[摘 要] 目的：探究长链非编码RNA00894（LINC00894）调节微小RNA-205-5p（miR-205-5p）/糖蛋白非转移性黑色素瘤蛋白 B（GPNMB）轴对胃癌细胞恶性生物学行为的影响。方法：收集2022年11月至2023年9月在河北医科大学第一医院手术切除的 25例胃癌组织及相应癌旁组织，常规培养BGC823细胞，随机将其分为对照组、sh-NC组、sh-LINC00894组、sh-LINC00894 + anti NC组、sh-LINC00894 + anti-miR-205-5p组，用转染试剂将相应质粒转染至各组细胞中。qPCR法检测各组BGC823细胞和癌组织 中LINC00894、miR-205-5p 和 GPNMB mRNA 表达，双萤光素酶报告基因实验和AGO2-RNA免疫共沉淀验证LINC00894与 miR-205-5P和miR-205-5p与GPNMB间的靶向结合关系。克隆形成实验、EdU染色、划痕愈合实验和Transwell实验分别检测各 组细胞的增殖、迁移和侵袭能力。WB法检测各组细胞中CDK1、MMP-2和MMP-9蛋白的表达。裸鼠移植瘤实验检测敲减 LINC00894对移植瘤生长的影响，免疫组化法检测移植瘤组织中GPNMB蛋白的表达。结果：胃癌组织和细胞中LINC00894、 GPNMB呈高表达，miR-205-5p呈低表达（均P < 0.05）。LINC00894与miR-205-5p和miR-205-5p与GPNMB之间存在靶向结合负 向调控关系（均P < 0.05）。敲减LINC00894可促进BGC823细胞中miR-205-5p表达并抑制GPNMB表达（均P < 0.05），敲减 LINC00894可抑制BGC823细胞的增殖、迁移和侵袭能力，以及抑制CDK1、MMP-2和MMP-9蛋白的表达（均P < 0.05），抑制 miR-205-5p则可逆转此作用（均P < 0.05）。敲减LINC00894可抑制BGC823细胞移植瘤的生长、促进miR-205-5p表达、抑制 GPNMB蛋白表达（均P < 0.05）。结论：在胃癌组织及细胞中LINC00894呈高表达，miR-205-5p呈低表达，敲减LINC00894表达 可调控BGC823细胞中miR-205-5p/GPNMB通路蛋白表达并抑制其恶性生物学行为。

[Abstract] Objective: To investigate the effects of long intergenic non-coding RNA00894 (LINC00894) on the malignant biological behaviors of gastric cancer (GC) cells via regulating the microRNA-205-5p (miR-205-5p)/glycoprotein non-metastatic melanoma protein B (GPNMB) axis. Methods: Twenty-five pairs of gastric cancer tissues and corresponding adjacent tissues were collected from patients undergoing surgical resection at the First Hospital of Hebei Medical University between November 2022 and September 2023. BGC823 cells were routinely cultured and into control, sh-NC, sh-LINC00894, sh-LINC00894 + anti-NC, and sh-LINC00894+anti miR-205-5p groups, followed by corresponding plasmids transfection using transfection reagents. The mRNA expression of LINC00894, miR-205-5p, and GPNMB in BGC823 cells and cancer tissues of each group was detected using qPCR. Dual luciferase reporter gene assay and AGO2-RNA immunoprecipitation were used to verify the targeted binding relationships between LINC00894 and miR-205-5p, as well as between miR-205-5p and GPNMB. Colony formation assay, EdU staining, wound-healing assay, and Transwell assay were performed to detect cell proliferation, migration and invasion abilities. Western blotting was used to detect the protein expression of CDK1, MMP-2 and MMP-9 in each group of cells. A nude mouse xenograft model was established to determine the effect of LINC00894 knockdown on tumor growth, and immunohistochemistry was used to detect the protein expression of GPNMB in xenograft tissues. Results: LINC00894 and GPNMB were significantly upregulated in gastric cancer tissues and cells,while miR-205-5p was downregulated (both P < 0.05). LINC00894 negatively regulated miR-205-5p, while miR-205-5p negatively regulated GPNMB (all P < 0.05). Knocking down LINC00894 promoted miR-205-5p expression and inhibited GPNMB expression in BGC823 cells (all P < 0.05). Additionally, LINC00894 knockdown suppressed the proliferation, migration and invasion abilities of BGC823 cells, as well as the protein expression of CDK1, MMP-2, and MMP-9 (all P < 0.05). Inhibition of miR-205-5p reversed these effects (all P < 0.05). Knocking down LINC00894 inhibited the growth of BGC823 cell-transplanted tumors, promoted miR-205-5p expression, and inhibited GPNMB protein expression (all P < 0.05). Conclusion: LINC00894 is highly expressed in gastric cancer tissues and cells, while miR-205-5p is downregulated. Knockdown of LINC00894 can regulate the expression of miR-205-5p/GPNMB pathway-related proteins in BGC823 cells, thereby inhibiting their malignant biological behaviors.

[基金项目] 河北省2022年政府资助临床医学优秀人才培养项目（No. LS202208）