Gastric Cancer
Copyright ©The Author(s) 2002. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 15, 2002; 8(2): 213-216
Published online Apr 15, 2002. doi: 10.3748/wjg.v8.i2.213
Applying a highly specific and reproducible cDNA RDA method to clone garlic up-regulated genes in human gastric cancer cells
Yong Li, You-Yong Lu
Yong Li, You-Yong Lu, Beijing Institute for Cancer Research, Beijing Laboratory of Molecular Oncology, School of Oncology, Peking University, Beijing 100034, China
Author contributions: All authors contributed equally to the work.
Supported by the Natural Scientific Foundation of China (NSFC3962526) and National High-Technology Project-863 (102-10-01-04)
Correspondence to: Dr. You-Yong Lu, Beijing Institute for Cancer Research, Beijing Laboratory of Molecular Oncology, School of Oncology, Peking University, 1 Da-Hong-Luo-Chang Street, Western District, Beijing 100034, China.yongylu@public.bta.net.cn
Telephone: +86-10-66163061 Fax: +86-10-66175832
Received: September 14, 2001
Revised: October 15, 2001
Accepted: October 21, 2001
Published online: April 15, 2002
Abstract

AIM: To develop and optimize cDNA representational difference analysis (cDNA RDA) method and to identify and clone garlic up-regulated genes in human gastric cancer (HGC) cells.

METHODS: We performed cDNA RDA method by using abundant double-stranded cDNA messages provided by two self-constructed cDNA libraries (Allitridi-treated and paternal HGC cell line BGC823 cells cDNA libraries respectively). BamH I and Xho I restriction sites harbored in the library vector were used to select representations. Northern and Slot blots analyses were employed to identify the obtained difference products.

RESULTS: Fragments released from the cDNA library vector after restriction endonuclease digestion acted as good marker indicating the appropriate digestion degree for library DNA. Two novel expressed sequence tags (ESTs) and a recombinant gene were obtained. Slot blots result showed a 8-fold increase of glia-derived nexin/protease nexin 1 (GDN/PN1) gene expression level and 4-fold increase of hepatitis B virus x-interacting protein (XIP) mRNA level in BGC823 cells after Allitridi treatment for 72 h.

CONCLUSION: Elevated levels of GDN/PN1 and XIP mRNAs induced by Allitridi provide valuable molecular evidence for elucidating the garlic's efficacies against neurodegenerative and inflammatory diseases. Isolation of a recombinant gene and two novel ESTs further show cDNA RDA based on cDNA libraries to be a powerful method with high specificity and reproducibility in cloning differentially expressed genes.

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