Liu Y-J, Zhou H-G, Chen L-H, Qu D-C, Wang C-J, Xia Z-Y, Zheng J-H
Department of Obstetrics and Gynecology, Affiliated Hospital of North Sichuan Medical College, Nanchong, China.
Eur Rev Med Pharmacol Sci. 2019 Jan;23(1):87-95. doi: 10.26355/eurrev_201901_16752.
To investigate the effects of miR-32-5p on the biological behaviors of cervical cancer (CCa), the relevant mechanism was studied in CCa cell lines (HeLa) in vitro.
The expression level of miR-32-5p was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). TargetScan, miRDB, microRNA databases and Luciferase method were conducted to predict and validate the target gene of miR-32-5p; the effects of miR-32-5p on cell proliferation, clone formation, invasion and migration capacity were analyzed in vitro study.
We found miR-32-5p to be significantly inhibited in CCa tissues and cells. Bioinformatics approach together with Luciferase method screened Homeobox B8 (HOXB8) as a downstream regulatory target of miR-32-5p. Besides, HOXB8 was incredibly high expression in CCa tissues and cells. After transfection in HeLa cells by miR-32-5p mimics, HOXB8 expression was indicated to be negatively correlated with miR-32-5p both in qRT-PCR and Western blot (WB) assays. The subsequent experiments showed that decreased expression of HOXB8 resulting from up-regulation of miR-32-5p could weaken the cell proliferation, clone formation, invasion and migration ability of HeLa cells.
MiR-32-5p could inhibit the cellular malignant behavior through regulating the expression of HOXB8 in HeLa cells. We provide a new clue for the study of molecular mechanisms of CCa. MiR-32-5p/HOXB8 axis might serve as potential target for the clinical diagnosis and treatment of CCa.
研究miR - 32 - 5p对宫颈癌(CCa)生物学行为的影响,在体外CCa细胞系(HeLa)中研究其相关机制。
采用定量逆转录聚合酶链反应(qRT - PCR)检测miR - 32 - 5p的表达水平。利用TargetScan、miRDB、microRNA数据库及荧光素酶法预测并验证miR - 32 - 5p的靶基因;通过体外实验分析miR - 32 - 5p对细胞增殖、克隆形成、侵袭和迁移能力的影响。
我们发现miR - 32 - 5p在CCa组织和细胞中显著下调。生物信息学方法结合荧光素酶法筛选出同源盒B8(HOXB8)作为miR - 32 - 5p的下游调控靶点。此外,HOXB8在CCa组织和细胞中高表达。用miR - 32 - 5p模拟物转染HeLa细胞后,qRT - PCR和蛋白质免疫印迹(WB)分析均表明HOXB8的表达与miR - 32 - 5p呈负相关。随后的实验表明,miR - 32 - 5p上调导致HOXB8表达降低,可削弱HeLa细胞的增殖、克隆形成、侵袭和迁移能力。
miR - 32 - 5p可通过调节HeLa细胞中HOXB8的表达抑制细胞恶性行为。我们为CCa分子机制的研究提供了新线索。miR - 32 - 5p/HOXB8轴可能成为CCa临床诊断和治疗的潜在靶点。