miR-429在上皮性卵巢癌进展中的作用及机制研究

赵平鸽; 张亚楠; 张文雅; 徐鹏; 黄敏; 李朦; 滕佳茜; 匡野

doi:10.16766/j.cnki.issn.1674-4152.003194

miR-429在上皮性卵巢癌进展中的作用及机制研究

doi: 10.16766/j.cnki.issn.1674-4152.003194

赵平鸽¹,
张亚楠²,
张文雅³,
徐鹏¹,
黄敏⁴,
李朦¹,
滕佳茜¹,
匡野^1, ,

1.
哈尔滨医科大学附属第二医院妇产科，黑龙江哈尔滨 150086
2.
河北北方学院附属第一医院妇产科，河北张家口 050051
3.
沧州市中心医院妇产科，河北沧州 061011
4.
哈尔滨医科大学医学遗传学研究室，黑龙江哈尔滨 150081

基金项目:

黑龙江省自然科学基金项目 LH2021H034

详细信息

通讯作者:
匡野，E-mail：kuangye-blue@163.com

中图分类号: R737.31 R730
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- 文章访问数: 335
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- 被引次数: 0
出版历程
- 收稿日期: 2022-09-06
- 网络出版日期: 2023-11-23

Role and mechanism of miR-429 in the progression of epithelial ovarian cancer

1.
Department of Obstetrics and Gynecology, the Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, China

摘要

摘要: 目的探讨miR-429在上皮性卵巢癌组织中的表达及其与卵巢癌细胞增殖和迁移的关系，寻找其下游靶基因，并探讨miR-429靶基因DNAJB6在卵巢癌中的作用。方法收集2019年9月—2022年2月期间于哈尔滨医科大学附属第二医院经妇科手术切除得到的27例上皮性卵巢癌组织和14例正常卵巢组织，使用实时荧光定量聚合酶链反应检测组织中miR-429及DNAJB6 mRNA的表达。在人卵巢癌细胞株A2780中敲低miR-429, 使用CCK-8、平板克隆形成及Transwell细胞迁移实验在体外检测肿瘤细胞增殖及迁移能力的变化。采用生物信息学软件预测miR-429下游靶基因并分析其在卵巢癌中的预后价值，应用qRT-PCR和Western blotting实验进一步证实miR-429与DNAJB6之间的调控关系。结果与正常卵巢组织[0.886(0.785，2.014)]相比，miR-429在上皮性卵巢癌组织[2.048(0.830，5.281)]中高表达(P=0.018)，过表达miR-429可以增强卵巢癌细胞的增殖和迁移能力。与正常卵巢组织(1.290±0.781)相比，DNAJB6在上皮性卵巢癌组织(0.710±0.456)中低表达(P=0.004)，与卵巢癌患者的预后关系密切。相对于对照组DNAJB6 mRNA的表达量(1.020±0.251)，敲低miR-429可以上调DNAJB6 mRNA的表达量(2.839±0.305，P=0.001)；敲低miR-429同样也上调了DNAJB6蛋白的表达(P=0.049)。结论 miR-429在上皮性卵巢癌中发挥促癌作用，可以调控靶基因DNAJB6的表达，提示miR-429/DNAJB6可能是卵巢癌治疗的新靶点。
- 上皮性卵巢癌 /
- 微小RNA-429 /
- 增殖 /
- 迁移
Abstract: Objective To investigate the expression of miR-429 in epithelial ovarian cancer tissues and its relationship with the proliferation and migration of ovarian cancer cells, and to explore the role of miR-429 target gene DNAJB6 in ovarian cancer. Methods Twenty-seven epithelial ovarian cancer tissues and fourteen normal ovarian tissues obtained by gynecological surgical resection at the Second Affiliated Hospital of Harbin Medical University during the period from September 2019 to February 2022 were selected to detect the expression of miR-429 and DNAJB6 mRNA in the tissues using real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). In human ovarian cancer cell line A2780, miR-429 was knocked down and changes in tumor cell proliferation and migration ability were measured in vitro by using CCK-8 kit, plate clone formation and Transwell cell migration assay. The downstream target genes of miR-429 were predicted by bioinformatics software and its prognostic value in ovarian cancer was analyzed. RT-qPCR and Western blot assays were applied to further confirm the regulatory relationship between miR-429 and DNAJB6. Results Compared with normal ovarian tissues 0.886(0.785, 2.014), miR-429 was highly expressed in epithelial ovarian cancer tissues 2.048(0.830, 5.281), P=0.018, and overexpression of miR-429 could enhance the proliferation and migration of ovarian cancer cells. Compared with normal ovarian tissues (1.290±0.781), DNAJB6 was lowly expressed in epithelial ovarian cancer tissues (0.710±0.456, P=0.004), which was closely related to the prognosis of ovarian cancer patients. Knockdown of miR-429 upregulated DNAJB6 mRNA expression (2.839±0.305, P=0.001) compared to the control (1.020±0.251); knockdown of miR-429 also upregulated DNAJB6 protein expression (P=0.049). Conclusion miR-429 plays a pro-cancer role in epithelial ovarian cancer and can regulate the expression of the target gene DNAJB6, suggesting that miR-429/DNAJB6 may be a new target for ovarian cancer therapy.
- Epithelial ovarian cancer /
- miR-429 /
- Proliferation /
- Migration

HTML全文

图 1 敲低miR-429对A2780细胞生物学特性的影响

注：A为CCK-8实验结果；B为平板克隆形成实验结果；C为Transwell实验结果。

Figure 1. Effect of miR 429 knockdown on the biological characteristics of A2780 cells

下载: 全尺寸图片幻灯片

图 2 miR-429靶基因预测和卵巢癌患者预后分析

注：A为miR-429预测靶基因韦恩图；B为miRTarBase数据库预测miR-429与DNAJB6的结合位点；C为Kaplan-Meier Plotter数据库显示DNAJB6与卵巢癌患者的生存预后关系。

Figure 2. Prediction of miR-429 target genes and prognostic analysis in ovarian cancer patients

下载: 全尺寸图片幻灯片

图 3 蛋白质印迹实验结果

Figure 3. Western blotting results

下载: 全尺寸图片幻灯片

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