Proteomic analysis of the pro-proliferative role of the protein lysosomal acid glucosylceramidase in hepatocellular carcinoma
10.3760/cma.j.cn113884-20240709-00203
- VernacularTitle:蛋白质组学分析蛋白溶酶体酸性葡糖神经酰胺酶在肝细胞癌中的促增殖作用
- Author:
Tong XIAO
1
;
Hao WANG
;
Hongle LI
Author Information
1. 郑州大学附属肿瘤医院（河南省肿瘤医院）检验科，郑州 450008
- Publication Type:Journal Article
- Keywords:
Carcinoma, hepatocellular;
Proteomics;
Cell proliferation
- From:
Chinese Journal of Hepatobiliary Surgery
2025;31(3):219-225
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To integrate and analyze large-scale proteomic datasets from clinical hepatocellular carcinoma (HCC) samples to identify key oncogenic proteins and investigate their underlying mechanisms.Methods:Based on two publicly reported large-scale proteomic datasets of hepatocellular carcinoma, we identified lysosomal acid glucosylceramidase (GBA) as a differentially expressed protein between tumor and adjacent non-tumor tissues. The Cancer Genome Atlas (TCGA) database was utilized to analyze survival differences between patients with high ( n=118) and low ( n=118) GBA expression. Tumor and adjacent non-tumor tissue samples were collected from eight HCC patients who underwent surgical treatment at Henan Cancer Hospital between March 2021 and May 2021. Among them, six were male and two were female, ranging from 41 to 66 years old, with a median age of 56 years. The human HCC cell line HCC-LM3 was transfected with siRNAs targeting GBA (siGBA-1 and siGBA-2) or a negative control (siNC). Western blot analysis was performed to assess the expression of relevant proteins in HCC tissues and cells. Quantitative proteomic analysis was conducted to identify differentially expressed proteins in HCC cells, and bioinformatics tools were used to construct an interaction network of downregulated proteins. Cell proliferation and colony formation assays were conducted to evaluate the impact of GBA expression on HCC cell growth. Results:GBA was identified as a significantly upregulated protein in HCC tissues across two clinical proteomic datasets. TCGA analysis further revealed that high GBA expression was associated with poor prognosis in HCC patients. Western blot analysis confirmed that the relative expression level of GBA protein in clinical HCC tissues was (0.60±0.35), which was significantly higher than that in adjacent non-tumor tissues (0.26±0.20) ( t=2.84, P=0.025). Quantitative proteomic analysis interaction analysis of GBA knockdown and control cells identified epidermal growth factor receptor (EGFR) as a potential downstream regulatory target of GBA. Western blot validation demonstrated that EGFR protein expression was significantly reduced following GBA knockdown [siNC: (0.92±0.08), siGBA-1: (0.64±0.07), siGBA-2: (0.51±0.07)]. Cell proliferation and colony formation assays showed that GBA knockdown significantly inhibited cell growth and colony-forming ability. The OD450 values on day 4 in control group and GBA knockdown group (siGBA-1 and siGBA-2) were (1.91±0.17), (1.24±0.11), and (1.21±0.04), respectively. The colony numbers were (674.33±6.51), (388.33±7.51), and (360.00±29.00), respectively. The knockown groups were both lower than negative control group and the differences were statistically significant (both P<0.05). Conclusion:GBA is significantly overexpressed in HCC tissues and may promote HCC cell proliferation by regulating EGFR protein expression.
