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浙江大学学报（医学版）
浙江大学学报（医学版）  2016, Vol. 45 Issue (4): 342-348    DOI: 10.3785/j.issn.1008-9292.2016.07.03
癌分子医学专题     
微RNA-let-7e-3p在宫颈上皮内瘤变和宫颈癌组织中的表达及临床意义
陈晓静, 许君芬, 叶菁, 程晓东, 谢幸, 吕卫国
浙江大学医学院附属妇产科医院中心实验室 浙江省女性生殖健康研究重点实验室, 浙江 杭州 310006
Expression of miR-let-7e-3p in cervical intraepithelial neoplasm and cervix carcinoma and its clinical significance
CHEN Xiaojing, XU Junfen, YE Jing, CHENG Xiaodong, XIE Xing, LYU Weiguo
Central Laboratory, Women's Hospital, Zhejiang University School of Medicine, Key Laboratory of Women's Reproductive Health of Zhejiang Province, Hangzhou 310006, China
全文: PDF(1118 KB)  
摘要: 

目的：探索微RNA（miRNA，miR-）-let-7e-3p在不同宫颈病变中表达情况及临床意义。方法：收集并整理具有完整临床病理资料的病例，其中正常子宫颈26例、子宫颈高级别鳞状上皮内病变（HSIL）37例、宫颈癌101例；采用实时定量RT-PCR检测临床宫颈组织样本中miR-let-7e-3p的相对表达量，分析miR-let-7e-3p表达水平与宫颈癌的发生及临床不良预后因素间的相关性。将miR-let-7e-3p模拟物转染至宫颈癌细胞株Siha（转染组），采用流式细胞仪检测Siha细胞周期和细胞凋亡变化；CCK-8试剂盒检测Siha细胞增殖变化；Transwell实验检测Siha细胞迁移和侵袭能力变化。结果：miR-let-7e-3p在正常宫颈、HSIL及宫颈癌组织中的相对表达量分别为1.45±0.24、0.79±0.05、0.46±0.04，正常子宫颈与HSIL、正常子宫颈与宫颈癌、HSIL与宫颈癌相对表达量差异均具有统计学意义（均P<0.05）；miR-let-7e-3p表达水平与患者宫颈癌临床不良预后因素之间无显著相关性；转染组S期细胞（29.76%±6.60%）高于阴性对照组（13.38%±1.30%，P<0.05）；转染组凋亡率（5.98%±1.38%）高于阴性对照组（3.53%±0.79%，P<0.05）；转染组转染后48、72、96 h吸光度值分别为0.57±0.11、0.65±0.04、0.84±0.14，阴性对照组为0.74±0.05、0.93±0.10、1.47±0.14，细胞增殖均被抑制（均P<0.05）；转染组与阴性对照组迁移和侵袭能力差异均无统计学意义（均P>0.05）。结论：miR-let-7e-3p可能在宫颈癌的发生中发挥抑癌基因的作用，阻滞细胞周期和抑制细胞增殖可能是其重要调控机制。
关键词 ： 微RNAs,  宫颈肿瘤/病理学,  宫颈上皮内瘤样病变/病理学,  基因,肿瘤抑制,  细胞周期,  细胞增殖,  基因表达,  逆转录聚合酶链反应    
Abstract：

Objective: To investigate the expression of microRNA (miRNA, miR) let-7e-3p in different cervical lesions and its clinical significance. Methods: The expression of miR-let-7e-3p in the tissues of normal cervix (n=26), high-grade squamous intraepithelial lesion (HSIL) (n=37), and cervix carcinoma (n=101) were detected by reverse transcription and quantitative polymerase chain reaction (RT-qPCR). The correlation of miR-let-7e-3p expression with the clinicopathological parameters of patients with cervical cancer was analyzed. miR-let-7e-3p mimic was transfected into cervical carcinoma Siha cells. The cell cycle and apoptosis were determined by flow cytometry; cell proliferation was determined by CCK-8 kit; and the migration and invasion of cells were determined by Transwell assay. Results: The relative expression levels of miR-let-7e-3p in normal cervix, HSIL, and cervical carcinoma were 1.45±0.24, 0.79±0.05 and 0.46±0.04, respectively (all P<0.05). After transfection with miR-let-7e-3p mimic, the S-phase fraction and apoptosis rate of Siha cells were increased significantly compared with control group[(29.76±6.6)% vs (13.38±1.3)%, P<0.05; (5.98±1.38)% vs (3.53±0.79)%, P<0.05, respectively]. OD of transfected Siha cells at 48, 72 and 96 h were 0.57±0.11,0.65±0.04 and 0.84±0.14, which were significantly lower than those of untransfected Siha cells (0.74±0.05, 0.93±0.10 and 1.47±0.14, all P<0.05). The migration and invasion abilities of transfected Siha cells were not significantly changed (all P>0.05). Conclusion: The expression of miR-let-7e-3p is down-regulated in cervical neoplasms, which is associated with cell cycle arrest and proliferation inhibition of cervical cancer cells.
Key words： MicroRNAs    Uterine cervical neoplasms/pathology    Cervical intraepithelial neoplasia/pathology    Genes, tumor suppressor    Cell cycle    Cell proliferation    Gene expression    Reverse transcriptase polymerase chain reaction
收稿日期: 2015-08-11
CLC:  R737.33  
基金资助:

国家自然科学基金（81472421，81272861）；浙江省自然科学基金（LQ15H160003）；国家重点研发计划“重大慢性非传染性疾病防控研究”重点专项（2016YFC1302900，2016YFC0902900）
通讯作者: 吕卫国(1965-),博士,主任医师,教授,博士生导师,主要从事妇科肿瘤临床和基础研究;E-mail:lbwg@zju.edu.cn;http://orcid.org/0000-0002-8570-7317     E-mail: lbwg@zju.edu.cn
作者简介: 陈晓静(1986-),学士,技师,主要从事妇科肿瘤基础研究;E-mail:rj045wwq@163.com;http://orcid.org/0000-0001-8107-9037
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引用本文:

陈晓静 等. 微RNA-let-7e-3p在宫颈上皮内瘤变和宫颈癌组织中的表达及临床意义[J]. 浙江大学学报（医学版）, 2016, 45(4): 342-348.
CHEN Xiaojing, XU Junfen, YE Jing, CHENG Xiaodong, XIE Xing, LYU Weiguo. Expression of miR-let-7e-3p in cervical intraepithelial neoplasm and cervix carcinoma and its clinical significance. Journal of ZheJiang University(Medical Science), 2016, 45(4): 342-348.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2016.07.03      或      http://www.zjujournals.com/xueshu/med/CN/Y2016/V45/I4/342

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