miR-375靶向SHOX2调控人食管鳞癌细胞侵袭迁移的初步研究

目的 探讨miR-375/SHOX2轴对食管鳞癌细胞侵袭迁移能力的影响，为食管鳞癌的靶向治疗寻找潜在新靶点。方法 选取对数生长期的人食管鳞癌细胞（kyse-70、kyse-30），分别过表达以及干扰miR-375和SHOX2基因，采用克隆形成、MTT、划痕、Transwell等体外细胞功能学实验检测miR-375、SHOX2对食管鳞癌细胞增殖、迁移和侵袭能力的影响，通过拯救实验验证miR-375对SHOX2的靶向调控作用。结果 与对照组的kyse-70、kyse-30细胞相比，过表达miR-375或干扰SHOX2基因后的细胞增殖率、克隆形成率和Transwell细胞穿透数均降低，差异均有统计学意义（P＜0.01）；与对照组的kyse-70、kyse-30细胞相比，干扰miR-375或过表达SHOX2后的细胞增殖率、克隆形成率和Transwell细胞穿透数均升高，差异均有统计学意义（P＜0.01）；miR-375过表达后的食管鳞癌细胞SHOX2基因表达减少，而抑制miR-375后SHOX2基因表达增强。拯救实验结果显示，共转染miR-375和SHOX2后细胞增殖和侵袭能力较对照组又有所增强。结论 miR-375抑制食管鳞癌细胞增殖和侵袭；SHOX2促进食管鳞癌细胞增殖和侵袭。miR-375靶向负调控SHOX2表达，对食管鳞癌细胞的增殖和侵袭起调控作用。

miR-375 targeting SHOX2 in regulation of the invasion and migration of esophageal squamous carcinoma cancer

Objective To explore the influence of miR-375/SHOX2 axis in invasion and migration ability of esophageal squamous cell carcinomas （ESCC） by using cells in vitro， for searching potential new targets in therapy for ESCC. Methods Human ESCC cells，including kyse-70 and kyse-30 in the logarithmic growth phase were used in this study. The colony formation assay，Wound healing assay，MTT assay and Transwell assay were used to testify the effect of over-expression or down-expression of miR-375 or SHOX2 gene on ESCC cells. Then， rescue experiment was used to test the interaction between miR-375 and SHOX2. Results Compared with the negative control group， the proliferation rate， clone forming rate and penetrating number by Transwell were significantly reduced in cells after being over-expressed miR-375 or down-expressed SHOX2 （P＜0.01）. In contrast， compared with the negative control group， the proliferation rate， clone forming rate and penetrating number by Transwell were significantly increased in cells after being down-expressed miR-375 or over-expressed SHOX2 （P＜0.01）. SHOX2 expression was reduced by the over-expression of miR-375， while induced by the down-expression of miR-375 on ESCC cells. Rescue experiment showed co-transfection with miR-375 and SHOX2 reversed the low ability of proliferation and migration induced by over-expression of miR-375. Conclusion miR-375 could inhibit the ability of proliferation， invasion and metastasis of ESCC cells. SHOX2 could promote the ability of proliferation， invasion and metastasis of ESCC cells. The expression of SHOX2 could be regulated by miR-375 for proliferation， invasion and metastasis of ESCC cells. Therefore， miR-375 and SHOX2 could be new potential targets for drug treatment in ESCC.