OCT4 differential gene expression profiles were assessed in tumorsphere formation assay by RT-qPCR. Differential expression and correlation analyses were performed in transcriptome data in cirrhosis and HCC tissue samples.
 Results: Assessment of SIX1-knockdown expression of target genes in tumorsphere formation assay revealed PROM1, EPCAM expressions were significantly up-regulated but OCT4 gene expression was significantly down-regulated. Conformably, PROM1 and EPCAM expressions were inversely but OCT4 expression was positively correlated in transcriptome arrays in HCC tissues. Interestingly, to the evaluation of the same gene expressions were shown different patterns except, OCT4 in cirrhosis samples. The mRNA expression profiles did not change between cirrhosis and HCC samples in the SIX1, EPCAM, PROM1, and OCT4 gene expression profiles.
 Conclusion: Cancer stem cells are self‐renewable cell types and are responsible for cancer progression. Findings from this study highlight the SIX1 and cancer stemness-related genes expression correlations to improve our knowledge for HCC molecular signatures.
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