Identification of ALDH6A1 as a Potential Molecular Signature in Hepatocellular Carcinoma via Quantitative Profiling of the Mitochondrial Proteome

Heon Shin, Hyun Jeong Cha, Min Jung Lee, Keun Na, Donha Park, Chae Yeon Kim, Dai Hoon Han, Hoguen Kim, Young Ki Paik

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8 Citations (Scopus)


Various liver diseases, including hepatocellular carcinoma (HCC), have been linked to mitochondrial dysfunction, reduction of reactive oxygen species (ROS), and elevation of nitric oxide (NO). In this study, we subjected the human liver mitochondrial proteome to extensive quantitative proteomic profiling analysis and molecular characterization to identify potential signatures indicative of cancer cell growth and progression. Sequential proteomic analysis identified 2452 mitochondrial proteins, of which 1464 and 2010 were classified as nontumor and tumor (HCC) mitochondrial proteins, respectively, with 1022 overlaps. Further metabolic mapping of the HCC mitochondrial proteins narrowed our biological characterization to four proteins, namely, ALDH4A1, LRPPRC, ATP5C1, and ALDH6A1. The latter protein, a mitochondrial methylmalonate semialdehyde dehydrogenase (ALDH6A1), was most strongly suppressed in HCC tumor regions (∼10-fold decrease) in contrast to LRPPRC (∼6-fold increase) and was predicted to be present in plasma. Accordingly, we selected ALDH6A1 for functional analysis and engineered Hep3B cells to overexpress this protein, called ALDH6A1-O/E cells. Since ALDH6A1 is predicted to be involved in mitochondrial respiration, we assessed changes in the levels of NO and ROS in the overexpressed cell lines. Surprisingly, in ALDH6A1-O/E cells, NO was decreased nearly 50% but ROS was increased at a similar level, while the former was restored by treatment with S-nitroso-N-acetyl-penicillamine. The lactate levels were also decreased relative to control cells. Propidium iodide and Rhodamine-123 staining suggested that the decrease in NO and increase in ROS in ALDH6A1-O/E cells could be caused by depolarization of the mitochondrial membrane potential (Δψ). Taken together, our results suggest that hepatic neoplastic transformation appears to suppress the expression of ALDH6A1, which is accompanied by a respective increase and decrease in NO and ROS in cancer cells. Given the close link between ALDH6A1 suppression and abnormal cancer cell growth, this protein may serve as a potential molecular signature or biomarker of hepatocarcinogenesis and treatment responses.

Original languageEnglish
Pages (from-to)1684-1695
Number of pages12
JournalJournal of Proteome Research
Issue number4
Publication statusPublished - 2020 Apr 3

Bibliographical note

Funding Information:
All clinical samples were obtained from the archives of the Department of Pathology, Yonsei University (Seoul, Korea), and the Liver Cancer Specimen Bank of the National Research Resource Bank Program affiliated with the Korea Research Foundation of the Ministry of Science and Technology. All diagnoses of HCC ( n = 25) ( Table S1 , Supporting Information) were made by pathologists at Severance Hospital, Yonsei University. Twenty-five cases of HCC were included. The mean age (± standard deviation [SD]) and male: female ratio of each group was 54.6 ± 12.5 and 22:3, respectively. This research was approved by the Institutional Review Board (IRB, 4-2015-0474) of the Yonsei University College of Medicine.

Funding Information:
This work was supported by grants from the Korean Ministry of Health and Welfare (HI160274 to Y.-K.P.), the Liver Cancer Specimen Bank funded by the National Research Foundation (2015M3A9B8028339 to H.K.), the Faculty Research Grant of Yonsei University College of Medicine (6-2016-0122 to D.H.H.), and a Severance Surgeon’s Alumni Research Grant (2016-02 to D.H.H.). We thank the staff members of the Yonsei Proteome Research Center for their assistance with this project. We also thank the Severance Hospital Gene Bank for kindly supplying the tumor tissues and materials used in this work.

Publisher Copyright:
Copyright © 2020 American Chemical Society.

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Chemistry(all)


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