[摘 要] 目的：探究长链非编码RNA葡萄糖醛酸酶β假基因11（GUSBP11）调节miR-339-5p/小鼠双分钟同源物2（MDM2）轴 对胃癌细胞AGS增殖、迁移和侵袭的影响。方法：收集2023年12月至2024年6月期间在广州中医药大学附属佛山中医院手术 治疗的25例胃癌患者的癌旁组织及癌组织。常规培养胃癌细胞AGS和正常胃黏膜上皮细胞GES-1，用转染试剂将对照质粒和 敲减质粒转染AGS细胞，分为Ctrl组、sh-NC、sh-GUSBP11、sh-GUSBP11 + anti-NC、sh-GUSBP11 + anti-miR-339-5p。qPCR法检 测胃癌组织及各组细胞中GUSBP11、miR-339-5p和MDM2 mRNA的表达；双萤光素酶报告基因实验检测GUSBP11或MDM2与 miR-339-5p间的靶向关系；EdU法检、Transwell小室实验、划痕愈合实验和WB法分别检测各组AGS的增殖、迁移和侵袭能力和 细胞中CDK1、MMP-2、MMP-9蛋白的表达；AGS细胞移植瘤实验检测敲减GUSBP11对移植瘤生长的影响。结果：胃癌组织和 细胞中GUSBP11、MDM2 mRNA均呈高表达（均P < 0.05），miR-339-5p呈低表达（P < 0.05）。GUSBP11与miR-339-5p和MDM2 与miR-339-5p间存在靶向关系。在AGS细胞中敲减GUSBP11可明显抑制MDM2蛋白、促进miR-339-5p的表达而抑制miR-339-5p 则可促进MDM2蛋白表达。敲减GUSBP11可抑制AGS细胞的增殖、迁移和侵袭能力而抑制miR-339-5p则可逆转此作用。敲减 GUSBP11可明显抑制CDK1、MMP-2和MMP-9蛋白的表达而抑制miR-339-5p则可逆转此作用。敲减GUSBP11可明显抑制 AGS细胞移植瘤的生长。结论：GUSBP11在胃癌组织和细胞中呈高表达，敲减GUSBP11表达可能通过调控miR-339-5p/MDM2 轴抑制胃癌细胞的恶性生物学行为。

[Abstract] Objective: To investigate the effect of long non-coding RNA glucuronidase β pseudogene 11 (GUSBP11) regulating miR-339-5p/mouse two-minute homolog 2 (MDM2) axis on the proliferation, migration, and invasion of gastric cancer AGS cells. Methods: Cancerous and adjacent tissues from 25 gastric cancer patients who underwent surgical treatment at Foshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine from December 2023 to June 2024 were collected. Gastric cancer AGS cells and normal gastric mucosal epithelial GES-1 cells were routinely cultured. The control plasmids and knockdown plasmids were transfected into AGS cells using transfection reagents, dividing the cells into Ctrl group, sh-NC group, sh-GUSBP11 group, sh-GUSBP11 + anti-NC group, and sh-GUSBP11 + anti-miR-339-5p group. The mRNA expression of GUSBP11, miR-339-5p, and MDM2 in gastric cancer tissues and cells of each group was detected by qPCR. A dual-luciferase reporter gene assay was used to detect the targeting relationship between GUSBP11 or MDM2 and miR-339-5p. EdU staining, scratch healing assay, and Transwell chamber assay were adopted to assess the proliferation, migration, and invasion abilities of AGS cells, respectively. WB assay was used to measure the protein expression of CDK1, MMP-2, and MMP-9 in AGC cells. The effects of GUSBP11 knockdown on tumor growth were examined through AGS cell xenograft experiments. Results: The mRNA expression of GUSBP11 and MDM2 were significantly upregulated in gastric cancer tissues and cells (both P < 0.05), while miR-339-5p was significantly downregulated (P < 0.05). A targeting relationship was found between GUSBP11 and miR-339-5p, as well as between MDM2 and miR-339-5p. Knockdown of GUSBP11 in AGS cells significantly inhibited MDM2 protein expression and promoted miR-339-5p expression, while inhibition of miR-339-5p promoted MDM2 protein expression. GUSBP11 knockdown significantly inhibited the proliferation, migration, and invasion of AGS cells, while inhibition of miR-339-5p reversed this effect. GUSBP11 knockdown significantly inhibited the protein expression of CDK1, MMP-2, and MMP-9, and inhibition of miR-339-5p reversed this effect. Furthermore, GUSBP11 knockdown significantly inhibited the growth of AGS cell xenografts. Conclusion: GUSBP11 is highly expressed in gastric cancer tissues and cells, and knocking down GUSBP11 expression may inhibit malignant biological behaviors of gastric cancer cells through regulating the miR-339-5p/DM2 axis.

广东省基础与应用基础研究基金（No. 2022A1515220156）