摘要
目的筛选并鉴定与子宫内膜容受性(endometrial receptivity,ER)密切相关的lncRNA:lncAC079944.2,并进一步探究其调节ER的作用与机制。方法根据对GEO数据库的两个数据集共71个子宫内膜组织样本转录组数据的加权基因共表达网络分析(weighted gene co-expression network analysis,WGCNA),筛选出与ER显著相关的RNAs。对2021年10月至2022年10月期间在江苏大学第四附属医院生殖中心就诊患者的子宫内膜样本分析验证生物信息学筛选的RNAs与其临床表达的一致性。应用细胞功能学实验分析筛选的lncAC079944.2对Ishikawa细胞(子宫内膜癌细胞,替代子宫内膜细胞)增殖、迁移和侵袭等功能的影响。通过双荧光素酶报告基因、qRT-PCR、免疫印迹实验等验证lncAC079944.2、EFNA1的表达模式以及和筛选的miRNA之间的相互作用关系,并验证三者在细胞功能上的影响。结果LncAC079944.2在分泌中期表达较分泌早期明显上调(P<0.001),反复种植失败(recurrent implantation failure,RIF)患者分泌中期表达较正常患者分泌中期明显降低(P<0.001)。在干扰lncAC079944.2的表达后,Ishikawa细胞的增殖(P=0.004)、迁移(P=0.001)与侵袭(P<0.001)能力均有不同程度的减弱。qRT-PCR结果显示lncAC079944.2的表达后,EFNA1的表达也呈现下降趋势(P=0.030),而加入miR-149-5p inhibitor后,EFNA1的表达有所恢复(P=0.034)。同样地,在加入miR-149-5p inhibitor后,Ishikawa细胞因lncAC079944.2的表达受到干扰而减弱的增殖(P<0.001)、迁移(P=0.001)与侵袭(P=0.008)能力有所恢复。在胚胎黏附实验中,干扰lncAC079944.2的表达会抑制JAR细胞球体(模拟胚胎)和Ishikawa细胞之间的黏附(P<0.001),而加入miR-149-5p inhibitor后,胚胎和Ishikawa细胞之间的黏附程度回升(P<0.001)。结论LncAC079944.2与ER相关,下调lncAC079944.2可显著降低ER。LncAC079944.2可作为miR-149-5p的分子海绵,调节EFNA1的表达,影响Ishikawa细胞的增殖、迁移与侵袭能力,并影响�
ObjectiveTo screen and identify the lncRNA AC079944.2,which is closely associated with endometrial receptivity(ER),and further delved into its functional role and regulatory mechanism in ER.MethodsWeighted gene co-expression network analysis(WGCNA)was performed on transcriptome data obtained from 71 endometrial tissue samples sourced from two datasets in the GEO database,leading to the identification of RNAs exhibiting significant correlation with ER.The consistency of bioinformatic-screened RNAs and their clinical expression was verified by analysis of endometrial samples from patients treated in the Department of Reproductive Medicine,the Fourth Affiliated Hospital of Jiangsu University from October 2021 to October 2022.Subsequently,the impact of lncAC079944.2 on Ishikawa cells'proliferation,migration,invasion was evaluated through cytofunctional experiments.The expression patterns of lncAC079944.2 and EFNA1,along with their interactions with screened miRNAs,were validated using dual luciferase reporter gene assays,qRT-PCR,and Western blotting analysis.Furthermore,their effects on cell function were confirmed.ResultsLncAC079944.2 was significantly upregulated in the middle secretory stage compared with the early secretory stage(P<0.001),and its expression in the middle secretory stage of recurrent implantation failure(RIF)patients was significantly lower than that of normal patients(P<0.001).Silencing the expression of lncAC079944.2 resulted in varying degrees of weakened proliferation(P=0.004),migration(P=0.001),and invasion(P<0.001)of Ishikawa cells.qRT-PCR revealed that silence of lncAC079944.2 led to a decrease in the expression of EFNA1(P=0.030),whereas the addition of miR-149-5p inhibitor resulted in the recovery of EFNA1 expression(P=0.034).Similarly,the addition of miR-149-5p inhibitor restored the weakened proliferation(P<0.001),migration(P=0.001)and invasion(P=0.008)abilities of Ishikawa cells,which had been compromised by silence of lncAC079944.2.In the embryo adhesion experiment,disruption of lncAC0
作者
商俊宇
陈雨美
朱小兰
Shang Junyu;Chen Yumei;Zhu Xiaolan(The Department of Reproductive Medicine,the Fourth Affiliated Hospital of Jiangsu University,Zhenjiang 212000,China;The Department of Gynecology,the Fourth Affiliated Hospital of Jiangsu University,Zhenjiang 212000,China)
出处
《中华生殖与避孕杂志》
CAS
CSCD
北大核心
2024年第7期705-715,共11页
Chinese Journal of Reproduction and Contraception
基金
国家自然科学基金(82172838)
江苏省自然科学基金(BK20201227)。
