miR-107对宫颈癌细胞系HeLa生物学行为的影响  

Influence of miR-107 on biological behaviors of HeLa cell line

作　　者：刘雅坤[1] 杜家宜 赵运清 LIU Ya-kun;DU Jia-yi;ZHAO Yun-qing(Department of Gynecology,the Fourth Hospital of Hebei Medical University,Shijiazhuang,Hebei 050000,China;Department of Radiotherapy and Chemotherapy,the Fourth Hospital of Hebei Medical University,Shijiazhuang,Hebei 050000,China;Department of Gynecology,Civil Affairs General Hospital of Hebei Province,Xingtai,Hebei 054000,China)

机构地区：[1]河北医科大学第四医院妇科,河北石家庄050000 [2]河北医科大学第四医院放化疗科,河北石家庄050000 [3]河北省民政总医院妇科,河北邢台054000

出　　处：《中华实用诊断与治疗杂志》2021年第10期1001-1006,共6页Journal of Chinese Practical Diagnosis and Therapy

基　　金：2020年度河北省医学科学研究课题计划(20201088)。

摘　　要：目的探讨miR-107调控氧连接氮乙酰葡萄糖胺转移酶(O-linked N-acetylglucosamine transferase, OGT)对宫颈癌细胞HeLa增殖、迁移和侵袭的影响。方法 40例宫颈癌患者均行手术治疗,取其手术切除的宫颈癌组织和癌旁组织,采用实时荧光定量PCR法检测2种组织miR-107和OGT mRNA相对表达量。对数生长期宫颈癌细胞HeLa随机分为对照组、模拟剂对照组、模拟剂组、抑制剂对照组、抑制剂组,对照组细胞正常培养,模拟剂对照组、模拟剂组、抑制剂对照组、抑制剂组细胞分别转染mimic-NC、miR-107 mimic、inhibitor-NC、miR-107 inhibitor,转染48 h后,采用MTT法检测细胞存活率,采用划痕实验检测细胞迁移距离,采用Transwell法检测侵袭细胞率,采用Western blot法检测细胞OGT、上皮钙黏蛋白、神经钙黏蛋白相对表达量,采用双荧光素酶报告基因法检测miR-107与OGT的靶向关系。结果宫颈癌组织miR-107相对表达量(3.53±0.17)低于癌旁组织(14.82±0.22)(P<0.05),OGT mRNA相对表达量(7.49±0.25)高于癌旁组织(5.38±0.24)(P<0.05);模拟剂组细胞存活率[(47.53±3.36)%]、划痕愈合百分比[(12.62±2.11)%]低于对照组[(99.93±1.02)%、(26.40±2.16)%]、模拟剂对照组[(99.85±2.11)%、(25.73±2.24)%]、抑制剂组[(99.96±3.24)%、(75.33±3.50)%](P<0.05),侵袭细胞数[(23.40±5.16)个]少于对照组[(82.83±6.47)个]、模拟剂对照组[(79.61±6.28)个]、抑制剂组[(121.44±5.10)个](P<0.05);抑制剂组细胞划痕愈合百分比高于对照组、抑制剂对照组[(26.48±3.02)%](P<0.05),侵袭细胞数多于对照组、抑制剂对照组[(80.93±5.08)个](P<0.05),细胞存活率与抑制剂对照组[(99.93±2.16)%]、对照组比较差异均无统计学意义(P>0.05)。模拟剂组细胞OGT、神经钙黏蛋白相对表达量低于对照组、模拟剂对照组、抑制剂组(P<0.05),上皮钙黏蛋白相对表达量高于对照组、模拟剂对照组、抑制剂组(P<0.05);抑制剂组细胞OGT、神经钙黏蛋白�Objective To investigate the influence of miR-107 on the proliferation, migration and invasion of HeLa cells by regulating O-linked N-acetylglucosamine transferase(OGT). Methods The relative expressions of miR-107 and OGT mRNA were detected by real-time fluorescence quantitative PCR in the resected cervical cancer tissues and paracancerous tissues from 40 patients with cervical cancer. The HeLa cells in logarithmic growth phase were randomly divided into control group, mimic control group, mimic group, inhibitor control group and inhibitor group. The cells in control group were cultured normally. Mimic control group, mimic group, inhibitor control group and inhibitor group were transfected with mimic-NC, miR-107 mimic, inhibitor-NC and miR-107 inhibitor, respectively. After 48-h transfection, MTT assay was used to detect cell survival rate, scratch assay was used to detect cell migration distance, Transwell assay was used to detect cell invasion rate, and Western blot was used to detect the relative expressions of OGT, E-cadherin and N-cadherinproteins.Dual luciferase reporter assay was used to detect the targeting relationship between miR-107 and OGT.Results The relative expression of miR-107 was lower in cervical cancer tissues(3.53±0.17)than that in paracancerous tissues(14.82±0.22)(P<0.05),and the relative expression of OGT mRNA was higher in cervical cancer tissues(7.49±0.25)than that in paracancerous tissues(5.38±0.24)(P<0.05).The cell survival rate and the percentage of scratch healing were lower in mimic group [(47.53±3.36)%,(12.62±2.11)%]than those in control group [(99.93±1.02)%,(26.40±2.16)%],mimic control group [(99.85±2.11)%,(25.73±2.24)%]and inhibitor group[(99.96±3.24)%,(75.33±3.50)%](P<0.05),and the number of invasive cells was less in mimic group(23.40±5.16)than that in control group(82.83±6.47),mimic control group(79.61±6.28)and inhibitor group(121.44±5.10)(P<0.05).The percentage of cell scratch healing was higher in inhibitor group than that in control group and inhibitor contro

关 键 词：宫颈癌 miR-107 氧连接氮乙酰葡萄糖胺转移酶 上皮钙黏蛋白 神经钙黏蛋白 

分 类 号：R737.33[医药卫生—肿瘤]