Please wait a minute...
Journal of ZheJiang University(Medical Science)  2016, Vol. 45 Issue (4): 342-348    DOI: 10.3785/j.issn.1008-9292.2016.07.03
    
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
Download:   PDF(1118KB)
Export: BibTeX | EndNote (RIS)      

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 wordsMicroRNAs      Uterine cervical neoplasms/pathology      Cervical intraepithelial neoplasia/pathology      Genes, tumor suppressor      Cell cycle      Cell proliferation      Gene expression      Reverse transcriptase polymerase chain reaction     
Received: 11 August 2015     
CLC:  R737.33  
  R730.23  
Cite this article:

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.

URL:

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


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

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 
[[1]]   TORRE L A, BRAY F, SIEGEL R L, et al. Global cancer statistics, 2012[J]. CA Cancer J Clin,2015,65(2):87-108.
[[2]]   ARBYN M, CASTELLSAGUE X, DE S S, et al. Worldwide burden of cervical cancer in 2008[J]. Ann Oncol,2011,22(12):2675-2686.
[[3]]   LI Y,WANG F F, XU J F, et al. Progressive miRNA expression profiles in cervical carcinogenesis and identification of HPV-related target genes for miR-29[J]. J Pathol,2011, 224(4):484-495.
[[4]]   XU J F, LI Y, WANG F F, et al. Suppressed miR-424 expression via upregulation of target gene Chk1 contributes to the progression of invasive cervical cancer[J]. Oncogene,2013,32(8):976-987.
[[5]]   VILLEGAS R V, JUAREZ M S, PEREZ G O, et al. Heterogeneity of microRNAs expression in invasive cervical cancer cells: over-expression of miR-196a[J]. Int J Clin Exp Pathol,2014,7(4):1389-1401.
[[6]]   ROUSH S, SLACK F J. The let-7 family of microRNAs[J]. Trends Cell Biol, 2008,18(10):505-516.
[[7]]   BOYERINAS B, PARK S M, HAU A, et al. The role of let-7 in cell differentiation and cancer[J]. Endocr Relat Cancer,2010,17(1):F19-36.
[[8]]   CHIU S C, CHUNG H Y, CHO D Y, et al. Therapeutic potential of microRNA let-7: tumor suppression or impeding normal stemness[J]. Cell Transplant,2014,23(4-5):459-469.
[[9]]   TAKAMIZAWA J, KONISHI H, YANAGISAWA K, et al. Reduced expression of the let-7 microRNAs in human lung cancers in association with shortened postoperative survival[J]. Cancer Res, 2004,64(11):3753-3756.
[[10]]   YANAIHARA N, CAPLEN N, BOWMAN E, et al. Unique microRNA molecular profiles in lung cancer diagnosis and prognosis[J]. Cancer Cell, 2006,9(3):189-198.
[[11]]   SCHMITTGEN T D, LIVAK K J. Analyzing real-time PCR data by the comparative CT method[J]. Nat Protoc, 2008, 3(6):1101-1108.
[[12]]   KONG Y W, FERLAND-MCCOLLOUGH D, JACKSON T J, et al. microRNAs in cancer management[J]. Lancet Oncol, 2012, 13(6):e249-258.
[[13]]   MO Y Y. MicroRNA regulatory networks and human disease[J]. Cell Mol Life Sci, 2012, 69(21):3529-3531.
[[14]]   REINHART B J, SLACK F J, BASSON M, et al. The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans[J]. Nature,2000,403(6772):901-906.
[[15]]   JOHNSON C, ESQUELA K A, STEFANI G, et al. The let-7 microRNA represses cell proliferation pathways in human cells[J]. Cancer Res,2007,67(16):7713-7722.
[[16]]   CHANG C J, HSU C C, CHANG C H, et al. Let-7d functions as novel regulator of epithelial-mesenchymal transition and chemoresistant property in oral cancer[J]. Oncol Rep,2011,26(4):1003-1010.
[[17]]   WANGNER S, NGEZAHAYO A, MURUA E H, et al. Role of miRNA let-7 and its major targets in prostate cancer[J]. Biomed Res Int,2014,2014: 376326.
[[18]]   ZHAO B, HAN H, CHEN J, et al. MicroRNA let-7c inhibits migration and invasion of human non-small cell lung cancer by targeting ITGB3 and MAP4K3[J]. Cancer Lett,2014,342(1):43-51.
[[19]]   HUR K, TOIYAMA Y, SCHETTER A J, et al. Identification of a metastasis-specific MicroRNA signature in human colorectal cancer[J]. J Nat Cancer Inst,2015,107(3):dju492.
[1] YANG Xiaohong, YANG Kun, LIAO Li, JIN Yan. Effect of miR-705 on osteogenic differentiation of mouse embryo osteoblast precursor cells MC3T3-E1[J]. Journal of ZheJiang University(Medical Science), 2016, 45(6): 575-580.
[2] HOU Shifang, WANG Zhihua, WANG Jun, HE Zhixu, SHU Liping. Myeloid and erythroid hematopoietic transcription factor expression decline after knockdown of lmna genes in zebrafish embryos[J]. Journal of ZheJiang University(Medical Science), 2016, 45(6): 620-625.
[3] CAO Peng, LENG Dongjin, LI Ying, ZHANG Ziwei, LIU Lei, LI Xiaoyan. Progress on anti-tumor molecular mechanisms of dihydroartemisinin[J]. Journal of ZheJiang University(Medical Science), 2016, 45(5): 501-507.
[4] WU Zhihua, JING Min, LIANG Hanying, YANG Rong, HUANG Yaping, CHEN Xiaoming, HU Jianhua, FAN Jun. T cell receptor β-chain CDR3 spectratyping and cytomegalovirus activation in allogeneic hematopoietic stem cell transplant recipients[J]. Journal of ZheJiang University(Medical Science), 2016, 45(5): 515-521.
[5] YANG Suwen, WANG Wei, JIN Hong, ZHONG Yuhong, XIE Xinyou. Expression of microRNA-221/222 in patients with monoclonal gammopathy of undetermined significance and multiple myeloma[J]. Journal of ZheJiang University(Medical Science), 2016, 45(4): 371-378.
[6] LOU Pengrong, SUN Xiaonan, ZHOU Jundong, ZOU Shitao. Effect of RAD18-siRNA on proliferation and chemotherapy sensitivity of human esophageal squamous cell carcinoma ECA-109 cells[J]. Journal of ZheJiang University(Medical Science), 2016, 45(4): 364-370.
[7] LIN Weiren, CHEN Yatian, ZENG Linghui, YING Rongbiao, ZHU Feng. Effect of a novel EZH2 inhibitor GSK126 on prostate cancer cells[J]. Journal of ZheJiang University(Medical Science), 2016, 45(4): 356-363.
[8] WANG Cheng, WANG Wenjun, YANG Wei, YU Xiaohua, YAN Yiguo, ZHANG Jian, JIANG Zhisheng. MicroRNAs: a type of novel regulative factor for intervertebral disc degeneration[J]. Journal of ZheJiang University(Medical Science), 2016, 45(2): 170-178.
[9] YANG Wan-hua, WU Hai-ying, ZHANG Hong-ze, LIU Hong-xiang, WEI Yu-jie, SHI Bin. Prognostic value of Picco monitoring combined with plasma microRNA-150 detection in septic shock patients[J]. Journal of ZheJiang University(Medical Science), 2015, 44(6): 659-664.
[10] YANG Yan, LI Yu-mei, ZHANG Na, LI Wan-yun, OU Yu-rong, WANG Rui, ZHAO Fu-you, WU Qiong. Expression of gap junction protein connexin 26 in human hepatocellular carcinoma and its significance[J]. Journal of ZheJiang University(Medical Science), 2015, 44(5): 517-524.
[11] ZHAO Zun-lan, CAI Ying, WANG Yang-yang, XIA Chun-lei, LI Cong-xin, CHEN Su-lian, YANG Qing-ling, CHEN Chang-jie. Effects of miRNA-21 on paclitaxel-resistance in human breast cancer cells[J]. Journal of ZheJiang University(Medical Science), 2015, 44(4): 400-409.
[12] XIN Liao-bing, JIANG Xiu-xiu, YE Xiao-lei, WU Rui-jin, XU Kai-hong, MA Jun-yan, LIN Jun. AQP5 gene silencing inhibits proliferation and migration of ectopic endometrial glandular epithelial cells in endometriosis[J]. Journal of ZheJiang University(Medical Science), 2015, 44(3): 285-292.
[13] WANG Xiao-jun, ZHANG Hao, ZHAN Hong-sheng, DING Dao-fang. Establishment of chondrocyte degeneration model in vitro by rat serum[J]. Journal of ZheJiang University(Medical Science), 2015, 44(3): 308-314.
[14] HAN Yan-xia, YOU Liang-shun, LIU Hui, MAO Li-ping, YE Xiu-jin, QIAN Wen-bin. Apoptosis of acute myeloid leukemia HL-60 cells induced by CDK inhibitor SNS-032 and its molecular mechanisms[J]. Journal of ZheJiang University(Medical Science), 2015, 44(2): 174-178.
[15] ZHU Hai-peng, YUN Feng, JIU Tao, et al. Inhibitory effect of inositol hexaphosphate on proliferation of LNCaP cells and its  relation to IGFBP-3 expression[J]. Journal of ZheJiang University(Medical Science), 2014, 43(5): 521-.