长链非编码RNA-AP001528.2通过靶向调控miR-149-5p介导宫颈癌的发生发展  

Long non-coding RNA-AP001528.2 mediates cervical carcinogenesis through targeted regulation of miR-149-5p

作　　者：叶文蔚[1] 应柳青 蒋梦亭 修丽梦 柴芝红 YE Wenwei;YING Liuqing;JIANG Mengting(Department of Gynecology,Taizhou Municipal Hospital,Taizhou 318000,China)

机构地区：[1]浙江省台州市立医院妇科,浙江台州318000

出　　处：《全科医学临床与教育》2025年第4期298-302,F0002,F0003,共7页Clinical Education of General Practice

基　　金：浙江省医药卫生科技计划项目(2022KY1393);台州市科技计划项目(20ywa37)。

摘　　要：目的研究长链非编码RNA AP001528.2对宫颈癌C-33A细胞增殖、迁移和侵袭能力的影响,并探讨其调控机制。方法采用qRT-PCR实验检测61例宫颈癌组织及癌旁组织中miR-149-5p的表达量,再分析AP001528.2和miR-149-5p的表达水平的相关性;采用CCK-8法、Transwell测定与划痕实验方法,检测AP001528.2基因表达下调对C-33A细胞增殖活性、迁移速率及侵袭潜能的影响变化;荧光原位杂交试验检测miR-149-5p与AP001528.2在宫颈癌组织和细胞中的共定位情况,通过生物信息学预测网站预测到miR-149-5p与AP001528.2有互补结合位点,并利用双荧光素酶报告系统鉴定它们的靶向关系;再记录敲低或者上调AP001528.2表达后miR-149-5p的表达变化情况;将miR-149-5p inhibitor与siRNA-AP001528.2共转染C-33A细胞后,检测C-33A细胞增殖、迁移和侵袭情况。结果在宫颈癌组织中miR-149-5p呈低表达,且与AP001528.2的表达呈负相关;敲低AP001528.2的表达可以明显抑制C-33A细胞的增殖、迁移和侵袭能力;AP001528.2靶向调控miR-149-5p的表达,下调AP001528.2水平能升高miR-149-5p的表达,而上调AP001528.2水平能降低miR-149-5p的表达;下调miR-149-5p表达能明显逆转敲低AP001528.2水平对C-33A细胞增殖、迁移和侵袭能力的抑制作用。AP001528.2与miR-149-5p在细胞质中存在共定位,且两者间存在靶向调控关系。结论AP001528.2与宫颈癌的发展及预后密切相关,AP001528.2可通过靶向调控miR-149-5p表达促进宫颈癌C-33A细胞增殖、迁移和侵袭。Objective To explore the effects of long non-coding RNA AP001528.2 on the growth,motility,and invasive capacity of cervical cancer C-33A cells,and to decipher its fundamental regulatory pathways.Methods The qRT PCR technique was employed to quantify the miR-149-5p expression levels in 61 samples of cervical cancer tissues and their adjacent non-tumor tissues.The relationship between AP001528.2 and miR-149-5p expression levels was analyzed.The proliferation,migration,and invasion abilities of C-33A cells were evaluated using the CCK-8 assay,Transwell migration assay,and wound healing assay after the knockdown of AP001528.2 expression.Fluorescence in situ hybridization(FISH)was used to visualize the spatial coexistence of miR-149-5p and AP001528.2 within cervical cancer tissues and cellular structures.Bioinformatic prediction tools were utilized to identify potential complementary binding sites between miR-149-5p and AP001528.2,and the targeting relationship was further validated by using a dual-luciferase reporter system.The expression of miR-149-5p was quantified by qRT-PCR following the knockdown or upregulation of AP001528.2.Finally,the proliferation,migration,and invasion of C-33A cells were assessed following co-transfection with a miR-149-5p inhibitor and siRNA targeting AP001528.2.Results miR-149-5p was significantly underexpressed in cervical cancer tissues and negatively correlated with the expression of AP001528.2.Knocking down the expression of AP001528.2 significantly inhibited the proliferation,migration,and invasive abilities of C-33A cells.AP001528.2 targeted the regulation of miR-149-5p expression,resulting in a significant decrease in the expression of miR-149-5p.The downregulation of AP001528.2 led to an elevation in miR-149-5p expression,whereas its upregulation reduced miR-149-5p expression.Furthermore,the downregulation of miR-149-5p expression significantly reversed the inhibitory effects of knocking down AP001528.2 on the proliferation,migration,and invasion abilities of C-33A cells.AP001528

关 键 词：AP001528.2 miR-149-5p 宫颈癌 细胞增殖 细胞迁移 细胞侵袭 

分 类 号：R737.33[医药卫生—肿瘤]