Tianjin Central Obstetrics and Gynecology Hospital, Tianjin Key Laboratory of Human Development and Reproductive Regulation, Tianjin, 300100, China.
Funct Integr Genomics. 2023 Jul 8;23(3):226. doi: 10.1007/s10142-023-01148-3.
Dysregulation of noncoding RNAs has been reported to have a close correlation with preeclampsia(PE)development. TCL6 was upregulated in patients with PE. In this study, we examined the impacts of TCL6 on modulating HTR-8/SVneo cells induced by LPS. LPS (100 and 200 ng/ml) was applied to induce inflammation in trophoblast cells HTR-8/SVneo. Cell viability, apoptosis, and transwell experiments were conducted. The ELISA methods were used for pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. MDA, GSH, and GPX kits were employed. Transfection was performed for expression regulation of TCL6, miR-485-5p, and TFRC in cells. Bioinformatic online tools were used to predict the targeting sites. Luciferase and RNA immunoprecipitation-qPCR were done to verify the interactions of TCL6, miR-485-5p, and TFRC. RNA expression levels were measured using RT-qPCR, and protein expression of TFRC and GPX4 was detected using a western blot. The free Fe (II) contents were measured. LPS decreased viability, invasion, and migration but enhanced apoptosis, ferroptosis, and inflammation. TCL6 expression was enhanced by LPS induction. The knockdown of TCL6 increased HTR-8/SVneo cell viability and invasion but inhibited cell apoptosis, inflammation, and ferroptosis while inhibition of miR-485-5p could reverse this through TFRC regulation. Moreover, miR-485-5p was sponged by TCL6 and bound to TFRC. TCL6 protected trophoblast cells from LPS-induced injury through the TFRC pathway.
非编码 RNA 的失调已被报道与子痫前期 (PE) 的发展密切相关。TCL6 在患有 PE 的患者中上调。在这项研究中，我们研究了 TCL6 对调节 LPS 诱导的 HTR-8/SVneo 细胞的影响。LPS（100 和 200ng/ml）用于诱导滋养细胞 HTR-8/SVneo 中的炎症。进行细胞活力、凋亡和 Transwell 实验。使用 ELISA 方法检测促炎细胞因子 IL-1β、IL-6 和 TNF-α。使用 MDA、GSH 和 GPX 试剂盒。进行 TCL6、miR-485-5p 和 TFRC 在细胞中的表达调控转染。使用生物信息学在线工具预测靶向位点。进行荧光素酶和 RNA 免疫沉淀-qPCR 以验证 TCL6、miR-485-5p 和 TFRC 的相互作用。使用 RT-qPCR 测量 RNA 表达水平，使用 Western blot 检测 TFRC 和 GPX4 的蛋白表达。测量游离 Fe (II) 含量。LPS 降低了活力、侵袭和迁移，但增强了凋亡、铁死亡和炎症。LPS 诱导 TCL6 表达增加。TCL6 的敲低增加了 HTR-8/SVneo 细胞活力和侵袭，但抑制了细胞凋亡、炎症和铁死亡，而 miR-485-5p 的抑制可通过 TFRC 调节来逆转这种情况。此外，miR-485-5p 被 TCL6 海绵吸附并与 TFRC 结合。TCL6 通过 TFRC 途径保护滋养细胞免受 LPS 诱导的损伤。
