CircRNA EZH2通过调控miR-30c-5p促进前列腺癌细胞增殖和迁移

赵斌; 段元鹏; 张国颖; 毕城伟; 杨李波; 施致裕; 杨勇; 张建朋; 高婷

doi:10.12259/j.issn.2095-610X.S20220731

CircRNA EZH2通过调控miR-30c-5p促进前列腺癌细胞增殖和迁移

doi: 10.12259/j.issn.2095-610X.S20220731

1.
云南省肿瘤医院泌尿外科，云南昆明　650118
2.
云南财经大学校医院，云南昆明　650021

基金项目: 云南省科技厅 - 昆明医科大学应用基础研究联合专项基金资助项目（202001AY070001-069）；云南省教育厅科学研究基金资助项目（2020J0207）

详细信息

作者简介:
赵斌（1976～），男，云南蒙自人，医学硕士，副教授，主要从事泌尿外科肿瘤的研究与临床诊治研究工作

通讯作者:
高婷，E-mail：330155226@qq.com

中图分类号: R737.25
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出版历程
- 收稿日期: 2022-04-27
- 网络出版日期: 2022-06-24
- 刊出日期: 2022-07-14

CircRNA EZH2 Promotes Proliferation and Migration of Prostate Cancer Cells by Regulating miR-30c-5p

1.
Dept. of Urinary Surgery，Yunnan Provincial Tumor Hospital，Kunming Yunnan 650118
2.
University Hospital，Yunnan University of Finance and Economics，Kunming Yunnan 650021，China

摘要

摘要: 目的探究circRNA EZH2通过调控miR-30c-5p对前列腺癌细胞的增殖和迁移的影响及其相关作用机制。方法通过TCGA数据库分析circRNA EZH2在前列腺癌组织中的表达差异及生存曲线；转染siRNA敲降前列腺癌LNCaP细胞中的circRNA EZH2表达后，CCK-8实验和划痕实验观察LNCaP细胞增殖和迁移，RT-qPCR和Western blot法测定miR-30c-5p、JAK1和PI3K蛋白的表达。结果在前列腺癌组织中circRNA EZH2的表达显著上调（P < 0.0001），且高表达circRNA EZH2的患者生存率降低；沉默circRNA EZH2的LNCaP细胞较正常LNCaP细胞的增殖和迁移能力下降（P < 0.0001），miR-30c-5p表达增高（P < 0.0001），JAK1表达降低（P < 0.0001），PI3K信号受抑制。结论 EZH2在前列腺癌中高表达，通过调控miR-30c-5p激活PI3K信号通路促进前列腺癌LNCaP细胞的增殖和迁移。
- 前列腺癌 /
- 环状RNA-EZH2 /
- miR-30c-5p /
- 增殖 /
- 迁移
Abstract: Objective To investigate the effect and related mechanism of circRNA EZH2 on proliferation and migration of prostate cancer cells through miR-30c-5p. Methods The expression difference and survival probability of EZH2 in prostate cancer were analyzed according to TCGA database. SiRNA were transfected to silence EZH2 in prostate cancer LNCaP cells, and proliferation and migration of LNCaP cells were detected by CCK-8 assay and wound healing assay, and the expression of miR-30c-5p, JAK1 and PI3K proteins were determined by RT-qPCR and western blot. Results The expression of EZH2 was up-regulated in prostate cancer tissues, and the survival rate of patients with high expression of EZH2 decreased. Compare with normal LNCaP cells, the proliferation and migration of EZH2-silenced LNCaP cells were decreased, miR-30c-5p expression was increased, JAK1 expression was decreased, and PI3K signal was inhibited. Conclusion EZH2 is highly expressed in prostate cancer and can promote the proliferation and migration of LNCaP cells by regulating miR-30c-5p/JAK1 axis and acting on PI3K signaling pathway.
- Prostate cancer /
- CircRNA EZH2 /
- miR-30c-5p /
- Proliferation /
- Migration

HTML全文

图 1 CircRNA EZH2在前列腺癌中高表达

A：CircRNA EZH2在前列腺癌组织中的表达差异；B：CircRNA EZH2在前列腺癌患者中的生存曲线；C：circRNA EZH2在前列腺癌LNCaP细胞和正常前列腺上皮RWPE1细胞中的表达。^***P < 0.001。

Figure 1. circRNA EZH2 in prostate cancer

下载: 全尺寸图片幻灯片

图 2 qRT-PCR检测LNCaP细胞总RNA经RNase R处理后circRNA EZH2及线性EZH2的表达

^***P < 0.001。

Figure 2. qRT-PCR detected the expression of circRNA EZH2 and linear EZH2 after RNase treatment total RNA from LNCaP cells

下载: 全尺寸图片幻灯片

图 3 CircRNA EZH2对前列腺癌细胞增殖和迁移的影响

A：circRNA EZH2的RT-qPCR结果分析图；B：CCK-8实验结果；D：LNCaP细胞生长情况图；C和E：LNCaP细胞划痕实验及其结果分析。^****P < 0.0001。

Figure 3. The effects of circRNA EZH2 on proliferation and migration of prostate cancer cells

下载: 全尺寸图片幻灯片

图 4 CircRNA EZH2靶向miR-30c-5p

A：circRNA EZH2和miR-30c-5p的配对序列；B：双荧光素酶报告基因检测WT/MUT-circRNA EZH2插入的报告基因和miR-30c-5p mimic及其阴性对照共转染LNCaP细胞后的荧光素酶活性；C：RT-qPCR实验检测miR-30c-5p抑制剂的干扰效果；D：miR-30c-5p的RT-qPCR结果分析；E：circRNA EZH2的RT-qPCR结果分析。^**P < 0.01；^***P < 0.001；^****P < 0.0001。

Figure 4. CircRNA EZH2 targeted miR-30c-5p

下载: 全尺寸图片幻灯片

图 5 CircRNA EZH2调控miR-30c-5p轴对前列腺癌细胞增殖和迁移的影响

A：CCK-8实验结果；B：LNCaP细胞生长情况图； C和D：LNCaP细胞划痕实验及其结果分析。^*P < 0.05；^****P < 0.0001。

Figure 5. CircRNA EZH2 regulated cell proliferation and migration via miR-30c-5p in prostate cancer cell

下载: 全尺寸图片幻灯片

图 6 CircRNA EZH2对JAK1和PI3K的调控

A：JAK1的RT-qPCR结果分析；B-D：JAK1和PI3K的western blot及其结果分析。^*P < 0.05；^**P < 0.01；^***P < 0.001；^****P < 0.0001。

Figure 6. CircRNA EZH2 regulated JAK1 and PI3K

下载: 全尺寸图片幻灯片

表 1 RT-qPCR所需引物序列

Table 1. Primer sequences

基因	引物序列（5′-3′）
circRNA EZH2	Forward Primer：GGACCACAGTGTTACCAGCAT
	Reversed Primer：GTGGGGTCTTTATCCGCTCAG
miR-30c-5p	Forward Primer：GCAGTGTAAACATCCTACACTCT
	Reversed Primer：TCCAGTTTTTTTTTTTTTTTGCTGA
JAK1	Forward Primer：ATGTCCTTGTTGAGAGTGA
	Reversed Primer：CATCCTTGACGGTGTAATAC
GAPDH	Forward Primer：TGTGGGCATCAATGGATTTGG
	Reversed Primer：ACACCATGTATTCCGGGTCAAT
U6	Forward Primer：GCTTCGGCAGCACATATACTAAAAT
	Reversed Primer：CGCTTCACGAATTTGCGTGTCAT

下载: 导出CSV

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