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Archive > Volume 30 Issue 3 > 2023,30(3):230-236. DOI:10.3872/j.issn.1007-385X.2023.03.007 Prev Next
Basic Research
Expression of COUP-TF Ⅱ in gastric cancer tissues and its regulation of transcriptional activity of VEGFR3-NRP2 axis in gastric cancer SGC7901 cells
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Abstract:
Objective: To explore the molecular mechanism of COUP-TFⅡ regulating lymphatic metastasis-related factor VEGFR3-NRP2 axis in gastric cancer. Methods: Sixty cases of gastric cancer tissues and paracancerous tissues surgically resected at the Binzhou Medical University Hospital between March 2015 and August 2015 were collected, and the expression of COUP-TFⅡ, VEGFR3 and NRP2 in the tissues was detected by immunohistochemistry and qPCR. Gastric cancer SGC7901 and BGC823 cells were cultured, and COUP-TF Ⅱ overexpression and siRNA-COUP-TF Ⅱ plasmids were constructed and further transfected into SGC7901 cells. The expression of COUP-TFⅡ, VEGFR3 and NRP2 in SGC7901 cells after transfection was detected by WB and qPCR, and the targeting relationship between COUP-TFⅡ and VEGFR3-NRP2 axis was verified by immunoprecipitation (CHIP) and dual-luciferase reporter assay. Results: Immunohistochemistry showed that VEGFR3, NRP2, and COUP-TFⅡ were highly expressed in gastric cancer tissues (all P<0.01). qPCR results showed that the mRNA levels of VEGFR3, NRP2 and COUP-TFⅡ were also highly expressed in gastric cancer tissues compared with paraneoplastic and normal tissues (P<0.05 or P<0.001). WB and qPCR methods showed that compared with the control group, both the mRNA and protein levels of COUP-TFⅡ were significantly higher in the COUP-TFⅡ overexpression group (both P<0.01); while the mRNA and protein levels of COUP-TFⅡ were significantly decreased in the SGC7901 cells of COUP-TFⅡ knockdown group (P<0.05 or P<0.01), and the VEGFR3 and NRP2 mRNA levels were also significantly decreased (both P<0.01). CHIP results showed that the immunoprecipitates of COUP-TFⅡ antibodies in SGC7901 and BGC823 cell lines contained VEGFR3 and NRP2 promoter DNA sequences. The results of dual-luciferase reporter assay showed that COUP-TFⅡ expression level was positively correlated with VEGFR3 and NRP2 levels. Conclusion: COUP-TF Ⅱ has a direct positive regulatory effect on the VEGFR3-NRP2 axis and is highly expressed in gastric cancer. COUP-TFⅡ may serve as a new target for the treatment of gastric cancer.
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