[关键词]
[摘要]
目的:探讨 miR-627-3p 在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)组织中的表达及其对 ESCC细胞生物学行为的影响。方法:收集2015年1月至2015年10月河北医科大学第四医院胸外科手术切除的ESCC组织86例及对应的癌旁组织标本20例。通过qPCR法检测miR-627-3p在86例ESCC组织及癌旁组织中的表达,分析其表达与ESCC患者临床病理学指标及预后的关系;利用Kaplan-Meier Plotter在线数据库进一步分析数据库中hsa-miR-627的表达与ESCC患者预后的关系;通过qPCR法检测miR-627-3p在4株ESCC细胞系中的表达,选取表达水平最低的食管癌细胞转染miR-627-3p mimic,选取表达水平最高的ESCC细胞转染miR-627-3p inhibitor,采用CCK-8法检测细胞的增殖,采用Transwell实验检测细胞迁移和侵袭;采用 KEGG分析探讨 miR-627-3p可能介导的信号转导通路,并采用 qPCR法验证 miR-627-3p对信号通路中关键基因表达的影响。结果:miR-627-3p在 ESCC组织中的表达明显低于在癌旁组织中的表达,miR-627-3p的表达与 ESCC患者淋巴结转移和临床分期相关(均 P<0.05);miR-627-3p高表达的 ESCC 患者的 5年生存率明显高于 miR-627-3p低表达的食管癌患者(P<0.05)。ESCC细胞系 KYSE170中 miR-627-3p表达最低,KYSE30中 miR-627-3p表达最高;在 KYSE170细胞中转染 miR-627-3p mimic后,细胞的增殖能力无明显变化(P>0.05),但细胞的迁移(P<0.05)和侵袭能力(P<0.05)显著降低;在 KYSE30 细胞中转染 miR-627-3p inhibitor 后,细胞的增殖能力无明显变化(P>0.05),但细胞的迁移(P<0.05)和侵袭能力(P<0.05)显著增高。KEGG 分析结果显示,miR-627-3p介导了多条与肿瘤相关的信号转导通路。结论:miR-627-3p在ESCC组织中的表达明显低于在癌旁组织中的表达,其低表达与ESCC患者的不良预后相关,miR-627-3p抑制ESCC细胞的迁移和侵袭,可能是通过干扰多条与肿瘤相关的信号通路发挥生物学功能。
[Key word]
[Abstract]
Objective: To investigate the expression of miR-627-3p in esophageal squamous cell carcinoma (ESCC) tissues and its effect on biological behaviours of ESCC cells. Methods: From January 2015 to October 2015, 86 ESCC tissue specimens and 20 cooresponding adjacent normal tissue specimens from Thoracic Surgery Department of the Fourth Hospital of Hebei Medical University were collected. qPCR was used to detect the expression level of miR-627-3p in 86 cases of ESCC tissues and 20 cases of adjacent normal tissues. The relationship between miR-627-3p expression and clinicopathological parameters as well as prognosis was analyzed. The Kaplan-Meier plotter online database was used to further analyze the relationship between hsa-miR-627 expression and the prognosis of ESCC patients. qPCR was used to detect the expression of miR-627-3p in the four ESCC cells. miR-627-3p mimic was transfected into low-expressed cells, and miR-627-3p inhibitor was transfected into high-expressed cells. The cell proliferation was detected by CCK-8 assay, and the cell migration and invasion were detected by Transwell assay, respectively. KEGG analysis was used to explore the possible signal pathways mediated by miR-627-3p, and qPCR was used to verify the effect of miR-627-3p on the expression of key genes in the siganl pathway. Results: The expression of miR-627-3p in ESCC tissues was significantly lower than that in adjacent normal tissues. The expression of miR-627-3p was negatively correlated with lymph node metastasis and clinical stage of ESCC patients (all P<0.05). The five-year survival rate of ESCC patients with high miR-627-3p expression was significantly higher than that of the patients with low miR-627-3p expression (P<0.05). The expression of miR-627-3p was lowest in KYSE170 cells, and highest in KYSE30 cells. After transfection of miR-627-3p mimic into KYSE170 cells, the cell proliferation ability was not changed (P>0.05), but cell migration (P<0.05) and invasion (P<0.05) abilities were significantly decreased. After transfection of miR-627-3p inhibitor into KYSE30 cells, the cell proliferation ability was not changed (P>0.05), but cell migration (P<0.05) and invasion (P<0.05)abilities were significantly increased. KEGG analysis showed that miR-627-3p mediated multiple tumor-related signal pathways.Conclusion: The expression of miR-627-3p in ESCC tissues is significantly lower than in adjacent normal tissues, and its low expression is related to the poor prognosis of ESCC patients. miR-627-3p inhibits the migration and invasion of ESCC cells, which might play a biological function by interfering with multiple tumor-related signal pathways.
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[基金项目]
河北省医学科学研究课题计划资助项目(No.20210074)