[摘要] 目的：探讨miR-1269a 在食管癌组织中的表达及其对KYSE30 细胞恶性生物学行为的影响，并研究其可能的作用机制。方法：选取90 例在河北医科大学第四医院通过手术切除的食管癌组织标本，并收集正常食管永生化上皮细胞和食管癌细胞系，应用qPCR 实验检测癌组织和细胞系miR-1269a 的表达水平。将miR-1269a 的mimics 和inhibitor 分别转染食管癌细胞KYSE30 后，用MTS、Transwell 和克隆形成实验分别检测miR-1269a 对KYSE30 细胞增殖、侵袭、迁移和克隆形成能力的影响。通过生物信息学软件预测miR-1269a 的靶基因，并通过WB实验和双荧光素酶报告基因实验验证miR-1269a 对靶基因的调控作用。转染SOX6 质粒后，采用MTS、Transwell 和克隆形成实验分别检测SOX6 对KYSE30 细胞的增殖、侵袭、迁移和克隆形成能力的影响，并利用回复实验进一步验证结果。结果：食管癌组织中miR-1269a 的表达水平显著高于癌旁组织（P<0.05）；与正常食管上皮细胞相比，食管癌细胞系中miR-1269a 的表达水平显著升高（P<0.05 或P<0.01）。miR-1269a mimics 转染组KYSE30 细胞的增殖、侵袭、迁移和克隆形成能力显著高于mimics NC组（P<0.05 或P<0.01）；inhibitor 转染组KYSE30 细胞的增殖、侵袭、迁移和克隆形成能力显著低于inhibitor NC组（P<0.05 或P<0.01）。miR-1269a 可与SOX6 的3’UTR区相结合，且过表达miR-1269a 后，KYSE30细胞的SOX6 表达水平和荧光素酶报告基因的活性均显著降低（P<0.05）。回复实验表明，高表达miR-1269a 可以促进食管癌细胞增殖、侵袭和迁移（P<0.05 或P<0.01），同时高表达SOX6 后，miR-1269a 的促进作用出现部分逆转。结论：miR-1269a 在食管癌组织和细胞系中表达上调，能够促进KYSE30 细胞的增殖、迁移、侵袭和克隆形成等恶性生物学行为，其机制可能是通过抑制靶基因SOX6 实现的。

[Abstract] Objective: To investigate the expression of miR-1269a in esophageal squamous cell carcinoma (ESCC) tissues and its effect on the malignant biological behaviors of ESCC KYSE30 cells, as well as to explore the underlying mechanism. Methods: Ninety specimens of ESCC tissues and adjacent para-cancerous tissues were obtained from patients underwent surgery in Fourth Hospital, Hebei Medical University. In addition, normal esophageal immortalized epithelial cells and esophageal cancer cell lines were also collected.The expression level of miR-1269a in above mentioned tissues and cell lines was examined by Real-time fluorescent quantitative PCR.After being transfected with miR-1269a mimics and inhibitors, the effects of miR-1269a on proliferation, migration, invasion and colony formation of KYSE30 cells were detected by MTS, Transwell and colony formation assay, respectively. The bioinformatics tool was used to predict the possible target genes of miR-1269a. Then the regulation effect of miR-1269a on target gene expression was validated by WB and Dual-luciferase reporter assay. After being transfected with SOX6 plasmid, the effects of SOX6 on the proliferation, migration,invasion and colony formation of KYSE30 cells were detected by MTS, Transwell and colony formation assay, respectively. At last, rescue assay was used to confirm the results. Results: The expression level of miR-1269a in ESCC tissues was significantly higher than that in adjacent para-cancerous tissues (P<0.05), and the expression level of miR-1269a in ESCC cell lines was significantly elevated compared with the normal epithelial cells (P<0.05 or P<0.01). The capacities of proliferation, invasion, migration and colony formation of KYSE30 cells in miR-1269a mimics transfection group were obviously higher than those in mimics NC group, while those abilities in miR-1269a inhibitor transfection group were significantly lower than those in inhibitor NC group (P<0.05 or P<0.01). Bioinformatics analysis showed that miR-1269a could combine with 3’UTR region at SOX6 gene; and after miR-1269a over-expression, the expression level of SOX6 and luciferase activity in KYSE30 cells were significantly reduced (P<0.05). Rescue assay showed that miR-1269a over-expression could promote the proliferation, invasion and migration of KYSE 30 cells, while simultaneous transfection of SOX6 could partially reverse the promotion effect of miR-1269a mimics. Conclusion: The expression level of miR-1269a in ESCC tissues and cell lines is significantly increased, and it could enhance proliferation, migration, invasion and colony formation of KYSE30 cell line. And its mechanism may be related to the suppression of its target gene SOX6.

国家自然科学基金资助项目（No.81772550）；河北省科技计划基金资助项目（No.162777138）