Abstract

BackgroundOur previous cDNA array results indicated KIAA0101 as one of the differentially expressed genes in human hepatocellular carcinoma (HCC) as compared with non-cancerous liver. However, it is necessary to study its expression at protein level in HCC and its biological function for HCC cell growth.MethodWestern blot and tissue array were performed to compare KIAA0101 protein expression level in paired human HCC and non-cancerous liver tissues from the same patients. Investigation of its subcellular localization was done by using dual fluorescence image examination and enriched mitochondrial protein Western blot analysis. The in vitro cell growth curve was used for examing the effect of over-expression of KIAA0101 in HCC cells. FACS was used to analyze the cell cycle pattern in KIAA0101 expression positive (+) and negative (-) cell populations isolated by the pMACSKKII system after KIAA0101 cDNA transfection.ResultsWestern blot showed KIAA0101 protein expression was down-regulated in HCC tissues as compared with their counterpart non-cancerous liver tissues in 25 out of 30 cases. Tissue array also demonstrated the same pattern in 161 paired samples. KIAA0101 was predominantly localized in mitochondria and partially in nuclei. KIAA0101 cDNA transfection could inhibit the HCC cell growth in vitro. In cell cycle analysis, it could arrest cells at the G1 to S phase transition.ConclusionKIAA0101 protein expression was down-regulated in HCC. This gene could inhibit the HCC cell growth in vitro and presumably by its blocking effect on cell cycle.

Highlights

  • Our previous cDNA array results indicated KIAA0101 as one of the differentially expressed genes in human hepatocellular carcinoma (HCC) as compared with non-cancerous liver

  • Western blot showed KIAA0101 protein expression was down-regulated in HCC tissues as compared with their counterpart non-cancerous liver tissues in 25 out of 30 cases

  • Since the alteration of KIAA0101 expression reported so far was based on mRNA transcription, we studied the KIAA0101 protein expression level in human HCC as compared with the matched non-cancerous liver tissues by using an antibody prepared in our laboratory, and further investigated its subcellular localization in HCC cells and its biological effect on HCC growth and cell cycle

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Summary

Introduction

Our previous cDNA array results indicated KIAA0101 as one of the differentially expressed genes in human hepatocellular carcinoma (HCC) as compared with non-cancerous liver. The development of HCC is a multi-factor in etiology, multi-step and multi-gene involvement in carcinogenesis and progression. A broad spectrum of genes have been involved in HCC development related to their genetic or epigenetic alteration, including p53[1], p16, p21[2], p27[3], betacatenin[4], PTEN[5] and Rb etc. Recent studies on functional genomics of HCC have further revealed that a number of genes with novel sequences and unclarified functions were involved in HCC development or progression [6]. Based on cDNA array, we found KIAA0101, designated as OEACT-1[7], as one of the genes with differential expression in HCC. Several reports described that alteration of KIAA0101 expression (page number not for citation purposes)

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