Role and mechanism of sarcosine dehydrogenase in the progression of gallbladder cancer through chemokine pathways

Abstract

Background: Sarcosine dehydrogenase (SARDH) and C-X-C motif chemokine ligand 1 (CXCL1) have been identified as potential tumor regulators, with growing evidence linking them to cancer progression. However, their specific roles, regulatory mechanisms, and influence on key signaling pathways remain unclear.

Aim: To investigate the regulatory mechanisms of SARDH and CXCL1 in cancer cells and their impact on key signaling pathways.

Methods: Real-time quantitative polymerase chain reaction and western blot analyses were used to assess the expression levels of SARDH and CXCL1 and their effects on protein kinase B (Akt) and extracellular signal-regulated kinase (ERK) signaling pathways. Gene overexpression was induced using an expression vector, while gene silencing was achieved using short hairpin RNA and small interfering RNA. CCK-8, migration, and invasion assays were used to evaluate the impact of gene suppression and overexpression on cancer cell proliferation, migration, and invasion.

Results: SARDH silencing significantly enhanced cancer cell proliferation, whereas its overexpression suppressed proliferation in the early stages of the experiment. CXCL1 silencing reduced cancer cell migration and invasion. SARDH overexpression inhibited cell migration, invasion, and adhesion while increasing apoptosis. Conversely, SARDH silencing reversed these effects. Furthermore, simultaneous silencing of SARDH and CXCL1 strongly activated the Akt and ERK signaling pathways, indicating the potential role of these pathways in regulating cellular functions influenced by these genes.

Conclusion: This study revealed that SARDH and CXCL1 regulate cancer cell growth, migration, and invasion through Akt and ERK signaling pathways, highlighting their potential as therapeutic targets for cancer treatment.

Keywords: Chemokines; Extracellular signal-regulated kinase; Gallbladder cancer; Protein kinase B; Sarcosine dehydrogenase.

Figure 1

Sarcosine dehydrogenase expression in gallbladder cancer tissues and cell lines. A: Real-time quantitative polymerase chain reaction (RT-PCR) analysis of the expression of sarcosine dehydrogenase (SARDH) in gallbladder cancer tissues; B: RT-PCR analysis of SARDH expression in various gallbladder cancer cell lines. SARDH: Sarcosine dehydrogenase.

Figure 2

Effect of sarcosine dehydrogenase on gallbladder cancer cell proliferation. A: Real-time quantitative polymerase chain reaction (RT-PCR) analysis confirms the efficiency of sarcosine dehydrogenase (SARDH) knockdown; B: RT-PCR analysis confirming the efficiency of SARDH overexpression; C: CCK-8 assay showing that SARDH knockdown promotes gallbladder cancer cell proliferation; D: CCK-8 assay showing that SARDH overexpression inhibits gallbladder cancer cell proliferation. SARDH: Sarcosine dehydrogenase.

Figure 3

Effects of sarcosine dehydrogenase on the migration, invasion, adhesion, and apoptosis of gallbladder cancer cells. A: Sarcosine dehydrogenase (SARDH) overexpression inhibited the migration and invasion of gallbladder cancer cells; B: SARDH silencing promoted the migration and invasion of gallbladder cancer cells; C: SARDH overexpression inhibited the adhesion of gallbladder cancer cells; D: SARDH silencing promoted cell adhesion of gallbladder cancer cells; E: Flow cytometry analysis showed that SARDH overexpression promoted the apoptosis of gallbladder cancer cells; F: Flow detection of SARDH silencing inhibited the apoptosis of gallbladder carcinoma cells. SARDH: Sarcosine dehydrogenase.

Figure 4

Interaction between sarcosine dehydrogenase and C-X-C motif chemokine ligand 1 in gallbladder cancer and its impact on signaling pathways. A: Real-time quantitative polymerase chain reaction (RT-PCR) detection of the interaction between sarcosine dehydrogenase (SARDH) and C-X-C motif chemokine ligand 1 (CXCL1); B: RT-PCR detection of CXCL1 expression in gallbladder cancer; C: Western blot analysis showing activation of protein kinase B (Akt) and extracellular signal-regulated kinase (ERK)1/ERK2 signaling pathways through the SARDH/CXCL1 axis in BGC-SD cells; D: Silencing SARDH and CXCL1 markedly increased the activation levels of p-Akt and p-ERK in cancer cells. SARDH: Sarcosine dehydrogenase; Akt: Protein kinase B; ERK: Extracellular signal-regulated kinase.

Figure 5

Effects of C-X-C motif chemokine ligand 1 silencing on the proliferation, migration, and invasion of gallbladder cancer cells. A: Real-time quantitative polymerase chain reaction analysis confirmed the efficiency of C-X-C motif chemokine ligand 1 (CXCL1) silencing; B: CCK-8 assay showed that CXCL1 silencing inhibited gallbladder cancer cell proliferation; C: CXCL1 silencing inhibited gallbladder cancer cell migration and invasion. CXCL1: C-X-C motif chemokine ligand 1.

Figure 6

Synergistic effects of sarcosine dehydrogenase and C-X-C motif chemokine ligand 1 regulation on the function of gallbladder cancer cells. A: Gallbladder cancer cell proliferation was detected after sarcosine dehydrogenase (SARDH) overexpression and C-X-C motif chemokine ligand 1 (CXCL1) silencing; B: Gallbladder cancer cell migration and invasion were detected after SARDH overexpression and CXCL1 silencing; C: Gallbladder cancer cell proliferation was detected after simultaneous overexpression of SARDH and CXCL1; D: Gallbladder cancer cell proliferation was detected after overexpression of both SARDH and CXCL1; E: Western blot analysis showed that interference with the SARDH/CXCL1 axis inhibited the activation of protein kinase B and extracellular signal-regulated kinase (ERK)1/ERK2 signaling pathways in BGC-SD. CXCL1: C-X-C motif chemokine ligand 1; SARDH: Sarcosine dehydrogenase; Akt: Protein kinase B; ERK: Extracellular signal-regulated kinase.