Background: Accumulating documents have demonstrated that long noncoding RNAs (lncRNAs) play critical roles in tumorigenesis. As an lncRNA, nuclear-enriched abundant transcript 1 (NEAT1) has been identified to be involved in the progression of many types of cancers. However, the biological function of NEAT1 in cervical cancer is not fully investigated. The aim of this study was to disclose the specific biological function of lncRNA NEAT1 in cervical cancer progression.
Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was utilized to identify the expression of lncRNA NEAT1 in the cervical cancer tissues and cell lines. All cervical cancer samples used in this study were collected from the Affiliated Suzhou Hospital of Nanjing Medical University between September 2012 and September 2017. The correlation between NEAT1 expression and the overall survival rate of cervical cancer patients was analyzed by Kaplan-Meier analysis. The effects of NEAT1 knockdown or overexpression on cell proliferation were tested by performing MTT assays and colony formation assays. Transwell assays were conducted to detect the migratory ability of cervical cancer cells, in which NEAT1 was silenced or overexpressed. Western blotting was utilized to validate whether NEAT1 promotes cervical cancer progression through activating PI3K-Akt signaling pathway.
Results: High expression of NEAT1 predicted poor prognosis of cervical cancer patients (χ2 = 0.735, P = 0.005). Knockdown of NEAT1 decreased the number of colonies in CaSki cell from 136.667 ± 13.503 to 71.667 ± 7.506 (t = -18.76, P = 0.003) and decreased the number of colonies in HeLa cell from 128.667 ± 13.317 to 65.667 ± 7.024 (t = -5.54, P = 0.031). However, overexpression of NEAT1 increased the number of colonies in SiHa cell from 84.667 ± 12.014 to 150.667 ± 18.037 (t = 7.27, P = 0.018). Knockdown of NEAT1 decreased the migratory number of CaSki cell from 100.333 ± 9.866 to 58.333 ± 5.859 (t = -8.08, P = 0.015) and reduced the migratory number in HeLa cell from 123.667 ± 12.097 to 67.667 ± 7.095 (t = -6.03, P = 0.026). Overexpression of NEAT1 increased the migratory number of SiHa cell from 127.333 ± 16.042 to 231.333 ± 31.786 (t = 4.92, P = 0.039).
Conclusion: NEAT1 may exert oncogenic function in cervical cancer and serve as a novel therapeutic target for cervical cancer.
长链非编码RNA NEAT1在宫颈癌发生中的生物学作用及其机制研究摘要背景：越来越多的研究表明长链非编码RNA能够在肿瘤形成中发挥重要作用。长链非编码RNA NEAT1 已被证实可参与多种癌症的发生发展。然而，NEAT1在宫颈癌中的生物学功能未有相关报道。本研究旨在揭露长链非编码RNA NEAT1在宫颈癌的癌症发生发展过程中的生物学作用及其机制。 方法：研究中所有组织样本均从南京医科大学附属苏州医院获取。样本收集时间为2012年9月到2017年9月。NEAT1的表达与患者总体生存率间的关系由Kaplan-Meier生存曲线分析得出。MTT和克隆实验方法检测NEAT1干涉和过表达对细胞增殖能力的影响。在NEAT1被干涉或过表达后，Transwell转移实验被用于检测宫颈癌细胞的转移能力。蛋白免疫印迹方法检测PI3K-Akt信号通路相关蛋白的表达水平。转染了si-NC或pcDNA-NC的癌细胞株作为阴性对照。 结果：NEAT1的高表达预示了宫颈癌病人的不良预后(χ2 = 7.735, P =0.005）。干涉NEAT1 将CaSki细胞的活性从136.667 ± 13.503降低到71.667 ± 7.506(t = -18.76, P = 0.003)将HeLa细胞的活性从128.667 ± 13.317降低到65.667±7.024(t = -5.54, P = 0.031)。然而过表达NEAT1将SiHa细胞的活性84.667 ± 12.014升高到150.667 ± 18.037(t = 7.27,P = 0.018)。干涉NEAT1后，CaSki细胞的转移数量从100.333 ± 9.866降低到58.333 ± 5.859(t = -8.08, P = 0.015)，HeLa细胞的转移数量从123.667 ± 12.097降低到67.667 ± 7.095(t = -6.03, P = 0.026)。NEAT1过表达使SiHa细胞的转移数量从127.333 ± 16.042增长到231.333 ± 31.786 (t = 4.92, P = 0.039)。 结论：NEAT1在宫颈癌发生发展中可能发挥了癌基因的作用，NEAT1可能是宫颈癌的一个新的治疗靶点。.
Keywords: Cervical Cancer; Long Noncoding RNA Nuclear-Enriched Abundant Transcript 1; Migration; PI3K/Akt Signaling Pathway; Proliferation.