[摘 要] 目的：探讨KH型剪切调节蛋白（KHSRP）靶向调控JAK1/STAT3信号轴对食管胃结合部腺癌（AEG）细胞增殖、迁移 和侵袭及移植瘤生长与肺转移的影响。方法：收集2017年1月至2018年12月间在淮河医院确诊的64例AEG组织和癌旁组织 标本及临床资料，采用免疫组化法观察AEG组织和癌旁组织中KHSRP的表达水平。qPCR法检测AEG细胞OE-19、TE-7、BIC-1、 FLO-1、SK-GT-4、BE-3与正常食管上皮细胞Het-1A中KHSRP的表达差异。通过慢病毒载体对KHSRP进行敲减和过表达处理， 分别转染 OE-19 与 TE-7 细胞、FLO-1 与 SK-GT-4 细胞，实验分为 sh-NC 组、sh-KHSRP 组和 Vector 组、KHSRP 过表达组（KHSRP 组）。采用qPCR法检测敲减或过表达效率，CCK-8法、Transwell小室法分别检测敲减或过表达KHSRP对AEG细胞增殖、迁移和 侵袭的影响。构建小鼠异种移植瘤和肺转移模型，观察KHSRP对移植瘤体内生长和转移的作用。WB法验证KHSRP靶向JAK/ STAT信号通路。细胞拯救实验验证KHSRP是否通过调节JAK1/STAT3信号通路促进AEG细胞的恶性进程。结果：与癌旁组织 相比，AEG组织中KHSRP表达水平显著增高（P < 0.05或P < 0.01）。细胞功能实验分析显示，KHSRP过表达在体外均显著促进 AEG细胞增殖、迁移和侵袭（P < 0.05或P < 0.01）。动物实验结果显示，KHSRP在裸鼠体内具有促进AEG细胞移植瘤生长与肺 转移的作用（P < 0.05 或 P < 0.01）。在敲减 KHSRP 后，JAK/STAT 信号通路中 JAK1、STAT3 磷酸化水平均明显降低，过表达 KHSRP后情况则均反之（P < 0.05或P < 0.01）。细胞拯救实验显示，KHSRP可以逆转敲减JAK1/STAT3对细胞增殖、迁移和侵袭 的抑制作用（P < 0.05或P < 0.01）。结论：KHSRP通过激活JAK1/STAT3信号通路调控AEG细胞转移的恶性进程，KHSRP有望 成为AEG临床治疗的潜在靶点。

[Abstract] Objective: To investigate the effects of KH-type splicing regulatory protein (KHSRP) targeting and regulating JAK1/ STAT3 signaling axis on the proliferation, migration and invasion of the adenocarcinoma of esophagogastric junction (AEG) cells, as well as the growth of transplanted tumors and lung metastasis. Methods: A total of 64 pairs of AEG tissue and adjacent normal tissue samples, along with clinical data from patients diagnosed at Huaihe Hospital from January 2017 to December 2018 were collected. The expression level of KHSRP in AEG tissues and adjacent normal tissues was observed using immunohistochemical staining. The differential expression of KHSRP in AEG cells (OE-19, TE-7, BIC-1, FLO-1, SK-GT-4, BE-3) and normal esophageal epithelial cells (Het-1A) was detected by qPCR. Lentiviral vectors were used to knockdown and overexpress KHSRP in OE-19, TE-7, FLO-1, and SK-GT-4 cells. The experiment was divided into the following groups:sh-NC group, sh-KHSRP group, Vector group, and KHSRP overexpression group (KHSRP group). The knockdown or overexpression efficiency was detected by qPCR, and the effects of KHSRP knockdown or overexpression on AEG cell proliferation, migration and invasion were evaluated using CCK-8 and Transwell assays, respectively. A mouse xenograft and lung metastasis model was established to observe the effects of KHSRP on tumor growth and metastasis in vivo. The targeted regulation of JAK/STAT signaling pathway by KHSRP was verified by Western blotting. A rescue experiment was conducted to verify whether KHSRP promoted malignant progression of AEG cells through the JAK1/STAT3 signaling pathway. Results: Compared with adjacent normal tissues, the expression level of KHSRP in AEG tissues was significantly increased (P < 0.05 or P < 0.01). Cell function experiments showed that KHSRP overexpression significantly promoted AEG cell proliferation, migration, and invasion in vitro (P < 0.05 or P < 0.01). In vivo animal experiments showed that KHSRP promoted AEG cell xenograft tumor growth and lung metastasis in nude mice (P < 0.05 or P < 0.01). After KHSRP knockdown, the phosphorylation levels of JAK1 and STAT3 in the JAK/STAT signaling pathway were significantly reduced, while overexpression of KHSRP led to the opposite results (P < 0.05 or P < 0.01). Rescue experiment showed that KHSRP could reverse the inhibition of cell proliferation, migration, and invasion caused by JAK1/STAT3 knockdown (P < 0.05 or P < 0.01). Conclusion: KHSRP regulates the malignant progression of AEG cells by activating JAK1/STAT3 signaling axis. KHSRP may become a potential target for the clinical treatment of AEG.

河南省教育厅资助项目（No. 24A320002，No. 25B320018，No. 25B320022）；河南省科技厅指导项目（No. 232102310007，No. 242102310184， No. 242102310197）