| 缪玖麟,王建荣,徐志敏,等.微小 RNA?25?3p靶向下调甘油磷酸二酯酶磷酸结构域 5基因对乳腺癌细胞顺铂耐药性的影响[J].安徽医药,2020,24(10):1937-1942. |
| 微小 RNA?25?3p靶向下调甘油磷酸二酯酶磷酸结构域 5基因对乳腺癌细胞顺铂耐药性的影响 |
| Effect of miR?25?3p targeted down?regulation of glycerophosphodiester phosphodiesterase domain 5 gene on cisplatin resistance of breast cancer cells |
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| DOI：10.3969/j.issn.1009?6469.2020.10.006 |
| 中文关键词: 乳腺肿瘤 抗药性，肿瘤 miR?25?3p 甘油磷酸二酯酶磷酸结构域 5 顺铂 谷胱甘肽转移酶 |
| 英文关键词: Breast neoplasms Drug resistance,neoplasm microRNA?25?3p Glycerophosphodiester phosphodiesterase do? main 5 Cisplatin Glutathione transferase |
| 基金项目: |
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| 中文摘要: |
| 目的探讨微小 RNA?25?3p(miR?25?3p)靶向甘油磷酸二酯酶磷酸结构域 5(GDPD5)对乳腺癌细胞顺铂耐药性的影响及分子机制。方法本研究起止时间为 2018年 10月至 2019年 6月，人乳腺癌细胞 MCF?7和乳腺癌顺铂耐药细胞 MCF?7/DDP购于中国科学院上海细胞研究所，采用实时荧光定量 PCR(qRT?PCR)和蛋白质印迹法(Western blot)检测正常乳腺癌细胞 MCF?7和乳腺癌顺铂耐药细胞 MCF?7/DDP中 miR?25?3p、GDPD5和谷胱甘肽巯基转移酶 π(GST-π)的表达水平。四甲基偶氮唑盐比色(MTT)法检测过表达 miR?25?3p或过表达 miR?25?3p联合顺铂对 MCF?7/DDP细胞的增殖抑制作用，蛋白质印迹法检测 GDPD5、GST?π和细胞周期蛋白 D1(Cyclin D1)蛋白的表达水平。双荧光素酶报告基因实验和蛋白质印迹法验证 miR?25?3p和 GDPD5的靶向关系。结果与 MCF?7细胞相比， MCF?7/DDP细胞中 miR?25?3p的表达显著下调， GDPD5和 GST?π的表达显著上调。过表达 miR?25?3p后， MCF?7/DDP细胞存活率显著降低［(50.36±5.04)%比(100.00±10.11)%］，CyclinD1相对表达水平降低(0.40±0.05)比(1.12±0.11)GST?π表达水平降低(0.35±0.04)比(1.15±0.12)；且过表达 miR?25?3p可降低 MCF?7/DDP细胞的顺铂耐药性。 miR?25?3p可靶向，负性调控 GDPD5的表达。过表达 GDPD5可部分逆转 miR?25?3p对 MCF?7/DDP细胞增殖和顺铂耐药性的影响。结论 miR?25?3p通过靶向下调 GDPD5抑制 MCF?7/DDP细胞存活，进而降低 MCF?7/DDP细胞对顺铂的耐药性。 |
| 英文摘要: |
| Objective To investigate the effect of microRNA?25?3p(miR?25?3p)targeting glycerophosphodiester phosphodiester? ase domain 5(GDPD5)on cisplatin resistance in breast cancer cells and its molecular mechanism.Methods The study was con?ducted from October 2018 to June 2019.Human breast cancer cells MCF?7 and cisplatin?resistant breast cancer cells MCF?7/DDPwere purchased from Shanghai Institute of Cell Research，Chinese Academy of Sciences.Real?time fluorescent quantitative PCR(qRT?PCR)and Western blot were used to detect the expression level of miR?25?3p，GDPD5 and glutathione transferase(GST?π) in breast cancer cells MCF?7 and cisplatin?resistant breast cancer MCF?7/DDP cells.Methyl thiazolyl tetrazolium assay(MTT)assaywas used to detect the inhibitory effect of over?expressing miR?25?3p or over?expressing miR?25?3p combined with cisplatin on theproliferation of MCF?7 /DDP cells，and Western blot was used to detect the expression of GDPD5，GST?π and CyclinD1 proteins.The dual luciferase reporter gene assay and Western blot were used to verify the targeting relationship between miR?25?3p andGDPD5.Results Compared with MCF?7 cells，the expression of miR?25?3p in MCF?7/DDP cells was significantly down?regulated， while the expression of GDPD5 and GST?π was significantly up?regulated.After overexpression of miR?25?3p，the survival rate of MCF?7/DDP cells was significantly reduced［(50.36±5.04)% vs.(100.00±10.11)%］and the relative expression of CyclinD1 was decreased(0.40±0.05)vs.(1.12±0.11)； the expression of GST?π wasdecreased(0.35±0.0，4)vs.(1.15±0.12)； and overexpression ofmiR?25?3p could reduce the cisplatin resistance of MCF?7/DDP cells.miR?25?3p negatively regulated the expression of GDPD5.Over?expression of GDPD5 partially reversed the effect of miR?25?3p on the proliferation and cisplatin resistance of MCF?7/DDP cells.Conclusion miR?25?3p inhibits the survival of MCF?7/DDP cells by down?regulating GDPD5，thereby reducing the resistance of MCF?7/DDP cells to cisplatin. |
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