MicroRNA-331-3p Suppresses Cervical Cancer Cell Proliferation and E6/E7 Expression by Targeting NRP2.
MicroRNA-331-3p Suppresses Cervical Cancer Cell Proliferation and E6/E7 Expression by Targeting NRP2.
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DOI：
10.3390/ijms17081351
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发表时间：
2016-08-18
影响因子：
5.6
通讯作者：
Konishi N
中科院分区：
文献类型：
--
作者：
Fujii T;Shimada K;Asano A;Tatsumi Y;Yamaguchi N;Yamazaki M;Konishi N
microRNA（miRNAs）的异常表达参与了多种类型癌症的发生和发展。本研究探讨miR-331- 3 p对宫颈癌细胞增殖及角质形成细胞分化标志物表达的影响。此外，我们评估了神经纤毛蛋白2（NRP 2）是否是调节人乳头瘤病毒（HPV）相关癌蛋白E6和E7的假定靶分子。使用3-（4，5-二甲基噻唑-2-基）-5-（3-羧基甲氧基苯基）-2-（4-磺基苯基）-2H-四唑鎓内盐（MTS）测定评估人宫颈癌细胞系SKG-II、HCS-2和HeLa中的细胞增殖。采用TdT介导的dUTP缺口末端标记法（TUNEL）和Annexin V法检测细胞凋亡。采用定量RT-PCR检测NRP 2、E6、E7、p63和外皮蛋白（IVL）基因的信使RNA（mRNA）表达。使用细胞周期分析进行细胞生长的功能测定。过表达miR-331- 3 p可抑制SKG-II、HCS-2和HeLa细胞的增殖，并诱导细胞发生G2/M期阻滞和凋亡。NRP 2 3′-非翻译区的荧光素酶报告基因分析揭示了miR-331- 3 p对NRP 2的直接调控。在SKG-II、HCS-2和HeLa细胞中使用定量RT-PCR分析过表达miR-331- 3 p或抑制NRP 2的基因表达，发现E6、E7和p63 mRNA下调，IVL mRNA上调。miR-331- 3 p过表达可抑制NRP 2蛋白表达。结果表明，miR-331- 3 p和NRP 2通过调控细胞周期、凋亡等，是细胞增殖的关键效应因子。NRP-2还调节E6/E7和角质形成细胞分化标志物的表达。我们的研究结果表明，miR-331- 3 p在调节宫颈癌细胞增殖中具有重要作用，并且miR-331- 3 p可能通过抑制NRP 2促进角质形成细胞分化。miR-331- 3 p和NRP 2可能有助于抗癌作用。
Aberrant expression of microRNAs (miRNAs) is involved in the development and progression of various types of cancers. In this study, we investigated the role of miR-331-3p in cell proliferation and the expression of keratinocyte differentiation markers of uterine cervical cancer cells. Moreover, we evaluated whether neuropilin 2 (NRP2) are putative target molecules that regulate the human papillomavirus (HPV) related oncoproteins E6 and E7. Cell proliferation in the human cervical cancer cell lines SKG-II, HCS-2, and HeLa was assessed using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. Cellular apoptosis was measured using the TdT-mediated dUTP nick end labeling (TUNEL) and Annexin V assays. Quantitative RT-PCR was used to measure the messenger RNA (mRNA) expression of the NRP2, E6, E7, p63, and involucrin (IVL) genes. A functional assay for cell growth was performed using cell cycle analyses. Overexpression of miR-331-3p inhibited cell proliferation, and induced G2/M phase arrest and apoptosis in SKG-II, HCS-2 and HeLa cells. The luciferase reporter assay of the NRP2 3′-untranslated region revealed the direct regulation of NRP2 by miR-331-3p. Gene expression analyses using quantitative RT-PCR in SKG-II, HCS-2, and HeLa cells overexpressing miR-331-3p or suppressing NRP2 revealed down-regulation of E6, E7, and p63 mRNA and up-regulation of IVL mRNA. Moreover, miR-331-3p overexpression was suppressed NRP2 expression in protein level. We showed that miR-331-3p and NRP2 were key effectors of cell proliferation by regulating the cell cycle, apoptosis. NRP-2 also regulates the expression of E6/E7 and keratinocyte differentiation markers. Our findings suggest that miR-331-3p has an important role in regulating cervical cancer cell proliferation, and that miR-331-3p may contribute to keratinocyte differentiation through NRP2 suppression. miR-331-3p and NRP2 may contribute to anti-cancer effects.
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miR-331-3p调节神经蛋白-2在胶质母细胞瘤中的表达。
影响因子：
3.9
作者：
Epis, Michael R.;Giles, Keith M.;Candy, Patrick A.;Webster, Rebecca J.;Leedman, Peter J.
通讯作者：
Leedman, Peter J.
影响因子：
3.8
作者：
Fujii T;Shimada K;Tatsumi Y;Hatakeyama K;Obayashi C;Fujimoto K;Konishi N
通讯作者：
Konishi N
影响因子：
5.6
作者：
Jung HS;Rajasekaran N;Song SY;Kim YD;Hong S;Choi HJ;Kim YS;Choi JS;Choi YL;Shin YK
通讯作者：
Shin YK
影响因子：
4.8
作者：
Gyöngyösi E;Szalmás A;Ferenczi A;Kónya J;Gergely L;Veress G
通讯作者：
Veress G
影响因子：
3.5
作者：
Gustinucci, Daniela;Rossi, Paolo Giorgi;Passamonti, Basilio
通讯作者：
Passamonti, Basilio