Original Articles
Copyright ©The Author(s) 1998. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 15, 1998; 4(4): 294-297
Published online Aug 15, 1998. doi: 10.3748/wjg.v4.i4.294
Expression of alpha fetoprotein messenger RNA in BEL-7404 human hepatoma cells and effect of L-4-oxalysine on the expression
Xing-Wang Wang, Bin Xu
Xing-Wang Wang, Bin Xu, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 200031, China
Xing-Wang Wang, male, born on July 23, 1959 in Yangzhou, Jiangsu, China, graduated from Zhejiang Medical University in 1980. From 1987 to 1990, under the instruction of Professors Xu Shu-Yun and Chen Ming Zhu, he was engaged in immunopharmacological studies and obtained Master of Sciences. From 1994 to 1997, under the instruction of Professor XU Bin, he was engaged in the studies of cancer pharmacology and obtained M.D. and Ph.D. Now he is working in Shanghai Institute of Cell Biology, Chinese Academy of Sciences as postdoctor and associate professor. He has published more than 15 papers in the international journals and more than 40 papers in the Chinese journals as well as 8 books (one book as editor-in-chief).
Author contributions: All authors contributed equally to the work.
Correspondence to: Dr. Xing-Wang Wang, Shanghai Institute of Cell Biology, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai 200031, China
Received: March 8, 1998
Revised: May 12, 1998
Accepted: June 9, 1998
Published online: August 15, 1998
Abstract

AIM: To investigate alpha-fetoprotein (AFP) mRNA expression in BEL-7404 human hepatoma cells and the effect of L-4-oxalysine (OXL) on the expression.

METHODS: Bel-7404 human hepatoma cells were maintained in RPMI 1640 media. Human AFP cDNA probe was labelled with digoxigenin-11-dUTP by the random primer labelling method. The expression of AFP mRNA in Bel-7404 cells was determined by an in situ hybridization technique with digoxigenin-labelled human AFP cDNA probe. The positive intensities of AFP mRNA in cells were analyzed by microspectrophotometer and expressed as absorbance at 470 nm. For the experiment with OXL, cells were incubated with various concentrations of the agent for 72 h.

RESULTS: Essentially all the hepatoma cells contained AFP mRNA in the cytoplasm, although in various amounts. The specificity of the hybridization reaction was confirmed by control experiments in which the use of Rnase-treated BEL-7404 cells, non-AFP producing cells (HL-60 human leukemia cells) or a nonspecific cDNA probe resulted in negative hybridization. When the cells were treated with OXL (25, 50 mg/L), the content of AFP mRNA in the cytoplasm was decreased with the inhibition percentages of 34.3% and 70.1%, respectively (P < 0.05).

CONCLUSION: AFP mRNA was expressed in BEL-7404 human hepatoma cells and OXL suppressed AFP mRNA expression in the cells.

Keywords: oxalysine; liver neoplasms; fetoprotein; tumor cell, cultured; RNA, messenger; gene expression; in situ hybridization; immunohistochemistry