miRNA-15a/16调控Bmi-1蛋白在卵巢癌顺铂化疗耐药中的作用

廉阳秧; 岳红萍; 端娅; 胡红文; 罗芳

doi:10.12259/j.issn.2095-610X.S20231204

miRNA-15a/16调控Bmi-1蛋白在卵巢癌顺铂化疗耐药中的作用

doi: 10.12259/j.issn.2095-610X.S20231204

云南省第三人民医院妇科，云南昆明 650200

基金项目: 云南省科技厅科技计划基金资助项目（2019FD092）

详细信息

作者简介:
廉阳秧（1989～），女，河南焦作人，医学硕士，主治医师，主要从事妇科肿瘤研究工作

通讯作者:
罗芳，E-mail：km2004lf@yeah.net

中图分类号: R737.3
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- 被引次数: 0
出版历程
- 收稿日期: 2023-07-01
- 网络出版日期: 2023-12-19
- 刊出日期: 2023-12-25

The Role of miRNA-15a/16 in Regulating Bmi-1 Protein in Ovarian Cancer Resistance to Cisplatin Chemotherapy

Dept. of Gynecology，The 3rd People’s Hospital of Yunnan Province，Kunming Yunnan 650200，China

摘要

摘要: 目的探讨miRNA-15a和miRNA-16在逆转卵巢癌顺铂化疗耐药过程中的作用机制。方法使用miRNA-15a和miRNA-16模拟物转染人卵巢癌顺铂耐药细胞株CoC1/DDP，予顺铂处理，qRT-PCR检测正常培养CoC1/DDP细胞组、顺铂处理组、阴性对照组、转染miRNA-15a组、转染miRNA-16组和过表达的Bmi-1质粒的转染miRNA-15a和miRNA-16组的miRNA-15a、miRNA-16的表达水平，同时应用Western blot检测各组细胞中Bmi-1的表达水平，CCK-8法、Annexin V/PI法检测每组细胞的存活及凋亡情况、γ-H2AX免疫荧光检测细胞凋亡情况。结果卵巢癌顺铂耐药细胞株CoC1/DDP中miRNA-15a和miRNA-16低表达，Bmi-1蛋白高表达。过表达miRNA-15a、miRNA-16水平，Bmi-1蛋白下降（P < 0.05），细胞存活率下降（P < 0.05），DNA凋亡明显增加（P < 0.05），DNA损伤程度更严重（P < 0.05）。过表达Bmi-1质粒可提高细胞活度（P < 0.05），降低细胞凋亡率（P < 0.05）。结论 Bmi-1蛋白可能是miRNA-15a和miRNA-16的调节蛋白靶点，高表达miRNA-15a和miRNA-16可以通过降低Bmi-1蛋白来提高顺铂对卵巢癌细胞的敏感性。通过对卵巢癌顺铂化疗耐药性分子标记的预测和对卵巢癌肿耐药性目标的攻克提供了新的思路。
- 卵巢癌 /
- miRNA-15a/16 /
- Bmi-1蛋白 /
- 化疗耐药 /
- 肿瘤恶性行为
Abstract: Objective To investigate the mechanisms of miRNA-15a and miRNA-16 in the process of reversing cisplatin resistance in ovarian cancer. Methods Human ovarian cancer cisplatin-resistant cell lines CoC1/DDP were transfected with miRNA-15a and miRNA-16 mimics and treated with cisplatin. qRT-PCR was used to detect the expression levels of miRNA-15a and miRNA-16 in normal CoC1/DDP cell group, cisplatin treated group, negative control group, miRNA-15a transfected group, miRNA-16 transfected group and overexpressed Bmi-1 plasmid. Western blot was used to detect the expression level of Bmi-1 in each group, CCK-8 and Annexin V/PI were used to detect cell survival and apoptosis, and γ-H2AX immunofluorescence was used to detect cell apoptosis. Results The CoC1/DDP ovarian cancer cell line shows low expression of miRNA-15a and miRNA-16, and high expression of Bmi-1 protein, which makes it resistant to cisplatin. When the levels of miRNA-15a and miRNA-16 are overexpressed, the Bmi-1 protein decreases（P < 0.05）, leading to a decrease in cell survival rate（P < 0.05）, a significant increase in DNA apoptosis（P < 0.05）, and more severe DNA damage（P < 0.05）. Overexpression of Bmi-1 plasmid can increase cell viability（P < 0.05） and reduce the rate of cell apoptosis（P < 0.05）. Conclusion The Bmi-1 protein may be a target for the regulation of miRNA-15a and miRNA-16, and overexpression of miRNA-15a and miRNA-16 can increase the sensitivity of ovarian cancer cells to cisplatin by reducing the Bmi-1 protein. This provides a new idea for predicting molecular markers of cisplatin resistance in ovarian cancer and overcoming drug resistance targets in ovarian cancer.
- Ovarian cancer /
- miRNA-15a/16 /
- Bmi-1 protein /
- Chemotherapy resistance /
- Malignant behavior of tumor

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图 1 不同浓度顺铂处理的细胞活度

Figure 1. Cell activity was treated at different concentrations of cisplatin

下载: 全尺寸图片幻灯片

图 2 miRNA-15a在各组细胞中的表达水平

与NC组比较，^*P < 0.05。

Figure 2. The expression level of miRNA15a in each group of cells

下载: 全尺寸图片幻灯片

图 3 miRNA-16在各组细胞中的表达水平

与NC组比较，^*P < 0.05。

Figure 3. The expression level of miRNA16 in each group of cells

下载: 全尺寸图片幻灯片

图 4 Bmi-1在各组细胞中的表达水平

与CoC1/DDP组、NC组比较，^*P < 0.05。

Figure 4. The expression level of Bmi-1 in each group of cells

下载: 全尺寸图片幻灯片

图 5 Bmi-1蛋白在各组的条带的IA值水平

Figure 5. IA values of Bmi-1 protein in bands of each group

下载: 全尺寸图片幻灯片

图 6 各组细胞的细胞存活度

与CoC1/DDP组、NC组、过表达质粒组比较，^*P < 0.05。

Figure 6. Cell survival of each group

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图 7 各组细胞的凋亡率水平

与CoC1/DDP组、NC组、过表达质粒组比较，^*P < 0.05。

Figure 7. The apoptosis rate of cells in each group

下载: 全尺寸图片幻灯片

图 8 各组细胞的凋亡率图谱

A：CoC1/DDP细胞组；B：5.0 µM/mL顺铂处理组；C：NC组；D：转染miRNA-15a模拟物组；E：miRNA-16模拟物组；F：过表达的Bmi-1质粒的转染miRNA-15a组；G：过表达的Bmi-1质粒miRNA-16组。

Figure 8. Apoptotic rate atlas of each group

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图 9 各组细胞的γ-H2AX含量

Figure 9. γ-H2AX content in each group

下载: 全尺寸图片幻灯片

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