miR-181a在卵巢癌细胞中对顺铂的耐药作用

徐丽秀; 美力班·吐尔逊; 克热曼·牙库甫

doi:10.12259/j.issn.2095-610X.S20220131

miR-181a在卵巢癌细胞中对顺铂的耐药作用

doi: 10.12259/j.issn.2095-610X.S20220131

1.
新疆医科大学基础医学院，新疆乌鲁木齐　830001
2.
新疆维吾尔自治区人民医院妇科，新疆乌鲁木齐　830001

基金项目: 新疆高发肿瘤宫颈癌基础研究创新团队基金资助项目（2021D14002）

详细信息

作者简介:
徐丽秀（1992～），女，山东青岛人，在读博士研究生，主要从事妇科肿瘤基础工作

通讯作者:
克热曼·牙库甫，E-mail：771432636@qq.com

中图分类号: R737.31
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- 被引次数: 0
出版历程
- 收稿日期: 2021-12-26
- 网络出版日期: 2021-12-20
- 刊出日期: 2022-01-15

Studies on Cisplatin Resistance of miR-181a in Ovarian Cancer Cells

1.
Basic Medicine College of Xinjiang Medical University，Urumqi Xinjiang，830011
2.
Dept. of Gynecology People’s Hospital of Xinjiang Uygur Autonomous Region，Urumqi Xinjiang，830001

摘要

摘要: 目的探讨miR-181a在卵巢癌化疗抵抗方面的影响及其机制。方法 qRT-PCR检测卵巢癌细胞A2780及A2780/DDP中miR-181a的表达；对A2780及A2780/DDP细胞分别进行miR-181a mimics和inhibitors的转染，并利用qRT-PCR技术验证；MTT法检测转染前后细胞对顺铂的敏感性，利用TargetScan、miRDB与miRwalk 3个MicroRNA靶基因数据库预测miR-181a下游靶点，并通过Western blot技术分析预测基因的表达。结果与A2780细胞相比，miR-181a在A2780/DDP细胞中的表达明显减弱（P < 0.05）。干扰miR-181a表达后，A2780细胞对顺铂的抵抗作用减弱（P < 0.05），而上调miR-181a表达，A2780/DDP细胞对顺铂的抵抗作用增强（P < 0.05）。通过TargetScan、miRDB、miRwalk三个数据库预测到PRKCD可作为miR-181a下游靶基因，下调miR-181a表达可使PRKCD蛋白表达明显增强（P < 0.05）；反之，上调miR-181a表达能够显著抑制PRKCD蛋白表达（P < 0.05）。结论 miR-181a卵巢癌细胞对顺铂的耐药性方面具有抑制作用。
- 卵巢癌 /
- PRKCD /
- miR-181a /
- 顺铂耐药性
Abstract: Objective To investigate the effect of miR-181a on chemotherapy resistance of ovarian cancer and its regulatory mechanism. Methods Real-time PCR was used to detect the expression of miR-181a in A2780 and cisplatin resistant cell lines （A2780/DDP）, and siltation/overexpression changed the expression of miR-181a in A2780 and A2780/DDP cells. MTT method was used to determine the sensitivity of cells to cisplatin before and after the transfection. Three MicroRNA target gene databases, TargetScan, miRDB and miRwalk, were used to predict the downstream targets of miR-181a, and the changes in protein expression of target genes were analyzed by Western blot. Results Compared with A2780 cells, the expression of miR-181a in A2780/DDP cells was significantly decreased （P < 0.05）. After the transfection of A2780 cells with miR-181a inhibitor （interfering with Mir-181A expression）, the sensitivity of A2780 cells to cisplatin decreased （P < 0.05）, and after the transfection of A2780/DDP cells with miR-181a mimic （overexpressing miR-181a）, The sensitivity of cells to cisplatin was increased （P < 0.05）. It was predicted that PRKCD could be used as the downstream target gene of miR-181a by TargetScan, miRDB and miRwalk databases, and down-regulation of miR-181a expression could significantly enhance PRKCD protein expression （P < 0.05）. On the contrary, up-regulation of miR-181a significantly inhibited PRKCD protein expression （P < 0.05）. Conclusion miR-181a may inhibit cisplatin resistance of ovarian cancer cell A2780 by regulating PRKCD.
- Ovarian cancer /
- PRKCD /
- miR-181a /
- Cisplatin resistance

HTML全文

图 1 miR-181a在两种卵巢癌细胞中qRT-PCR结果

与A2780细胞比较，^*P < 0.05。

Figure 1. The expression of miR-181a mRNA in A2780 and A2780/DDP cells

下载: 全尺寸图片幻灯片

图 2 miR-181a在两种细胞中qRT-PCR结果

A：A2780细胞中miR-181a的mRNA相对表达量；B：A2780/DDP细胞中miR-181a的mRNA相对表达量。与空白对照组细胞比较，^*P < 0.05。

Figure 2. The transfection efficiency of miR-181a inhibitors and mimics

下载: 全尺寸图片幻灯片

图 3 miR-181a靶基因预测韦恩图

Figure 3. Venn diagram of screening differentially expressed genes

下载: 全尺寸图片幻灯片

图 4 PRKCD蛋白在两种卵巢癌细胞中Western blot结果

A：A2780细胞中PRKCD的Western blotting结果；B：A2780/DDP细胞中PRKCD的Western blotting结果。

Figure 4. Expression of PRKCD protein in A2780 and A2780/DDP cells

下载: 全尺寸图片幻灯片

表 1 A2780细胞在不同浓度顺铂作用下的MTT数值

Table 1. MTT results of A2780 cells treated with different concentrations of Cisplatin

组别	药物浓度(μmol/L)
组别	0	10	20	30	40	50
阴性对照组	96.48 ± 1.58	73.42 ± 1.82	52.36 ± 1.32	19.87 ± 2.63	11.28 ± 2.36	6.68 ± 1.72
低表达组	97.23 ± 1.22	86.72 ± 1.76*	79.52 ± 1.68*	69.86 ± 1.92*	49.62 ± 1.56*	20.27 ± 1.82*
t	−3.007	−12.026	−130.674	−121.951	−83.009	−235.386
P	0.10	0.01	< 0.001	< 0.001	< 0.001	< 0.001
与阴性对照组比较，^*P < 0.05。

下载: 导出CSV

表 2 A2780/DDP细胞在不同浓度顺铂作用下的MTT数值

Table 2. MTT results of A2780/DDP cells treated with different concentrations of Cisplatin

组别	药物浓度(μmol/L)
组别	0	10	20	30	40	50
阴性对照组	97.42 ± 1.42	92.85 ± 1.72	85.64 ± 1.87	82.48 ± 2.24	76.95 ± 1.98	61.12 ± 2.32
低表达组	97.16 ± 1.82	88.63 ± 1.87	84.72 ± 1.87	76.12 ± 1.87	59.72 ± 2.52	40.58 ± 2.78
t	1.126	48.728	0.6025	29.773	55.265	77.34
P	0.377	< 0.001	0.579	< 0.001	< 0.001	< 0.001

下载: 导出CSV

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