Cai Lily, Chen Juan-Juan, Deng Fu-Mou, Wang Lei, Chen Yu
Department of Clinical Laboratory, The Second Affiliated Hospital of Nanchang University, Jiangxi Provincial Key Laboratory of Laboratory Medicine, Nanchang, China.
Department of Anesthesiology, The Second Affiliated Hospital of Nanchang University, Nanchang, China.
J Obstet Gynaecol Res. 2021 Feb;47(2):621-630. doi: 10.1111/jog.14572. Epub 2020 Nov 18.
MiR-326 has been investigated to be correlated with multiple types of malignancies; however, the role of miR-326 in endometrial cancer (EC) remains rarely reported. The aim of our research is to investigate the functions of miR-326 in EC and the potential molecular mechanism.
RT-qPCR was performed to compare the expression of miR-326 and Bcl-2 in normal endometrial epithelial cell line (End1/e6e7) and EC cells lines (HEC-1A, Ishikawa), respectively. Bioinformatic analysis and luciferase assay verified the relationship between miR-326 and the 3'-UTR of Bcl-2. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, soft agar colony formation assay and the flow cytometry were performed to investigate the functions of miR-326 and Bcl-2 on proliferation and apoptosis in EC. Western blotting was employed to explore the expression of Bcl-2, Bcl2-associated X (Bax) and caspase-3.
The expression of miR-326 decreased in EC cell lines compared to normal endometrial epithelial cell line, while Bcl-2 expression was increased in EC cells. Results of MTT and soft agar colony formation assays showed that miR-326 suppressed proliferation in EC cells. In addition, flow cytometry revealed that miR-326 promoted apoptosis in EC cells. Western blotting showed that silencing miR-326 promoted the expression of Bcl-2. Bioinformatics analysis and luciferase assay verified the 3'-UTR of Bcl-2 was a target of miR-326. Furthermore, MTT assay, soft agar colony formation assay and the flow cytometry proved that miR-326 acts as tumor suppressor via inhibiting the expression of Bcl-2.
MiR-326 acts as a cancer suppressor to inhibit proliferation and promote apoptosis via targeting Bcl-2 axis in EC.
已有研究表明miR-326与多种恶性肿瘤相关；然而，miR-326在子宫内膜癌（EC）中的作用仍鲜有报道。本研究旨在探讨miR-326在EC中的功能及潜在分子机制。
分别采用RT-qPCR比较miR-326和Bcl-2在正常子宫内膜上皮细胞系（End1/e6e7）和EC细胞系（HEC-1A、Ishikawa）中的表达。生物信息学分析和荧光素酶报告基因实验验证miR-326与Bcl-2的3'-UTR之间的关系。采用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐（MTT）实验、软琼脂集落形成实验和流式细胞术研究miR-326和Bcl-2对EC细胞增殖和凋亡的影响。采用蛋白质免疫印迹法检测Bcl-2、Bcl2相关X蛋白（Bax）和半胱天冬酶-3的表达。
与正常子宫内膜上皮细胞系相比，EC细胞系中miR-326表达降低，而EC细胞中Bcl-2表达升高。MTT实验和软琼脂集落形成实验结果显示，miR-326抑制EC细胞增殖。此外，流式细胞术显示，miR-326促进EC细胞凋亡。蛋白质免疫印迹法显示，沉默miR-326可促进Bcl-2表达。生物信息学分析和荧光素酶报告基因实验验证Bcl-2的3'-UTR是miR-326的靶标。此外，MTT实验、软琼脂集落形成实验和流式细胞术证明，miR-326通过抑制Bcl-2表达发挥肿瘤抑制作用。
在EC中，miR-326作为一种抑癌基因，通过靶向Bcl-2轴抑制细胞增殖并促进细胞凋亡。
