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Isolation of diallyl trisulfide inducible differentially expressed genes in human gastric cancer cells by modified
1Peking University, School of Oncology, Beijing, China.
Abstract:
Extensive epidemiologic studies indicated protective effects of consumption of garlic on reducing human gastric cancer (HGC) incidence. Diallyl trisulfide (DATS), a critical organic allyl sulfur component of garlic, was reported to have chemopreventive effects in inhibiting tumor process. We used DATS to treat HGC cell line BGC823 cells, and showed that DATS induces G1/S arrest and apoptosis in BGC823 cells demonstrated by a flow cytometric analysis. To further isolate DATS inducible differentially expressed genes in BGC823 cells, we combined a highly specific subtractive hybridization of cDNA representational difference analysis (cDNA RDA) with a sensitive bidirectional radioactive detection of mRNA differential display (mRNA DD) to develop a subtractive hybridization differential display (SHDD) method. This modified method adopted a first round of bidirectional subtractive hybridization between two sample cDNAs and a second round of bidirectional subtractive hybridization between the two resultant first-round difference products. Bidirectional subtractive hybridizations magnified the differences between the two sample cDNAs and favored isolating mRNA species with very small expression differences. We employed the SHDD method to detect DATS inducible differentially expressed genes in BGC823 cells. A total of 14 cDNA fragments (11 upregulated and 3 downregulated by DATS treatment) were isolated and confirmed by reverse Northern blot analysis. Our data show that SHDD is a powerful technique for identifying differentially expressed mRNA species between two sample cDNAs and provide useful cellular and molecular information for understanding the effects of garlic against human gastric cancer.
Insights
Garlic compound diallyl trisulfide (DATS) shows promise in fighting human gastric cancer (HGC) by inducing cell cycle arrest and apoptosis. A novel method, subtractive hybridization differential display (SHDD), identified key genes involved in this anti-cancer effect.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Epidemiologic studies suggest garlic consumption reduces human gastric cancer (HGC) risk.
- Diallyl trisulfide (DATS), a key garlic component, exhibits chemopreventive properties against tumor development.
Purpose of the Study:
- To investigate the effects of DATS on HGC cell line BGC823.
- To identify DATS-inducible differentially expressed genes in BGC823 cells using a novel method.
Main Methods:
- Treatment of BGC823 cells with DATS.
- Flow cytometry analysis to assess cell cycle arrest and apoptosis.
- Development and application of subtractive hybridization differential display (SHDD) for gene expression analysis.
Main Results:
- DATS treatment induced G1/S arrest and apoptosis in BGC823 cells.
- The SHDD method successfully identified 14 differentially expressed cDNA fragments (11 upregulated, 3 downregulated) in response to DATS.
- Reverse Northern blot analysis confirmed the isolated gene fragments.
Conclusions:
- DATS exhibits anti-cancer effects on human gastric cancer cells.
- SHDD is an effective technique for identifying differentially expressed genes, offering insights into garlic's chemopreventive mechanisms against HGC.
