MiR-4524b-5p/WTX/β-catenin axis functions as a regulator of metastasis in cervical cancer

Cervical cancer is the second most deadly gynecological tumor worldwide. MicroRNAs (miRNAs) play very important roles in tumor oncogenesis and progression. The mechanism of post-transcription regulation of WTX gene is still unknown. A series of differential miRNAs were discovered by microarray analysis comparing three pairs of primary cervical cancer specimens and their relapsed tumors from three patients. Quantitative reverse transcriptase PCR (qRT-PCR), Western Blot (WB) and Immunohistochemistry (IHC) was used to detect the expression of miR-4524b-5p and WTX in cervical cell lines and tissues. The biological function of miR-4524b-5p and WTX was investigated through knockdown and overexpression with inhibitor/siRNA and mimic/plasmid in vitro and in vivo. In this study, we found that miR-4524b-5p is highly expressed in relapsed cervical cancer specimens. Combined in vitro and in vivo experiments, showed that miR-4524b-5p could regulate the migration and invasion ability of cervical cancer. Furthermore, we also found that miR-4524b-5p could regulate the migration and invasion of cervical cancer by targeting WTX and that WTX could regulate the expression of β-catenin. Taken together, our data identified a miR-4524b-5p/WTX/β-catenin regulatory axis for cervical cancer, and miR-4524b-5p may be a potential target for cervical cancer therapy.宫颈癌是全球范围内第二大致命妇科肿瘤。微小RNA（microRNAs，miRNAs）在肿瘤发生与进展过程中发挥至关重要的作用。目前，WTX基因的转录后调控机制仍未明确。本研究通过微阵列分析，对比3名患者的3对原发性宫颈癌标本及其对应复发肿瘤，筛选得到一系列差异表达的miRNAs。采用定量反转录聚合酶链反应（quantitative reverse transcriptase PCR，qRT-PCR）、蛋白质印迹（Western Blot，WB）与免疫组化（Immunohistochemistry，IHC）技术，检测miR-4524b-5p与WTX在宫颈细胞系及组织中的表达水平。通过体外及体内实验，分别利用抑制剂/siRNA和模拟物/质粒对靶基因进行敲低与过表达，探究miR-4524b-5p与WTX的生物学功能。本研究发现，miR-4524b-5p在复发宫颈癌标本中呈高表达状态。结合体外与体内实验结果证实，miR-4524b-5p可调控宫颈癌细胞的迁移与侵袭能力。进一步研究表明，miR-4524b-5p可通过靶向WTX调控宫颈癌的迁移侵袭过程，且WTX可调控β-连环蛋白（β-catenin）的表达。综上，本研究明确了一条针对宫颈癌的miR-4524b-5p/WTX/β-catenin调控轴，且miR-4524b-5p有望成为宫颈癌治疗的潜在靶点。