[摘 要] 目的：探究纺锤体蛋白1（SPIN1）促进胃腺癌细胞迁移与侵袭的分子机制。方法: 通过TCGA数据库数据分析胃腺 癌组织中SPIN1 mRNA表达与上皮间质转化（EMT）评分、血管生成评分间的相关性。收集2018年8月至2021年11月期间山东 第一医科大学附属省立医院手术切除的52例胃腺癌患者的癌组织制成组织芯片，每例均包含胃腺癌组织、对应癌旁组织及淋巴 结转移灶，通过免疫组织化学法检测胃腺癌组织中 SPIN1 与 STAT3 的蛋白表达水平及相关性。通过 Transwell 实验研究干扰 SPIN1对胃腺癌细胞侵袭与迁移的影响。使用GEPIA2网站分析SPIN1基因与Janus-激酶/信号转导和转录激活因子（JAK/STAT） 通路相关因子在胃腺癌中的表达相关性。通过qPCR法、WB法检测干扰SPIN1后JAK/STAT通路相关mRNA和蛋白的表达变 化。结果: TCGA数据库数据分析结果显示，SPIN1表达与EMT评分和血管生成评分呈正相关（均 P < 0.05）。SPIN1与STAT3 在胃腺癌组织和淋巴结转移灶中表达升高（均 P < 0.05），在癌旁胃黏膜组织中阴性表达。SPIN1与STAT3的表达显著正相关 （P < 0.05）。干扰SPIN1后胃腺癌细胞的迁移、侵袭能力明显降低（P < 0.05 或 P < 0.01）。GEPIA2网站分析结果显示，SPIN1基 因与JAK1、JAK2、STAT1、STAT2及STAT3表达均呈显著正相关（均P < 0.05）。干扰SPIN1后JAK2、STAT3的mRNA水平下降， 而JAK1、STAT1、STAT2的mRNA水平变化不明显。WB法实验结果表明，干扰SPIN1后JAK2、STAT3、p-JAK2及p-STAT3 的蛋 白表达均显著降低（均P < 0.01），过表达SPIN1后JAK2、STAT3、p-JAK2及p-STAT3的蛋白表达均显著升高（均P < 0.01）。结 论: SPIN1可通过参与JAK2/STAT3信号通路促进胃腺癌细胞迁移与侵袭。

[Abstract] Objective: To explore the molecular mechanism of spindlin1 (SPIN1) in promoting the migration and invasion of gastric adenocarcinoma cells. Methods: The correlations between SPIN1 mRNA expression and epithelial-mesenchymal transition (EMT) score and angiogenesis fraction in gastric adenocarcinoma tissues were analyzed by TCGA database. Tissue microarrays were constructed from a total of 52 surgically resected gastric adenocarcinoma specimens collected in Affiliated Provincial Hospital of Shandong First Medical University between August 2018 and November 2021. Each case included matched samples of gastric adenocarcinoma tissue, adjacent non?tumor tissue, and lymph node metastasis. The expression levels and correlation of SPIN1 and STAT3 in the gastric adenocarcinoma tissue samples were subsequently detected using immunohistochemistry. The effect of interfering with SPIN1 on the invasion and migration of gastric adenocarcinoma cells was investigated by Transwell chamber experiment. The GEPIA2 website was used to analyze the expression correlation between the SPIN1 gene and Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway-related factor in gastric adenocarcinoma. Subsequently, quantitative real-time polymerase chain reaction (qPCR) and Western blotting (WB) were employed to detect changes in the expression levels of JAK/STAT pathway-related mRNAs and proteins following SPIN1 interference. Results: Analysis of TCGA data showed that SPIN1 expression was positively correlated with both EMT and angiogenesis scores (both P < 0.05). Elevated expressions of SPIN1 and STAT3 were observed in gastric adenocarcinoma tissues and lymph node metastases (both P < 0.05) while they were negatively expressed in adjacent gastric mucosal tissues. A statistically significant positive correlation was found between SPIN1 and STAT3 expression (P < 0.05). Knockdown of SPIN1 significantly reduced the migratory and invasive capacities of gastric adenocarcinoma cells (P < 0.05 or P < 0.01). Furthermore, GEPIA2 analysis demonstrated significant positive correlations between the SPIN1 gene and the expression levels of JAK1, JAK2, STAT1, STAT2, and STAT3 (all P < 0.05). SPIN1 interference specifically reduced mRNA expressions of JAK2 and STAT3, while no significant changes were detected in JAK1, STAT1, or STAT2 mRNA levels. Western blotting confirmed that SPIN1 knockdown significantly decreased protein expression levels of JAK2, STAT3, phosphorylated JAK2 (p-JAK2), and phosphorylated STAT3 (p-STAT3) (all P < 0.01), whereas SPIN1 overexpression significantly upregulated the expressions of these proteins (all P < 0.01). Conclusion: SPIN1 can promote the migration and invasion of gastric adenocarcinoma cells by participating in the JAK2/STAT3 signaling pathway.

[基金项目] 国家自然科学基金青年基金（82103490）；山东省自然科学基金面上项目（ZR2021MH382）