目的： 探索微小RNA-424-5p/驱动蛋白家族成员23（miR-424-5p/KIF23）轴对肝癌细胞恶性表型及其对索拉非尼敏感性的影响。 方法： 实时荧光定量PCR（qRT-PCR）和/或蛋白免疫印迹法检测miR-424-5p和KIF23在肝癌组织及癌旁组织、人肝癌细胞HepG2及正常肝细胞LO2中的表达情况。将分别转染miR-424-5p模拟物（mimic）和miR-424-5p mimic 对照（NC）的HepG2细胞作为miR-424-5p mimic组和mimic NC组；将分别转染KIF23过表达载体pcDNA3.1-KIF23和空载体pcDNA3.1的HepG2细胞作为OE-KIF23组和Vector对照组，将同时转染miR-424-5p mimic和过表达载体pcDNA3.1-KIF23的HepG2细胞作为mimic+OE-KIF23组。将KIF23-3’UTR野生型载体（KIF23-WT）和突变型载体（KIF23-MT）分别与miR-424-5p mimic和mimic NC进行共转染，并用双荧光素酶报告实验验证miR-424-5p与KIF23的靶向关系；细胞计数试剂盒（CCK-8）检测各组HepG2细胞增殖及其对索拉非尼的敏感性；流式细胞术评估各组HepG2细胞凋亡情况；Transwell和划痕实验检测各组HepG2细胞迁移和侵袭能力。组间数据比较采用t检验或方差分析。 结果： 与癌旁组织和正常肝细胞相比，肝癌组织和肝癌细胞的miR-424-5p均显著下调（相对表达量分别为0.604±0.121，0.585±0.064），KIF23显著上调（相对表达量分别为5.451±1.834，2.482±0.545），P值均<0.05。miR-424-5p mimic可抑制肝癌细胞的增殖、迁移和侵袭，促进肝癌细胞的凋亡（P值均<0.05）。过表达KIF23可促进肝癌细胞的增殖、迁移和侵袭，抑制肝癌细胞的凋亡（P值均<0.05）。与mimic NC和KIF23-WT共转染组HepG2细胞相比，mimic和KIF23-WT共转染组HepG2细胞的荧光素酶活性显著降低（相对荧光强度分别为：3.668±0.091、2.629±0.056，P<0.01），而mimic和KIF23-MT共转染组与mimic NC和KIF23-MT共转染组细胞的荧光素酶活性相比，差异无统计学意义。miR-424-5p mimic可逆转过表达KIF23对肝癌细胞增殖、迁移和侵袭能力的促进作用（P值均<0.05）。索拉非尼对mimic+OE-KIF23组和OE-KIF23组HepG2细胞的抑制率分别为47.491%±3.863%和36.246%±6.063%（t=3.027，P<0.05）。 结论： miR-424-5p可通过靶向调控KIF23的表达水平抑制肝癌细胞的增殖、迁移和侵袭，并提高肝癌细胞对索拉非尼的敏感性。.

2022-09-01·Molecular Biology Reports

The combined restoration of miR-424-5p and miR-142-3p effectively inhibits MCF-7 breast cancer cell line via modulating apoptosis, proliferation, colony formation, cell cycle and autophagy

Article

作者: Azarbarzin, Shirin ; Rajabi, Ali ; Dastmalchi, Narges ; Safaralizadeh, Reza ; Khojasteh, Seyed Mahdi Banan ; Hosseinpourfeizi, Mohammad Ali ; Baradaran, Behzad ; Shadbad, Mahdi Abdoli

BACKGROUNDThe combined restoration of tumor-suppressive microRNAs (miRs) has been identified as a promising approach for inhibiting breast cancer development. This study investigated the effect of the combined restoration of miR-424-5p and miR-142-3p on MCF-7 cells and compared the efficacy of the combined therapy with the monotherapies with miR-424-5p and miR-142-3p.METHODSAfter transfection of miR-424-5p and miR-142-3p mimics into MCF-7 cells in the combined and separated manner, the proliferation of tumoral cells was assessed by the MTT assay. Also, the apoptosis, autophagy, and cell cycle of the cells were analyzed by flow cytometry. Western blot and qRT-PCR were used to study the expression levels of c-Myc, Bcl-2, Bax, STAT-3, Oct-3, and Beclin-1.RESULTSOur results have demonstrated that the combined restoration of miR-424-5p and miR-142-3p is more effective in inhibiting tumor proliferation via upregulating Bax and Beclin-1 and downregulating Bcl-2 and c-Myc. Besides, the combined therapy has arrested the cell cycle in the sub-G1 and G2 phases and has suppressed the clonogenicity via downregulating STAT-3 and Oct-3, respectively.CONCLUSIONThe combined restoration of miR-424-5p and miR-142-3p is more effective in inhibiting MCF-7 breast cancer development than monotherapies with miR-424-5p and miR-142-3p.

2021-06-01·Reproductive BioMedicine Online3区 · 医学

Effects of CDKN2B-AS1 on cellular proliferation, invasion and AKT3 expression are attenuated by miR-424-5p in a model of ovarian endometriosis

3区 · 医学

Article

作者: Zhang, Xinyue ; Fang, Xiaoling ; Yi, Mingyu ; Cao, Le ; Jiang, Li ; Wang, Sixue ; Zhang, Tingting ; Zhou, Yuxin

RESEARCH QUESTIONEndometriosis is a common and complicated gynaecologic disease. Long non-coding RNA CDKN2B-AS1 plays a crucial role in the development and progression of several cancers. Whether CDKN2B-AS1 contributes to endometriosis, however, remains unknown.DESIGNCellular proliferation, invasion and DNA synthesis abilities were assessed by CCK8, transwell and 5-ethynyle-2'-deoxyuridine assays. The expression of epithelial-mesenchymal transition markers and three isoforms of AKT was detected using Western blot. Real-time polymerase chain reaction was used to determine the relative expression levels of CDKN2B-AS1 and candidate miRNAs in ectopic, eutopic endometria and normal endometrial tissues. The relationship between CDKN2B-AS1 and miRNA was determined by luciferase reporter assays.RESULTSThe relative expression level of CDKN2B-AS1 was up-regulated in eutopic and ectopic endometria. In endometrial stromal cells and Ishikawa cells, CDKN2B-AS1 overexpression promoted cellular proliferation and invasion, and increased the protein expression of vimentin but decreased the expression of E-cadherin. miR-424-5p was confirmed the target of CDKN2B-AS1 through bioinformatics tools and luciferase reporter assays. In addition, the enhanced effect of cellular phenotype of CDKN2B-AS1 overexpression was significantly attenuated by miR-424-5p overexpression. Furthermore, miR-424-5p was able to directly target AKT3 through luciferase reporter assay. Mechanistically, CDKN2B-AS1 acts as a ceRNA by sponging miR-424-5p and targets AKT3.CONCLUSIONSThe cellular mechanism of CDKN2B-AS1 in endometriosis was confirmed; CDKN2B-AS1 may be a potential target for ovarian endometriosis therapy.

100 项与 miR-424-5p (Kagoshima University) 相关的药物交易

登录后查看更多信息