Korean J Physiol Pharmacol.  2025 May;29(3):337-347. 10.4196/kjpp.24.275.

Hyaluronan-mediated motility receptor-mediated aerobic glycolysis enhances stem-like properties and chemoresistance in lung adenocarcinoma

• Yu W
• Shi Y
• Bao X
• Chen X
• Ni Y
• Wang J
• Ye H

• KMID: 2567191
• DOI: http://doi.org/10.4196/kjpp.24.275

Abstract

Lung adenocarcinoma (LUAD) is a global malignancy with significant chemoresistance impacting patient prognosis. The pro-tumorigenic role of hyaluronan-mediated motility receptor (HMMR) in LUAD is recognized. This study was designed to investigate the underlying mechanisms by which HMMR affects chemoresistance in LUAD. Bioinformatics presented the expression patterns of HMMR in LUAD patients and the association between HMMR levels and patient survival, followed by qRT-PCR to verify HMMR expression in LUAD tissues and cells. Further, bioinformatics was leveraged to identify the signaling pathways enriched by HMMR and its relevance to glycolytic genes, we also analyzed changes in the glycolytic activity of LUAD cells by manipulating HMMR expression. Stemness was evaluated through cell aggregation assays and Western blot, and drug responsiveness was gauged using CCK-8 assays, alongside flow cytometry for apoptosis analysis. HMMR was highly expressed in LUAD tissues and cells, and this overexpression correlated with poorer prognoses in patients. GSEA showed that HMMR was notably enriched in the glycolysis and gluconeogenesis pathways, correlating positively with the expression of key glycolytic genes. Cellular experiments confirmed that HMMR knockdown notably suppressed aerobic glycolysis in LUAD cells. Moreover, overexpression of HMMR could further enhance the stemness and cisplatin resistance of LUAD cells by stimulating glycolysis. In brief, this study has validated that high levels of HMMR in LUAD are predictive of poor patient prognosis, and that overexpression of HMMR can catalyze aerobic glycolysis, thus promoting stemness and chemoresistance in LUAD cells. Thus, HMMR could be a target for improving chemosensitivity in LUAD.

Keyword

Adenocarcinoma of lung; Drug resistance, neoplasm; Glycolysis; Hyaluronan receptors; Neoplastic stem cells

Figure

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  Fig. 1 High expression of hyaluronan-mediated motility receptor (HMMR) in lung adenocarcinoma (LUAD) correlates with worsened patient outcomes. (A) TCGA database analysis of HMMR expression in normal lung tissues (N = 59) versus LUAD tissues (N = 539). (B) K-M curve analysis of overall survival time in patients with high/low HMMR expression (N = 1,082/1,084). (C) qRT-PCR detection of HMMR mRNA expression in cancerous and adjacent tissues (N = 10/10). (D) qRT-PCR detection of HMMR mRNA expression in tumor tissues of stage I/II (N = 4) versus stage III/IV (N = 6) LUAD patients. (E) qRT-PCR evaluation of HMMR mRNA levels in normal human lung epithelial cells and LUAD cells (N = 3). Values are presented as mean ± SD. * signifies p < 0.05.

• 

  Fig. 2 High hyaluronan-mediated motility receptor (HMMR) expression enhances the stemness and drug resistance of lung adenocarcinoma (LUAD) cells. (A) si-NC and si-HMMR were transfected into PC9 lung adenocarcinoma cells, and transfection efficiency was confirmed by qRT-PCR (N = 3). (B, C) Spheroid formation detected stemness among different treatment groups (N = 3). More sphere formation represents more stemness of the cell. (D) WB examined the expression of stem cell markers CD44 and CD133. (E) The CCK-8 assay calculated the cisplatin IC50 values for each group (N = 3). Lower IC50 values indicate greater cisplatin sensitivity. (F) Apoptosis was evaluated using flow cytometry (N = 3). Values are presented as mean ± SD. WB, Western blot; PI, propidium iodide. * signifies p < 0.05.

• 

  Fig. 3 Hyaluronan-mediated motility receptor (HMMR) expression mediates glycolytic signaling. (A) GSEA analysis of signaling pathways enriched for genes highly associated with HMMR. (B) Pearson analysis of the correlation between HMMR and key genes of glycolysis (PKM, LDHA, SLC2A1, PDK1, MYC). A larger value means a stronger correlation. (C) WB analysis of LDHA, PDK1, and MYC protein expression in si-NC and si-HMMR groups. (D) ECAR measurement via the Seahorse XF96 Extracellular Flux Analyzer (N = 3). ECAR levels are positively correlated with glycolysis. (E) OCR measurement via the Seahorse XF96 (N = 3). OCR levels are negatively correlated with glycolysis. (F) Glucose consumption measured in cell supernatants of different groups with a specific assay kit (N = 3). (G) Lactate synthesis quantified in the cell groups with a specific assay kit (N = 3). (H) ATP yield evaluated in the cell groups with a specific assay kit (N = 3). Values are presented as mean ± SD. WB, Western blot; ECAR, extracellular acidification rate; OCR, oxygen consumption rate; 2-DG, 2-Deoxy-D-glucos. * signifies p < 0.05.

• 

  Fig. 4 Hyaluronan-mediated motility receptor (HMMR) enhances stemness and drug resistance of lung adenocarcinoma (LUAD) cells by promoting glycolytic metabolism. (A) Transfection of A549 cells with oe-NC and oe-HMMR, with qRT-PCR assessing transfection efficiency (N = 3). (B) Cells were divided into groups as oe-NC + PBS, oe-HMMR + PBS, oe-HMMR + 2-DG, with ECAR analyzed by the Seahorse XF96 (N = 3). (C) OCR measured with the Seahorse XF96 (N = 3). (D) Glucose consumption in supernatants of the groups assessed by a specific assay kit (N = 3). (E) Lactate production in the groups measured by an assay kit (N = 3). (F) ATP production in the groups evaluated with a specific assay kit (N = 3). (G) WB detection of HMMR and glycolysis-related protein (LDHA, PDK1, MYC) expression. (H) Spheroid formation assays for stemness in different groups of cells (N = 3). (I) WB detection of stem cell markers CD44 and CD133 expression. (J) CCK-8 assay for cisplatin IC50 in each group (N = 3). (K) Flow cytometry for apoptosis assessment in cells (N = 3). Values are presented as mean ± SD. PBS, phosphate-buffered saline; 2-DG, 2-Deoxy-D-glucos; ECAR, extracellular acidification rate; OCR, oxygen consumption rate; WB, Western blot; PI, propidium iodide. * signifies p < 0.05.

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