摘要
目的探讨miRNA-29b-3p在食管鳞癌(Esophageal squamous cell carcinoma,ESCC)中的表达水平及其对ESCC细胞生物学行为的影响。方法通过癌症基因组图谱(The cancer genome atlas,TCGA)分析miRNA-29b-3p在恶性肿瘤中的表达,实时荧光定量聚合酶链反应(Quantitative real-time polymerase chain reaction,qRT-PCR)分析15例ESCC癌组织及癌旁正常组织中miRNA-29b-3p的表达情况。将阴性对照抑制剂(inhibitor-NC)和miRNA-29b-3p抑制剂(miRNA-29b-3p-inhibitor)分别转染至TE-1和ECA109细胞,同时以未转染的细胞作为空白对照进行实验。通过细胞增殖实验、流式细胞术、细胞迁移和细胞侵袭实验分析miRNA-29b-3p对TE-1和ECA109细胞增殖、细胞周期、细胞凋亡和细胞迁移及细胞侵袭的影响。结果miRNA-29b-3p在ESCC癌组织中的相对表达量为(0.55±0.20),癌旁组织中的相对表达量为(1.03±0.27),差异有统计学意义(t=5.611,P<0.05)。TCGA数据库分析也显示miRNA-29b-3p在包括食管癌在内的多种恶性肿瘤中低表达。在TE-1和ECA109细胞中成功转染miRNA-29b-3p抑制剂后,细胞增殖活性显著增强(P均<0.05)。流式细胞术检测显示,抑制miRNA-29b-3p可使TE-1细胞G0/G1期和S期细胞比例减少,G2/M期细胞比例增加(P均<0.05);而ECA109细胞中miRNA-29b-3p-inhibitor组的G0/G1期细胞比例较control组与inhibitor-NC组降低,而S期及G2/M期的细胞比例较control组与inhibitor-NC组升高(P<0.05)。沉默miRNA-29b-3p对TE-1和ECA109细胞凋亡的影响差异无统计学意义(P>0.05)。Transwell实验结果表明,沉默miRNA-29b-3p可显著增加TE-1和ECA109细胞的迁移和侵袭能力(P均<0.05)。结论miRNA-29b-3p在ESCC中低表达,可通过调节细胞周期进而调控ESCC细胞的增殖、迁移与侵袭能力。miRNA-29b-3p可成为ESCC诊治的生物标志物。
Objective To investigate the expression level of miRNA-29b-3p in esophageal squamous cell carcinoma(ESCC)and its influence on the biological behaviors of ESCC cells.Methods The expression of miRNA-29b-3p in malignant tumors was analyzed using the cancer genome atlas database,and the expres-sion of miRNA-29b-3p in 15 ESCC tissues and paracancerous tissues was analyzed by quantitative real-time polymerase chain reaction.Negative control inhibitors and miRNA-29b-3p inhibitors were transfected into TE-1 and ECA109 cells respectively,with non-transfected cells serving as blank controls.The effects of miRNA-29b-3p on TE-1 and ECA109 cell proliferation,cell cycle,apoptosis,migration and invasion were analyzed through cell proliferation assays,flow cytometry,and cell migration and invasion assays.Results The relative expression of miRNA-29b-3p in ESCC cancer tissues was(0.55±0.20),and in adjacent tissues was(1.03±0.27).The difference in miRNA-29b-3p expression was statistically significant(t=5.611,P<0.05).TCGA database analysis also showed that miRNA-29b-3p was lowly expressed in various malig-nant tumors including esophageal cancer.After successful transfection of miRNA-29b-3p inhibitor into TE-1 and ECA109 cells,the cell proliferation activity was significantly enhanced(all P<0.05).Flow cy-tometry detection showed that inhibition of miRNA-29b-3p could reduce the proportion of G0/G1 and S phase cells and increase the proportion of G2/M phase cells in TE-1 cells(all P<0.05);while in ECA109 cells,the proportion of G0/G1 phase cells in the miRNA-29b-3p-inhibitor group was lower than that in the control group and the inhibitor-NC group,and the proportion of S and G2/M phase cells was higher than that in the control group and the inhibitor-NC group(P<0.05).Inhibition of miRNA-29b-3p had no sta-tistically significant effect on the apoptosis of TE-1 and ECA109 cells(P>0.05).Transwell assay results indicated that inhibition of miRNA-29b-3p could significantly increase the migration and invasion abilities of TE-1 and ECA109 cells(all P<0.05).Conclusion The low expression of miRNA-29b-3p in ESCC,miRNA-29b-3p may affect the proliferation,migration and invasion ability of ESCC cells by regulating the cell cycle.MiRNA-29b-3p can be used as a biomarker for diagnosis and treatment of ESCC.
作者
范志勤
米尔阿力木江·麦麦提吐尔逊
马遇庆
FAN Zhiqin;Mieralimujiang Maimaitituerxun;MA Yuqing(Department of Pathology,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China;Department of Day Surgery,the Affiliated Tumor Hospital of Xinjiang Medical University,Urumqi 830011,China)
出处
《新疆医科大学学报》
2025年第12期1614-1621,1627,共9页
Journal of Xinjiang Medical University
基金
省部共建中亚高发病成因与防治国家重点实验室开放课题项目(SKL-HIDCA-2021-20)。
