Correlation between the overexpression of urokinase receptor isoform uPAR (D1D2) and hepatic cell malignant transformation

YINGQUN ZHOU,XIAOPING LǙ,LINGLING ZHAN,SHAN LI

doi:10.3892/mmr.2014.2006

Abstract

In order to provide an abundant source of specimen and to reveal the correlation between the overexpression of urokinase plasminogen activator receptor isoform uPAR (D1D2) and hepatic cell malignant transformation, the optimal liver cell culture method was selected from three cell culture methods to culture and separate out liver cells with a high density, high purity and high activity. The specimens were used to culture and assess the uPAR (D1D2) mRNA level in normal liver cells, liver cancer cells and para-carcinoma cells. In the present study, the correlation between the overexpression of uPAR (D1D2) and hepatic cell malignant transformation was discussed. When comparing the tissue block adherent method, liver cell grinding method and pancreatic enzyme digestion method, the liver tissue adherent method was found to be economical, simple and overall the optimal method for liver cell culture. This was used as a reference standard for cell culture. RT-PCR was used to determine isoform uPAR (D1D2) mRNA level in normal liver cells, para-carcinoma cells and liver cancer cells. The comparison of uPAR (D1D2) mRNA levels in normal liver cells, para-carcinoma cells and liver cancer cells, demonstrated that the brightness of the cells clearly increased in normal liver cells, para-carcinoma cells and liver cancer cells. The comparison of the cell grey values of three groups demonstrated a statistically significant difference (P<0.05). The liver tissue adherent method was able to produce liver cells with a high density, high purity and high activity, providing a sufficient source of specimen for our subsequent experiments. The electrophoresis results showed that: uPAR (D1D2) mRNA expression increased from normal liver cells to para-carcinoma cells to liver cancer cells, inferring that uPAR (D1D2) mRNA overexpression may be the result of changes in the conformation of the uPAR isoform. In addition, it is closely associated with abnormal cell signal transduction, which leads to clonal proliferation and abnormal differentiation of liver cells with malignant transformation in liver cells.

Highlights

Hepatocellular carcinoma (HCC) has one of the highest incidence rates of malignant tumors, ranking fifth in the world, with one-third occurring in China, where each year more than a half of newly suffering HCC patients are Chinese.The occurrence of HCC is associated with numerous factors
There were a small amount of cells, which adhered to the wall of the tube the day and the majority of the adherent cells were epithelial cells
There was no association between the expression of urokinase plasminogen activator receptor (uPAR) (D1D2) in HCC, Due to ethical issues, numerous human trials are not able to be performed

Summary

Introduction

Hepatocellular carcinoma (HCC) has one of the highest incidence rates of malignant tumors, ranking fifth in the world, with one-third occurring in China, where each year more than a half of newly suffering HCC patients are Chinese.The occurrence of HCC is associated with numerous factors. It has been verified that uPAR affects cell proliferation, migration and adhesion, and its expression is associated with the malignant degree of a tumor [1]. The interaction of uPAR with integrin α5β1, a fibronectin receptor, is able to activate FAK and ERK. The uPAR signal conduction pathway mediated by integrin α5β1 can promote tumor cell proliferation and participate in the regulation of cell proliferation and differentiation [2]. The analysis of biomolecular interactions demonstrates that the ZHOU et al: OVEREXPRESSION OF uPAR AND HEPATIC CELL MALIGNANT TRANSFORMATION lack of affinity is due to the increase in the dissociation rate of the D1D2 ligand complex [5]. The combination of uPAR with extracellular proteins and its interaction with certain membrane receptors, including the integrin and epidermal growth factor receptor, is a new focus of study [6]

Methods

Results

Conclusion