miR-221在TWIST2调控下通过ARID1A和Wnt/β-catenin信号通路参与宫颈癌侵袭转移的分子机制

MicroRNAs, the regulatory molecules of cell function, played crucial roles in tumor progression. Our preliminary screening studies have found miR-221 to be a miRNA links to cervical cancer metastasis, which significantly promotes the invasion of cervical cancer cells in vitro. However, the underlying mechanism is still elusive. Based upon bioinformatics, the targets of miR-221 regulation in the upstream and downstream pathways were predicted and observed the highest estimates in the TWIST2, ARID1A and Wnt pathways. Preliminary experiments have verified that over-expression of TWIST2 up-regulates the expression of miR-221 and miR-221 induces the loss of ARID1A. In addition, loss of ARID1A and chromosome translocation of Wnt signaling molecules jointly participate the metastasis of malignant tumors. Therefore, we proposed a novel mechanism that, under the regulation of TWIST2 transcription, miR-221 accelerated the invasion and metastasis of cervical cancer via target gene ARID1A and Wnt signaling pathway. This study was designed to utilize cervical cancer cells matched by metastasis potential and visualized animal models combined with molecular pathologic and molecular biological techniques to explore viable evidence supporting the role of miR-221 promoting cervical cancer metastasis, elucidate the molecule mechanism of the biological function of miR-221 and provide new targets for early prognosis, diagnosis and treatment of invasion and metastasis of cervical cancer in clinical settings.

miRNAs作为细胞功能调控分子在肿瘤进展中发挥重要作用。miR-221是我们前期经筛选获得的宫颈癌转移相关miRNA，显著促进宫颈癌细胞的体外侵袭能力，其作用机制尚不清楚。利用生物信息学预测miR-221的上下游调控靶点，发现TWIST2、ARID1A和Wnt信号通路预测值最高。预实验证实宫颈癌细胞中过表达TWIST2使miR-221表达上调， miR-221诱导ARID1A表达缺失，而文献报道ARID1A缺失与Wnt信号分子染色体移位共同参与肿瘤转移。由此我们提出miR-221在TWIST2转录调控下，通过靶基因ARID1A和Wnt信号通路促进宫颈癌侵袭转移的新机制。本项目拟利用转移潜能高低配对宫颈癌细胞和可视化动物模型结合分子病理学和分子生物学技术，获得miR-221促进宫颈癌转移的可靠依据，阐明miR-221发挥生物学功能的分子机制，为临床早期预测、诊断和治疗宫颈癌转移提供新靶点。

miRNAs作为细胞功能调控分子在肿瘤进展中发挥重要作用。miR-221-3p是我们前期经筛选获得的宫颈癌转移相关miRNA，能显著促进宫颈癌细胞的体外侵袭能力，然而其作用机制尚不清楚。本项目通过建立有效实验模型，从组织、细胞、亚细胞、分子等多个层面，采用原位杂交、双荧光素酶、免疫荧光技术、miRNAs芯片、宫颈癌转移动物模型等技术手段探讨了TWIST2/miR-221-3p/THBS2通路促进宫颈癌侵袭转移的机制。在此基础上，我们还深入研究了HPV整合感染在宫颈癌上皮恶性转化发生中的作用，探讨了肿瘤微环境调节网络促进宫颈癌侵袭转移的分子机制。结果显示：1）miR-221-3p是TWIST2的靶向miRNA，而miR-221-3p的过表达可以抑制THBS2，从而促进宫颈癌细胞发生转移；2）确定了高危HPV在人类基因组中的关键整合位点及其对人类基因组DNA的影响；3）在宫颈癌肿瘤微环境中，TGF-β、肿瘤相关巨噬细胞、外泌体均可通过特有机制促进淋巴结转移。我们的研究深入剖析了宫颈癌疾病进展的生物学特征，提供了新的侵袭转移预测靶点并探索基于分子机制的靶向治疗新措施。为补充宫颈癌的早期防止措施、实现降低我国宫颈癌发病率和死亡率的目的提供了实验数据。