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投稿时间:2022-10-28 网络发布日期:2023-07-19
投稿时间:2022-10-28 网络发布日期:2023-07-19
中文摘要: 目的 探讨miR-219a-5p对子宫内膜癌(EC)细胞增殖、凋亡、迁移、侵袭的影响及其作用机制。方法 对人EC细胞AN3CA进行培养和质粒转染,分为对照组(正常培养)、NC组(转染NC质粒)、miR-219a-5p组(转染miR-219a-5p质粒)、 miR-219a-5p组+激活剂组(转染miR-219a-5p质粒+含PI3K激活剂740 Y-P的培养基培养)。MTT、流式细胞术、划痕实验、Transwell实验分别检测各组细胞增殖、凋亡、迁移能力和侵袭能力; Western Blot检测各组细胞磷酯酰肌醇-3-激酶(PI3K)、蛋白激酶B(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)表达水平。对来自患者的EC和癌旁组织进行miR-219a-5p及PI3K信号通路mRNA表达的检测和验证。结果 在四组细胞培养24、48、72h时miR-219a-5p组细胞增殖活力均最低(F=18.25, 16.39, 18.96, P<0.01);与对照组相比,miR-219a-5p组细胞凋亡率显著增加、细胞迁移和侵袭能力显著降低(P<0.01),PI3K、AKT和mTOR表达水平显著下调(P<0.01);与miR-219a-5p组相比,miR-219a-5p+激活剂组细胞上述指标均呈反向变化(P<0.05);而NC组与对照组上述指标比较差异均无统计学意义(P>0.05)。与癌旁组织相比,癌组织miR-219a-5p表达水平显著降低,PI3K、AKT和mTOR的mRNA表达显著上调(P<0.05)。结论 miR-219a-5p可以抑制EC细胞的增殖、迁移和侵袭能力,促进细胞凋亡,其机制可能与PI3K信号通路活性的抑制相关。
中文关键词: 微小RNA-219a-5p 人子宫内膜癌细胞 凋亡 迁移 侵袭 磷酯酰肌醇-3-激酶信号通路
Abstract:Objective To investigate the effects of miR-219a-5p on proliferation, apoptosis, migration and invasion of endometrial cancer cells and its mechanism.
Methods The human endometrial cancer cell line AN3CA were cultured and transfected with plasmid, and divided into control group(normal culture), NC group(transfected with NC plasmid), miR-219a-5p group(transfected with miR-219a-5p plasmid), miR-219a-5p group+activator group(transfected with miR-219a-5p plasmid+medium containing PI3K activator 740 Y-P).The cell proliferation, apoptosis, migration and invasion of each group were detected by MTT, flow cytometry, scratch assay and Transwell assay. The protein expression levels of phosphoinositide-3-kinase(PI3K), protein kinase B(AKT) and mammalian rapamycin target protein(mTOR) were detected by Western Blot. The mRNA expressions of miR-219a-5p and PI3K signaling pathway in endometrial cancer and adjacent tissues from patients were detected and verified.
Results When the four groups of cells were cultured for 24, 48 and 72 h, the cell proliferation activity of miR-219a-5p group was the lowest (F=18.25, 16.39, 18.96, P<0.01). Compared with control group, the proportion of apoptosis and cell migration increased, and invasion decreased significantly(P<0.01), while the expression levels of PI3K, AKT, and mTOR were significantly downregulated in miR-219a-5p group(P<0.01). Compared with miR-219a-5p group, the above indicators in the miR-219a-5p+activator group showed reverse changes(P<0.05). There was no statistically significant difference in the above indicators between the NC group and the control group(P>0.05). Compared with adjacent tissues, the expression level of miR-219a-5p in cancer tissue was significantly reduced, while the mRNA expression of PI3K, AKT, and mTOR was significantly upregulated(P<0.05).
Conclusion miR-219a-5p can inhibit the proliferation, migration, invasion and apoptosis of endometrial cancer cells, and its mechanism may be related to the inhibition of PI3K signaling pathway activity.
keywords: miR-219a-5p Human endometrial cancer cell Apoptosis Migration Invasion Phosphoinositide-3-kinase signaling pathway
文章编号: 中图分类号:R737.33 文献标志码:A
基金项目:河北省医学科学研究重点课题计划(20200264)
作者 | 单位 |
刘雪洁1 | 1. 保定市第二中心医院妇科,河北 保定 072750; |
高秀娟2 | 2. 承德医学院附属医院妇科,河北 承德 067000; |
李晓妍1 | 1. 保定市第二中心医院妇科,河北 保定 072750 |
Author Name | Affiliation |
LIU Xuejie*, GAO Xiujuan, LI Xiaoyan | *Department of Gynecology, Baoding Second Central Hospital, Baoding, Hebei 072750, China |
引用文本:
刘雪洁,高秀娟,李晓妍.miR-219a-5p对子宫内膜癌细胞增殖、凋亡、侵袭的影响及与PI3K信号通路的关系[J].中国临床研究,2023,36(7):977-982.
刘雪洁,高秀娟,李晓妍.miR-219a-5p对子宫内膜癌细胞增殖、凋亡、侵袭的影响及与PI3K信号通路的关系[J].中国临床研究,2023,36(7):977-982.