Knockdown of GCF2/LRRFIP1 by RNAi causes cell growth inhibition and increased apoptosis in human hepatoma HepG2 cells.

Guo-Rong Luo,Nai-Xia Cao,Xiao-Xun Xie,Ri-Ting Jiang,Tian-Ming Huang,Shao-Jian He,Li Chen,Bo Wu,Su-Fang Zhou,Jing-Ping Li,Ping Ma,De-Feng Chen

doi:10.7314/apjcp.2014.15.6.2753

Guo-Rong Luo, Nai-Xia Cao + Show 10 more

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https://doi.org/10.7314/apjcp.2014.15.6.2753

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Abstract

GC-binding factor 2 (GCF2) is a transcriptional regulator that represses transcriptional activity of the epidermal growth factor receptor (EGFR) by binding to a specific GC-rich sequence in the EGFR gene promoter. In addition to this function, GCF2 has also been identified as a tumor-associated antigen and regarded as a potentially valuable serum biomarker for early human hepatocellular carcinoma (HCC) diagnosis. GCF2 is high expressed in most HCC tissues and cell lines including HepG2. This study focused on the influence of GCF2 on cell proliferation and apoptosis in HepG2 cells. GCF2 expression at both mRNA and protein levels in HepG2 cells was detected with reverse transcription (RT) PCR and Western blotting, respectively. RNA interference (RNAi) technology was used to knock down GCF2 mRNA and protein expression. Afterwards, cell viability was analyzed with a Cell Counting Kit-8 (CCK-8), and cell apoptosis and caspase 3 activity by flow cytometry and with a Caspase 3 Activity Kit, respectively. Specific down-regulation of GCF2 expression caused cell growth inhibition, and increased apoptosis and caspase 3 activity in HepG2 cells. These primary results suggest that GCF2 may influence cell proliferation and apoptosis in HepG2 cells, and also provides a molecular basis for further investigation into the possible mechanism at proliferation and apoptosis in HCC.

Highlights

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors of the digestive system (Venook et al, 2010)
Results small interference RNA (siRNA)-1 was the most effective siRNA The reverse transcription (RT)-PCR analysis results showed the relative level of GC-binding factor 2 (GCF2) mRNA in HepG2 cells transfected with siRNA-1 was lowest in all HepG2 cells transfected with different siRNAs or controls at 48 hours after transfection (Figure 1A)
The results revealed that GCF2 protein expression was decreased at 48 hours after transfection with siRNA-1 (Figure 1B)

Summary

Introduction

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors of the digestive system (Venook et al, 2010). GCF2 was found as the key mediator for the Wnt pathway, the knockdown of GCF2 repressed remarkably cancer cell adhesion, migration and invasion (Ohtsuka et al, 2011; Ariake et al, 2012) These reports above suggest the functions of GCF2 may be multifarious and closely related to biological behaviors of tumors. GC-binding factor 2 (GCF2) is a transcriptional regulator that represses transcriptional activity of the epidermal growth factor receptor (EGFR) by binding to a specific GC-rich sequence in the EGFR gene promoter. In addition to this function, GCF2 has been identified as a tumor-associated antigen and regarded as a potentially valuable serum biomarker for early human hepatocellular carcinoma (HCC) diagnosis. Conclusions: These primary results suggest that GCF2 may influence cell proliferation and apoptosis in HepG2 cells, and provides a molecular basis for further investigation into the possible mechanism at proliferation and apoptosis in HCC

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Conclusion