Abstract
Asparagine-linked glycosylation protein 1 homolog (ALG1) participates in the initial stage of protein N-glycosylation and N-glycosylation has been implicated in the process of hepatocellular carcinoma (HCC) progression. However, whether ALG1 plays a role in human HCC remains unknown. In this study, the expression profile of ALG1 in tumorous and corresponding adjacent non-tumor tissues was analyzed. The relationship of ALG1 expression with clinical features and prognosis of HCC patients was also evaluated using immuno-histochemical method. Here we found ALG1 decreased in HCC tissues compared with adjacent normal liver tissues, which predicted an unfavorable prognosis. Combined with RNA interference, nascent proteome and glycoproteome were determined systematically in Huh7 cell line. Bioinformatics analysis indicated that the differentially expressed proteins participating in the response of ALG1 knockdown were most significantly associated with cell–cell adhesion. Functional studies confirmed that knockdown of ALG1 reduced cell adhesion capacity, and promoted cell migration. Furthermore, down-regulation of H8N2 (on N-glycosite N651) and H5N4S2F1 (on N-glycosite N692) from N-cadherin was identified as a feature of ALG1 knockdown. Our findings revealed that ALG1 controlled the expression of glycosylated N-cadherin and played a role in HCC migration, with implications for prognosis.
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Data Availability
Partial mass spectrometric data and analyzed result datasets have been deposited in iProX (http://www.iprox.org), which is an official member of ProteomeXchange Consortium. The project ID is IPX0003587000.
Code Availability
Not applicable.
Abbreviations
- ALG1:
-
Asparagine-linked glycosylation protein 1 homolog
- HCC:
-
Hepatocellular carcinoma
- ER:
-
Endoplasmic reticulum
- CDG:
-
Congenital disorders of glycosylation
- RT-qPCR:
-
Real-time quantitative PCR
- HRP:
-
Horseradish peroxidase
- DMEM:
-
Dulbecco's modified Eagle's medium
- FBS:
-
Fetal bovine serum
- siRNA:
-
Small interfereing RNA
- AHA:
-
Azidohomoalanine
- GO:
-
Gene ontology
- BP:
-
Biological process
- H:
-
Hexose
- N :
-
N-Acetylhexosamine
- S:
-
Sialic acid
- F:
-
Fucose
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Acknowledgements
We thank openbiox community and Hiplot team (https://hiplot.com.cn) for providing technical assistance and valuable tools for data analysis and visualization. The authors thank Shu Zhang from Zhongshan Hospital, Fudan University, Shanghai, China, for her generous support. We acknowledge the financial support of the National Key Research and Development Program of China (2017YFA0505100) and NSF of China (Grants 21974025 and 82121004) for this work.
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Conceptualization: XC. Methodology: YS and PM. Visualization: ZC. Investigation: GY, JY, XZ. Software: CL. Data curation: LZ. Writing—original draft: HS. Writing—reviewing and editing: HL.
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The studies involving human participants were reviewed and approved by the Research Ethics Committee of First Affiliated Hospital of Guangxi Medical University. Approval number was 2019(KY-E-086). The privacy rights of human subjects were also observed. No animal experimentation was performed.
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Cao, X., Shao, Y., Meng, P. et al. Nascent Proteome and Glycoproteome Reveal the Inhibition Role of ALG1 in Hepatocellular Carcinoma Cell Migration. Phenomics 2, 230–241 (2022). https://doi.org/10.1007/s43657-022-00050-5
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DOI: https://doi.org/10.1007/s43657-022-00050-5