title: Quantitative evaluation of aldo-keto reductase expression in hepatocellular carcinoma (HCC) cell lines.
abstract: The involvement of aldo-keto reductases (AKRs) in tumorigenesis is widely reported, but their roles in the pathological process are not generally recognized due to inconsistent measurements of their expression. To overcome this problem, we simultaneously employed real-time PCR to examine gene expression and multiple reaction monitoring (MRM) of mass spectrometry (MS) to examine the protein expression of AKRs in five different hepatic cell lines. These include one relatively normal hepatic cell line, L-02, and four hepatocellular carcinoma (HCC) cell lines, HepG2, HuH7, BEL7402 and SMMC7721. The results of real-time PCR showed that expression of genes encoding the AKR1C family members rather than AKR1A and AKR1B was associated with tumor, and most of genes encoding AKRs were highly expressed in HuH7. Similar observations were obtained through MRM. Different from HuH7, the protein abundance of AKR1A and AKR1B was relatively consistent among the other four hepatic cell lines, while protein expression of AKR1C varied significantly compared to L-02. Therefore, we conclude that the abundant distribution of AKR1C proteins is likely to be associated with liver tumorigenesis, and the AKR expression status in HuH7 is completely different from other liver cancer cell lines. This study, for the first time, provided both overall and quantitative information regarding the expression of AKRs at both mRNA and protein levels in hepatic cell lines. Our observations put the previous use of AKRs as a biomarker into question since it is only consistent with our data from HuH7. Furthermore, the data presented herein demonstrated that quantitative evaluation and comparisons within a protein family at both mRNA and protein levels were feasible using current techniques.
authors.affiliation: CAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Yang
authors.forename: Ju
authors.initials: J
authors.lastname: Zhang
authors.forename: Shenyan
authors.initials: S
authors.lastname: Zhang
authors.forename: Weiwei
authors.initials: W
authors.lastname: Dong
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Lou
authors.forename: Siqi
authors.initials: S
authors.lastname: Liu
mesh_terms: D000429:Alcohol Oxidoreductases / Q000032:analysis* / Q000235:genetics; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D006528:Carcinoma, Hepatocellular / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics; D045744:Cell Line, Tumor; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008113:Liver Neoplasms / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D059467:Transcriptome
chemical_list: D012333:RNA, Messenger; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase
keywords: Aldo–keto reductase; HCC; Multiple reaction monitoring; Quantitative analysis; Real-time PCR