Xu Hui, Jiang Jialong, Zhang Jingjun, Cheng Liang, Pan Song, Li Yuanhai
Department of Anesthesiology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, P.R. China.
Department of Anesthesiology, The Third People's Hospital of Bengbu, Bengbu, Anhui 233000, P.R. China.
Exp Ther Med. 2019 Mar;17(3):1509-1516. doi: 10.3892/etm.2018.7106. Epub 2018 Dec 17.
Several studies have shown that microRNA-375 (miR-375) is frequently downregulated in several types of human cancer including gastric cancer, colorectal cancer and oral squamous cell carcinoma. However, the role of miR-375 in human esophageal cancer remains unknown. In the current study, the expression level of miR-375 was analyzed in 43 esophageal squamous cell carcinoma (ESCC) tissue and matched adjacent normal tissue samples from patients with ESCC by reverse transcription-quantitative polymerase chain reaction. In addition, the expression level of miR-375 was analyzed in ESCC cell lines (KYSE450 and KYSE150) and the human esophageal epithelial cell line Het-1A by the same method. The expression level of miR-375 was significantly downregulated in ESCC tissue samples and cell lines compared with adjacent normal tissue samples and the human esophageal epithelial cell line, respectively. The effect of miR-375 on ESCC cell proliferation was detected by cell counting kit-8 (CCK-8) and colony formation assays. miR-375 overexpression significantly decreased ESCC cell proliferation and colony formation. Bioinformatics analysis was used to predict specificity protein 1 (SP1) as a target gene of miR-375 in ESCC, and this was verified by dual-luciferase assay. The present study demonstrated that SP1 regulates ESCC cell proliferation and colony formation through direct interaction with miR-375. In addition, the overall survival of patients with ESCC was analyzed using the Kaplan-Meier method and log-rank test. The results indicated that patients with ESCC with high miR-375 expression had a better survival rate compared with patients with ESCC with low miR-375 expression. Taken together, these results suggest that downregulated miR-375 promotes ESCC cell proliferation and colony formation via direct targeting of SP1, and this association may contribute to ESCC progression.
多项研究表明，微小RNA-375（miR-375）在包括胃癌、结直肠癌和口腔鳞状细胞癌在内的多种人类癌症中经常下调。然而，miR-375在人类食管癌中的作用尚不清楚。在本研究中，通过逆转录-定量聚合酶链反应分析了43例食管鳞状细胞癌（ESCC）组织及来自ESCC患者的配对癌旁正常组织样本中miR-375的表达水平。此外，采用相同方法分析了ESCC细胞系（KYSE450和KYSE150）及人食管上皮细胞系Het-1A中miR-375的表达水平。与癌旁正常组织样本及人食管上皮细胞系相比，ESCC组织样本和细胞系中miR-375的表达水平均显著下调。通过细胞计数试剂盒-8（CCK-8）和集落形成试验检测miR-375对ESCC细胞增殖的影响。miR-375过表达显著降低了ESCC细胞的增殖和集落形成。生物信息学分析预测特异性蛋白1（SP1）为ESCC中miR-375的靶基因，并通过双荧光素酶试验进行了验证。本研究表明，SP1通过与miR-375直接相互作用调节ESCC细胞的增殖和集落形成。此外，采用Kaplan-Meier法和对数秩检验分析了ESCC患者的总生存期。结果表明，与miR-375低表达的ESCC患者相比，miR-375高表达的ESCC患者生存率更高。综上所述，这些结果表明，miR-375下调通过直接靶向SP1促进ESCC细胞增殖和集落形成，这种关联可能促进ESCC进展。
