Down-regulated expression of cytochrome P450 related genes involved in hepatocellular carcinoma

Abstract

Objective: To analyze the differentially expressed genes (DEGs) among hepatocellular carcinoma (HCC), para-cancerous tissue (PCT) and normal liver tissue (NLT) by using Agilent microarray and explore the target genes related to the development and progression of HCC. Methods: Total RNA of matched HCC, PCT and NLT of HCC patients was isolated using one step Trizol method and qualified using Lab-on-chip method. cRNAs were synthesized, fluoresce labeled and purified after total RNAs purified. The RNAs of HCC and NLT, HCC and PCT were hybridized to Agilent microarray (21 074 probes) respectively. The fluorescence intensities were detected by Agilent scanner and quantified by Feature Extraction software. The DEGs which both differentially expressed in HCC vs NLT and HCC vs PCT were selected as candidate genes and confirmed by SYBR Green I stained real time RT-PCR. Results: (1) The total RNAs, reverse transcription products and fluoresce labeled cRNAs were of high quality; (2) There were 420 up-regulated genes and 552 down-regulated genes in 2-fold DEGs, including 23 genes related to cytochrome P450; (3) CYP2C8 was selected as a candidate gene of 10-fold down-regulated genes. The results of real time RT-PCR, using β-actin as an internal control, revealed that the 2-ΔCt values of CYP2C8 in HCC, PCT and NLT were 0.009383, 0.316812 and 0.607182, respectively. Conclusion: (1) The high throughput and effective Agilent oligomicroarray can screen novel therapy target genes by analyzing the DGEs in development and progression of HCC; (2) The development and progression of HCC is a complicated process involving multigenes and multiprocedures; (3) Down-regulated expression of cytochrome P450 related genes maybe involved in the development and progression of HCC due to the increased activity of certain carcinogens and maybe the exact mechanism need further study.