摘要
目的探讨miR-125b-5p对子宫内膜癌细胞增殖、迁移和侵袭的影响及其机制。方法采用荧光定量PCR检测子宫内膜癌组织中miR-125b-5p的表达,免疫组织化学染色法检测子宫内膜癌组织中STAT3、IL-6/10的表达;双荧光素酶报告基因检测和Western blot检测验证miR-125b-5p和STAT3的靶向调控关系;荧光定量PCR和Western blot检测miR-125b-5p过表达或干扰STAT3表达对子宫内膜癌HEC-1B细胞中IL-6/10 mRNA和蛋白表达的影响;CCK-8法、Transwell小室分别检测miR-125b-5p过表达或干扰STAT3表达对HEC-1B细胞的增殖、迁移和侵袭的影响。结果与癌旁正常组织相比,子宫内膜癌组织中miR-125b-5p表达明显降低,而STAT3、IL-6/10蛋白的表达水平明显升高(P<0.05)。子宫内膜癌中STAT3表达水平与肿瘤大小、淋巴结转移、组织学分级以及TNM分期呈显著正相关(P<0.05),而miR-125b-5p表达与肿瘤大小、淋巴结转移、组织学分级以及TNM分期呈显著负相关(P<0.05)。STAT3是miR-125b-5p的潜在靶基因,miR-125b-5p可负向调控STAT3表达。miR-125b-5p过表达后,HEC-1B细胞中IL-6/10 mRNA和蛋白表达水平以及细胞增殖、迁移、侵袭能力均明显降低;干扰STAT3表达后,HEC-1B细胞中IL-6/10表达及细胞增殖、迁移、侵袭能力与miR-125b-5p过表达的结果相一致。结论miR-125b-5p可通过靶向STAT3调控IL-6/10的表达抑制子宫内膜癌细胞的增殖、迁移和侵袭。
To investigate the effect of miR-125b-5p on the proliferation, migration and invasion of endometrial cancer cells and its mechanism, the expression of miR-125b-5p in endometrial cancer tissues was detected by fluorescent quantitative PCR, and the expression of STAT3 and IL-6/10 in endometrial cancer tissues was detected by immunohistochemical staining. The targeting regulation relationship between STAT3 and miR- 125b-5p was verified by double luciferase reporter gene assay and Western blotting;the expression of IL-6 / 10 mRNA and proteins in HEC-1B cells were detected by fluorescence quantitative PCR and Western blotting, respectively. The effects of miR-125b-5p over-expression or STAT3 interference on the proliferation, migration and invasion of HEC-1B cells were detected by CCK-8 method and Trans well chamber test, respectively. Data showed that miR - 125b-5p expression in endometrial carcinoma was significant lower, while the expression levels of STAT3 and IL-6/10 protein were significant higher, as compared with adjacent normal tissues (P<0.05). The expression of STAT3 was positively correlated with tumor size, lymph node metastasis, histological grade and TNM stage (P<0.05),while the expression of miR-125b-5p was negatively correlated with tumor size, lymph node metastasis,histological grade and TNM stage (P<0.05). STAT3 was a potential target gene of miR-125b-5p, and could be negatively regulated by miR-125b-5p. Both miR-125b-5p over-expression and STAT3 interfere could significantly down-regulate the expression of IL-6 / 10 and the proliferation, migration and invasiveness of HEC-1B cells. In conclusion, miR-125b-5 can inhibit the proliferation, migration and invasion of endometrial cancer cells by targeting STAT3 and regulating the expression of IL-6/10.
作者
肖庆
刘莉
胡雅君
姜柳
李金丽
邓小艳
XIAO Qing;LIU Li;HU Yajun;JIANG Liu;LI Jinli;DENG Xiaoyan(Department of Reproductive Medicine,Wuhan First Hospital,Wuhan 430030,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2019年第7期553-560,共8页
Immunological Journal
基金
武汉市卫计委课题(WX14C04)