miR-218 functions as a tumor suppressor gene in cervical cancer
miR-218 functions as a tumor suppressor gene in cervical cancer
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DOI：
10.3892/mmr.2019.10809
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发表时间：
2020-01-01
影响因子：
3.4
通讯作者：
Hu, Xiaoxia
中科院分区：
文献类型：
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作者：
Liu, Zhen;Mao, Lin;Hu, Xiaoxia
以往的microRNA(MiR)微阵列分析表明miR-218在宫颈癌组织中表达下调。本研究旨在进一步探讨miR-218在宫颈癌组织中的表达及其与疾病进展的关系，并探讨miR-218在宫颈癌细胞中的作用。组织标本来自80例宫颈鳞癌、30例高度宫颈上皮内瘤变[(CIN)II/III]和15例低度CIN(CINI)，60例宫颈癌患者的血浆标本，15例正常宫颈组织和30例健康妇女的血浆标本。用逆转录定量聚合酶链式反应分析miR-218的表达。另外，将miR-218模拟物、人乳头瘤病毒(HPV)16 E6/E7小干扰RNA或其各自的阴性对照分别导入肿瘤细胞，通过四甲基偶氮唑盐比色法、集落形成实验、伤口愈合实验和Transwell实验检测细胞的活力、集落形成、迁移和侵袭能力。利用基因本体论(GO)术语对miR-218的靶基因进行生物信息学预测和分析。结果表明，miR-218在宫颈癌患者的肿瘤组织和血浆中表达下调，其表达与患者的晚期临床病理特征有关，包括HPV阳性、肿瘤大小、血管侵犯和淋巴结转移。此外，miR-218过表达降低了肿瘤细胞的活力和异种移植瘤的生长，并抑制了肿瘤细胞的迁移和侵袭。80例宫颈癌组织中HPV阳性率为75%，且HPV阳性与miR-218表达呈负相关。此外，生物信息学分析预测，环行引导受体1(Robo1)是miR-218的靶基因；miR-218的过表达显著降低了Robo1的水平。此外，GO分析表明，Robo1参与调控细胞的增殖、黏附和迁移，以及细胞周期。综上所述，本研究结果提示miR-218可能具有抗宫颈癌作用。
Previous microRNA (miR) microarray analysis revealed that miR-218 is downregulated in cervical cancer tissues. The present study aimed to further evaluate the expression of miR-218 in cervical cancer specimens, determine the association between its expression with disease progression, and investigate the roles of miR-218 in cervical cancer cells. Tissue specimens were obtained from 80 patients with cervical squamous cell carcinoma, 30 patients with high-grade cervical intraepithelial neoplasia [(CIN) II/III] and 15 patients with low-grade CIN (CINI); in addition, 60 plasma samples were obtained from patients with cervical cancer, and 15 normal cervical tissue specimens and 30 plasma samples were obtained from healthy women. These samples were used for analysis of miR-218 expression via reverse transcription-quantitative PCR. In addition, tumor cells were transfected with miR-218 mimics, human papillomavirus (HPV)16 E6/E7 small interfering RNA, or their respective negative controls to determine the viability, colony formation, migration and invasion of cells using MTT, colony formation, wound healing and Transwell assays, respectively. Target genes of miR-218 were bioinformatically predicted and analyzed using Gene Ontology (GO) terms. The results revealed that miR-218 was downregulated in the tumor tissues and plasma of patients with cervical cancer, with expression associated with the advanced clinicopathological characteristics of patients, including HPV positivity, tumor size, blood vessel invasion and lymph node metastasis. Furthermore, miR-218 overexpression reduced tumor cell viability and xenograft growth, and suppressed tumor cell migration and invasion. HPV was detected in 75% of the 80 patients with cervical cancer, and HPV positivity was inversely associated with miR-218 expression. In addition, bioinformatics analysis predicted that roundabout guidance receptor 1 (ROBO1) was a target gene of miR-218; miR-218 overexpression significantly reduced ROBO1 levels. Furthermore, GO analysis revealed that ROBO1 was involved in regulating cell proliferation, adhesion and migration, and the cell cycle. In conclusion, the findings of the present study suggested that miR-218 may possess antitumor activities in cervical cancer.