Microarray and Quantitative PCR Analysis of Gene Expression Profiles in Response to Treatment with Tomato Leaf Extract in MCF-7 Breast Cancer Cells

Abstract

We previously found cytotoxic effects of tomato leaf extract (TLE) on the MCF-7 breast cancer cell line. The aim of this study was to ascertain the molecular mechanisms associated with the usage of TLE as an anticancer agent by microarray analysis using mRNA from MCF-7 breast cancer cells after treatment with TLE for 1 hr and 48 hrs. Approximately 991 genes out of the 30,000 genes in the human genome were significantly (p<0.05) changed after the treatment. Within this gene set, 88 were significantly changed between the TLE treated cells and the untreated MCF-7 cells (control cells) with a cut-off fold change >2.00. In order to focus on genes that were involved in cancer cell growth, only twenty-nine genes were selected, either down-regulated or up-regulated after treatment with TLE. Microarray assay results were confirmed by analyzing 10 of the most up and down regulated genes related to cancer cells progression using real-time PCR. Treatment with TLE induced significant up-regulation in the expression of the CRYAB, PIM1, BTG1, CYR61, HIF1-α and CEBP-β genes after 1 hr and 48 hrs, whereas the TXNIP and THBS1 genes were up-regulated after 1 hr of treatment but down-regulated after 48 hrs. In addition both the HMG1L1 and HIST2H3D genes were down-regulated after 1 hr and 48 hrs of treatment. These results demonstrate the potent activity of TLE as an anticancer agent.

An Integrating Approach for Genome-Wide Screening of MicroRNA Polymorphisms Mediated Drug Response Alterations

“…We supposed that drug treatment may cause variation in the drug target genes' 3′ UTR to change the stability of the structure of miRNA/mRNA interaction. Some research showed that BTG1 was upregulated in response to treatment with tomato leaf extract (TLE) [23], recombinant bromelain [24], and insulin-like growth factor I (IGF-I) [25] in the MCF-7 breast cancer cell line, which contained a SNP (rs764927448) in BTG1 by query of the Cancer Cell Line Encyclopedia (CCLE). Moreover, we found that BTG1 loss is regulated by has-miR-1255b-5p through the wild-type SNP into variant-type SNP in Mir2Drug database (Table 1).…”

Analysis of Key Genes and Pathways Associated with Colorectal Cancer with Microarray Technology

“…Of course circulating proteins exhibit more usefulness in clinical applications and some have been validated, like cytokeratin 18 (Greystoke et al, 2012) and M2-pyruvate kinase (Meng et al, 2012 Takahashi et al report that MiR-148a can act as a predictive biomarker in patients with advanced colorectal cancer (Takahashi et al, 2012). Various technologies are utilized to carry out the researches, among which proteomic tool (de Wit et al, 2012) and microarray technology (Amid et al, 2012) are two powerful methods enabling global identification of potential biomarkers.…”