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miRNA-200b逆转肝细胞癌HepG2细胞对索拉菲尼的耐药性及其相关机制
MiRNA-200b reverses drug resistance of hepatocellular carcinoma HepG2 cells to sorafenib and its underlying mechanism
- 第三军医大学学报 2021年43卷第22期 页码：2435-2440
- 作者机构：
400038 重庆，陆军军医大学（第三军医大学）第一附属医院肝胆外科
- 作者简介：
[ 张雷达，E-mail：2518569931@qq.com ZHANG Leida, E-mail: 2518569931@qq.com
- 基金信息：
DOI：10.16016/j.1000-5404.202105085
中图分类号： R735.7;R915;R979.1收稿：2021-05-20，
修回：2021-08-02，
纸质出版：2021-11-30
- 稿件说明：
移动端阅览
邓永, 唐腾骞, 张雷达. miRNA-200b逆转肝细胞癌HepG2细胞对索拉菲尼的耐药性及其相关机制[J]. 第三军医大学学报, 2021,43(22):2435-2440.
Yong DENG, Tengqian TANG, Leida ZHANG. MiRNA-200b reverses drug resistance of hepatocellular carcinoma HepG2 cells to sorafenib and its underlying mechanism[J]. Journal of Third Military Medical University, 2021, 43(22): 2435-2440.
邓永, 唐腾骞, 张雷达. miRNA-200b逆转肝细胞癌HepG2细胞对索拉菲尼的耐药性及其相关机制[J]. 第三军医大学学报, 2021,43(22):2435-2440. DOI： 10.16016/j.1000-5404.202105085.
Yong DENG, Tengqian TANG, Leida ZHANG. MiRNA-200b reverses drug resistance of hepatocellular carcinoma HepG2 cells to sorafenib and its underlying mechanism[J]. Journal of Third Military Medical University, 2021, 43(22): 2435-2440. DOI： 10.16016/j.1000-5404.202105085.
摘要
目的
2
探究miRNA-200b对肝细胞癌索拉菲尼耐药的影响，以期延缓或改善晚期肝癌患者对索拉菲尼的耐药，增加索拉菲尼的疗效。
方法
2
通过逐步提高培养基中索拉菲尼浓度的方法建立索拉菲尼耐药的肝细胞癌HepG2-SR细胞系，并通过实时荧光定量（RT-PCR）法检测HepG2-SR细胞系和其母本HepG2细胞系中miRNA-200b的表达差异，同时检测HepG2-SR细胞转染miRNA-200b mimics后miRNA-200b表达的变化；CCK-8法检测转染后HepG2-SR细胞对索拉菲尼敏感度的改变；划痕实验和transwell实验检测转染前后HepG2-SR细胞迁移和侵袭能力的改变；流式细胞仪检测转染前后HepG2-SR细胞凋亡情况；Western-blot检测转染前后上皮间质转变（epithelial mesenchymal transition
EMT）标志物的改变。
结果
2
耐药株HepG2-SR细胞对索拉菲尼的耐药性显著高于其母本株HepG2，miRNA-200b的表达显著低于其母本株HepG2；HepG2-SR细胞转染miRNA-200b mimics后miRNA-200b表达显著升高，索拉菲尼半抑制浓度（IC
50
）显著降低，细胞迁移率显著降低，细胞侵袭能力显著降低，细胞凋亡显著增加，上皮细胞标志E-Cadherin表达增加，间质细胞标志N-Cadherin、Vimentin表达减弱。
结论
2
miRNA-200b抑制HepG2-SR细胞的增殖、迁移、侵袭和EMT，促进细胞凋亡，从而逆转索拉菲尼耐药性。
Abstract
Objective
2
To explore the effect of miRNA-200b on sorafenib resistance in hepatocellular carcinoma cells in order to increase the efficacy of the drug in advanced liver cancer patients.
Methods
2
Sorafenib resistant hepatocellular carcinoma HepG2-SR cell line was established by gradually increasing the concentration of sorafenib in the culture medium. The expression of miRNA-200b in HepG2-SR cells
HepG2 cells and miRNA-200b transfected HepG2-SR cells was detected by real-time fluorescence quantitative assay (RT-PCR). In HepG2-SR cells before and after transfection
sorafenib sensitivity was detected by CCK-8 assay
migration and invasion abilities were detected by wound-healing assay and Transwell assay
cell apoptosis was measured by flow cytometry
and expression levels of epithelial mesenchymal transition (EMT) markers were studied by Western blotting.
Results
2
Sorafenib resistance was significantly higher and expression of miRNA-200b was significantly lower in HepG2-SR cells than HepG2 cells. Transfection of miRNA-200b mimics resulted in significantly increased miRNA-200b expression
decreased IC
50
of sorafenib
reduced cell migration rate and cell invasion
enhanced apoptosis and increased expression of E-cadherin but decreased expression of N-cadherin and Vimentin.
Conclusion
2
MiRNA-200b reversed sorafenib resistance of HepG2-SR cells by inducing apoptosis and inhibiting cell proliferation
migration
invasion and EMT.
关键词
Keywords
references
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