MicroRNA-154-5p regulates the HPV16 E7-pRb pathway in Cervical Carcinogenesis by targeting CUL2
MicroRNA-154-5p regulates the HPV16 E7-pRb pathway in Cervical Carcinogenesis by targeting CUL2
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MicroRNA-154-5p 通过靶向 CUL2 调控 HPV16 E7-pRb 通路在宫颈癌发生中的作用
DOI：
10.7150/jca.45871
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发表时间：
2020-01-01
影响因子：
3.9
通讯作者：
Hao, Min
中科院分区：
文献类型：
--
作者：
Zhao, Weihong;Liu, Yatao;Hao, Min
宫颈癌是由持续的HPV感染引起的，具有很高的死亡率。E3泛素连接酶Cullin 2（CUL 2）对于HPV 16 E7介导的视网膜母细胞瘤蛋白（pRb）降解至关重要。在肿瘤发生过程中会诱导microRNAs（miRNAs）的失调;然而，miRNAs网络与宫颈癌特异性CUL 2之间的关联仍然未知。在此，我们使用AffymiRNA阵列确定了宫颈癌组织中的miRNA谱。我们发现，miR-154- 5 p在130个活检标本中使用实时定量逆转录PCR在癌症进展过程中下调。生物信息学分析和双荧光素酶报告基因分析表明miR-154- 5 p直接靶向CUL 2 3 'UTR。为了确定调节miR-154- 5 p和CUL 2水平的功能结果，用miR-154- 5 p模拟物、miR-154 - 5 p抑制剂或CUL 2 siRNA转染HPV 16阳性宫颈癌细胞系（SiHa）。采用CCK 8细胞计数试剂盒、创伤愈合实验和Transwell侵袭实验检测转染细胞的增殖、迁移和侵袭能力。增加miR-154- 5 p表达促进显著降低的SiHa细胞增殖、迁移和侵袭，而miR-154- 5 p抑制剂具有相反的作用。CUL 2沉默具有与miR-154- 5 p模拟物类似的效果。与miR-154- 5 p和CUL 2水平之间的负相关性一致，CUL 2沉默也增加了pRb表达。据我们所知，这是第一个证明miR-154- 5 p通过靶向CUL 2 3 'UTR调节pRb表达，从而在HPV 16 E7诱导的宫颈癌发生中发挥肿瘤抑制作用的研究。
Cervical cancer, induced by persistent HPV infection, has a high mortality rate. The E3 ubiquitin ligase Cullin 2 (CUL2) is critical for HPV16 E7-mediated degradation of retinoblastoma protein (pRb). Dysregulation of microRNAs (miRNAs) is induced during tumorigenesis; however, the association between miRNA networks and CUL2, specific to cervical cancer, remains unknown. Herein, we determined miRNA profiles in cervical cancer tissues using an Affymetrix miRNA array. We found that miR-154-5p was downregulated during cancer progression using real-time quantitative reverse transcription PCR in 130 biopsy specimens. Bioinformatics analysis and dual-luciferase reporter assays indicated that miR-154-5p directly targets the CUL2 3'UTR. To determine the functional consequences of modulating miR-154-5p and CUL2 levels, HPV16-positive cervical cancer cell line (SiHa) was transfected with miR-154-5p mimic, miR-154-5p inhibitor, or CUL2 siRNA. The proliferation, migration, and invasion of transfected cells were evaluated using CCK8 cell counting kit, wound-healing assay, and Transwell invasion assay. Increased miR-154-5p expression promoted significantly reduced SiHa cell proliferation, migration, and invasion, whereas the miR-154-5p inhibitor had the opposite effect. CUL2 silencing had similar effects to those of the miR-154-5p mimic. Consistent with the inverse correlation between miR-154-5p and CUL2 levels, CUL2 silencing also increased pRb expression. To our knowledge, this is the first study to demonstrate that miR-154-5p regulates pRb expression by targeting CUL2 3'UTR, thereby playing a tumor-suppressive role in HPV16 E7-induced cervical carcinogenesis.