摘要
目的筛选宫颈癌中差异表达的微小RNA(miRNA,miR),探讨关键miRNA对宫颈癌发生发展的作用及其机制。方法采用生物信息学方法分析并以差异倍数>2.5及P<0.01为条件筛选宫颈鳞癌中差异表达的miRNA;采用原位杂交和qRT-PCR分别检测miR-205及白细胞介素(IL)-32的表达;采用CCK-8法检测细胞增殖活力;应用Transwell模型评价细胞迁移及侵袭能力;利用Western blotting检测侵袭相关基质金属蛋白酶(MMP)-2及MMP-9蛋白的表达量。结果生物信息学分析获得宫颈鳞癌中差异表达的miRNA 106个,其中70个上调,36个下调;其中miR-205的表达量变化最大。原位杂交及qRT-PCR结果表明,miR-205在宫颈鳞癌细胞中的表达量明显高于癌旁组织(P<0.05)。miR-205可明显提升宫颈癌细胞的增殖、迁移及侵袭能力(P<0.05)。在宫颈癌细胞系HeLa及SiHa细胞中,过表达miR-205可增强IL-32的表达,降低miR-205表达则可抑制IL-32的表达(P<0.05)。荧光素酶报告基因检测结果显示,miR-205可与IL-32启动子作用提高靶基因的表达量(P<0.05);降低miR-205及IL-32表达均可抑制细胞的侵袭能力(P<0.05);IL-32表达降低能明显抑制侵袭相关蛋白MMP-2及MMP-9的表达(P<0.05)。结论宫颈鳞癌中miR-205表达量明显增高,可能通过上调IL-32的表达促进MMP-2及MMP-9的表达,最终促进宫颈癌细胞的增殖、迁移及侵袭。
Objective To identify the differentially expressed microRNA(miRNA,miR)in cervical squamous cell carcinoma(SCC)and explore the mechanism of critical miRNA on the occurrence and development of cervical cancer.Methods To identify differentially expressed miRNA,we employed bioinformatics methods to filter out miRNA under the conditions of|fold change(FC)|>2.5 and P<0.01.The RNA expressions of miR-205 and interleukin-32(IL-32)were detected by in situ hybridization and qRT-PCR,respectively.We evaluated cell proliferation,migration,and invasion through CCK-8 and Transwell model.The protein level of matrix metalloproteinase-3(MMP-2)and MMP-9 were quantified by Western blotting.Results Through bioinformatics analysis,we obtained 106 differentially expressed miRNA in cervical cancer,with 70 up-regulated and 36 down-regulated miRNAs.miR-205 was the most differentially expressed miRNA.We validated the overexpression of miR-205 in cervical squamous cell carcinoma by in situ hybridization and qRT-PCR(P<0.05).Overexpression of miR-205 could significantly promote the proliferation,migration,and invasion of cervical cancer cells(P<0.05).The overexpression or decreased expression of miR-205 can increase or inhibit the expression of IL-32 in cervical cancer cell lines HeLa and SiHa,and miR-205 can directly interact with the IL-32 promoter to increase the expression of target genes(P<0.05);the decreased expression of miR-205 and IL-32 inhibited the invasion of cells(P<0.05);moreover,the decreased expression of IL-32 could significantly inhibit the expression of the invasion-associated proteins MMP-2 and MMP-9(P<0.05).Conclusion In cervical squamous cell carcinoma,miR-205 is dramatically up-regulated,potentially promoting the proliferation,migration,and invasion of cancer cells,which could further enhance the expression of MMP-2 and MMP-9 through IL-32 up-regulation,and ultimately improve the invasion ability of cervical cancer cells.
作者
刘建兵
林晓雨
李文龙
王伟
王文豪
崔小华
郝建卿
李莉
郝敏
Liu Jian-Bing;Lin Xiao-Yu;Li Wen-Long;Wang Wei;Wang Wen-Hao;Cui Xiao-Hua;Hao Jian-Qing;Li Li;Hao Min(Departments of Obstetrics and Gynecology,the Second Hospital of Shanxi Medical University,Taiyuan,Shanxi 030001,China;Department of Medical Cell Biology and Genetics,School of Basic Medical,Shanxi Medical University,Taiyuan,Shanxi 030001,China)
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2022年第7期694-700,共7页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金(81972452)
山西省基础研究计划(20210302123320)
山西省重点研发计划(201803D31121,201903D321152)
国家卫生和计划生育委员会公益性行业科研专项(201402010)。
关键词
宫颈肿瘤
癌
鳞状细胞
miR-205
细胞增殖
细胞侵袭
细胞运动
uterine cervical neoplasms
carcinoma,squamous cell
miR-205
cell proliferation
cell invasion
cell movement
