Wang Liming, Xing Qi, Feng Tongfu, He Ming, Yu Weixu, Chen Hui
Department of Gynecology, Hubei Provincial Maternal and Child Health Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Gynecology and Obstetrics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
J Cell Biochem. 2020 Feb;121(2):1703-1715. doi: 10.1002/jcb.29406. Epub 2019 Oct 8.
MiR-145 has been shown to suppress cell invasiveness and proliferation in endometriosis, whereas prostate cancer-associated transcript 1 (PCAT1) was reported to act as a sponge of miR-145 with one single-nucleotide polymorphism (SNP), rs710886, located in the chromosomal segment of PCAT1. Therefore, this study aimed to explore the association between rs710886 SNP and the risk of endometriosis, as well as the effect of this SNP on the activation of the signaling pathway downstream of PCAT1. Real-time polymerase chain reaction (PCR) was performed to observe the expression of miR-145 in transfected cells, while Matrigel invasion chamber assays and MTT assay were conducted to examine the invasiveness/proliferation among different cell groups. Moreover, bioinformatics tools, luciferase assays, real-time PCR, and Western blot analysis were used to measure the expression of these target genes in the presence of miR-145. Finally, a statistical analysis was conducted to compare the genotypes of rs710886 SNP between fertile healthy women and infertile women with endometriosis. PCAT1 small interfering RNA (siRNA) evidently increased the expression of miR-145 but reduced the invasiveness/proliferation of cells. P-PCAT1 exhibited an opposite effect as that of PCAT1 siRNA, indicating PCAT1 could promote the proliferation and invasiveness of endometriosis stem cells via inhibiting the expression of miR-145. Meanwhile, FASCIN1, SOX2, MSI2, SERPINE1, and JAM-A were identified as target genes of miR-145 via computational analysis and luciferase assays. Finally, a significant genetic effect was observed in both the dominant (AG+GG vs AA) and recessive models (GG vs AG+AA), indicating the presence of an association between the genotype of SNP rs710886 and the risk of endometriosis. SNP rs710886 A>G could lower the expression of PCAT1, thus leading to the overexpression of miR-145. Highly expressed miR-145 would inhibit the invasiveness and proliferation of endometriosis stem cells via targeting specific genes, thus decreasing the risk of endometriosis.
已有研究表明，miR - 145可抑制子宫内膜异位症中的细胞侵袭和增殖，而前列腺癌相关转录本1（PCAT1）据报道可作为miR - 145的海绵，其位于PCAT1染色体片段上的一个单核苷酸多态性（SNP），即rs710886。因此，本研究旨在探讨rs710886 SNP与子宫内膜异位症风险之间的关联，以及该SNP对PCAT1下游信号通路激活的影响。采用实时聚合酶链反应（PCR）观察转染细胞中miR - 145的表达，同时进行基质胶侵袭小室试验和MTT试验以检测不同细胞组之间的侵袭性/增殖能力。此外，利用生物信息学工具、荧光素酶试验、实时PCR和蛋白质免疫印迹分析来检测在miR - 145存在的情况下这些靶基因的表达。最后，进行统计分析以比较生育健康女性和患有子宫内膜异位症的不孕女性之间rs710886 SNP的基因型。PCAT1小干扰RNA（siRNA）明显增加了miR - 145的表达，但降低了细胞的侵袭性/增殖能力。P - PCAT1表现出与PCAT1 siRNA相反的作用，表明PCAT1可通过抑制miR - 145的表达来促进子宫内膜异位症干细胞的增殖和侵袭。同时，通过计算分析和荧光素酶试验确定FASCIN1、SOX2、MSI2、SERPINE1和JAM - A为miR - 145的靶基因。最后，在显性（AG + GG对AA）和隐性模型（GG对AG + AA）中均观察到显著的遗传效应，表明SNP rs710886的基因型与子宫内膜异位症风险之间存在关联。SNP rs710886 A>G可降低PCAT1的表达，从而导致miR - 145的过表达。高表达的miR - 145将通过靶向特定基因抑制子宫内膜异位症干细胞的侵袭和增殖，从而降低子宫内膜异位症的风险。
