刘渤娜,杜 成,郑振东.miR-142-5p靶向DDX5调控卵巢癌顺铂耐药[J].肿瘤学杂志,2022,28(7):539-544.
miR-142-5p靶向DDX5调控卵巢癌顺铂耐药
miR-142-5p Targeting DDX5 in Regulating Cisplatin Resistance in Ovarian Cancer
投稿时间:2021-10-21  
DOI:10.11735/j.issn.1671-170X.2022.07.B002
中文关键词:  卵巢癌  miR-142-5p  DDX5  顺铂  耐药
英文关键词:ovarian cancer  miR-142-5p  DDX5  cisplatin  drug resistance
基金项目:辽宁省科学技术计划项目书(2019-ZD-1050)
作者单位
刘渤娜 中国人民解放军北部战区总医院 
杜 成 中国人民解放军北部战区总医院 
郑振东 中国人民解放军北部战区总医院 
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中文摘要:
      摘 要:[目的] 分析微小RNA(microRNA,miR)-142-5p靶向DEAD box p68 RNA解旋酶(DEAD-box RNA helicase 5,DDX5)调控卵巢癌顺铂(cisplatin,DDP)耐药机制。[方法] 收集2019年4月至2020年11月45例卵巢癌手术切除的癌组织及癌旁组织,采用RT-qPCR测定miR-142-5p表达,Western blot法测定DDX5表达。实验分4组:SKOV3组、SKOV3/DDP组、阴性对照组(感染阴性对照慢病毒LV-miR-142-5p-NC的SKOV3/DDP细胞)、病毒感染组(感染沉默miR-142-5p的慢病毒LV-miR-142-5p-IN的SKOV3/DDP细胞),对比各组miR-142-5p、DDX5表达、细胞生长抑制率、细胞凋亡率和细胞OD值。 双荧光素酶报告实验验证miR-142-5p和DDX5的靶向关系。[结果] 卵巢癌组织miR-142-5p mRNA表达、DDX5蛋白表达比癌旁组织高(P<0.05);SKOV3/DDP组、阴性对照组miR-142-5p mRNA表达和DDX5蛋白表达比SKOV3组高(P<0.05);病毒感染组miR-142-5p mRNA表达、DDX5蛋白表达相比SKOV3/DDP组、阴性对照组低(P<0.05)。 不同浓度顺铂(1.25、2.5、5、10、20 μg/mL)作用后,SKOV3/DDP组、阴性对照组的细胞生长抑制率比SKOV3组低(P<0.05);病毒感染组细胞生长抑制率比SKOV3/DDP组及阴性对照组高(P<0.05)。DDP培养48 h 后,SKOV3/DDP组、阴性对照组细胞凋亡率比SKOV3组低,OD值比SKOV3组高(P<0.05);相比SKOV3/DDP组和阴性对照组,病毒感染组凋亡率高,OD值低(P<0.05)。DDX5 WT+miR-142-5p mimic组荧光素酶活性比DDX5 WT组高(P<0.05)。[结论] miR-142-5p下调可能通过抑制DDX5表达促进细胞凋亡、抑制细胞增殖,从而逆转卵巢癌细胞对顺铂的耐药性。
英文摘要:
      Abstract:[Objective] To analyze the mechanism of microRNA(miR)-142-5p targeting DEAD box P68 RNA helicase(DDX5) in regulating cisplatin resistance in ovarian cancer. [Methods] From April 2019 to November 2020, the cancer tissues and adjacent tissues of 45 cases of ovarian cancer were collected. The expression of miR-142-5p was measured by RT-qPCR and the expression of DDX5 was measured by Western blot. Human ovarian carcinoma cell line and cisplatin resistant cell line SKOV3/DDP were used for in vitro experiments(SKOV3 group and SKOV3/DDP group), the SKOV3/DDP cells were infected with negative control lentivirus LV-miR-142-5p-NC(negative control group) or with lentivirus LV-miR-142-5p-in silencing miR-142-5p(miR-142-5p positive group). The expression of miR-142-5p and DDX5, cell growth inhibition rate, apoptosis rate and cell opitical density(OD) were compared among groups. Double luciferase report assay was used to verify the targeting relationship between miR-142-5p and DDX5. [Results] The expressions of miR-142-5p mRNA and DDX5 protein in ovarian cancer were higher than those in adjacent tissues(P<0.05). The expression of miR-142-5p mRNA, DDX5 protein in SKOV3/DDP group and negative control group were higher than those in SKOV3 group(P<0.05). The expression of miR-142-5p mRNA, DDX5 protein in miR-142-5p positive group were lower than those in SKOV3/ DDP group and negative control group(P<0.05). After treated with DDP(1.25, 2.5, 5, 10 and 20 μg/mL respectively), the cell growth inhibition rate in SKOV3/DDP group and negative control group was lower than that of SKOV3 group(P<0.05). The cell growth inhibition rate of miR-142-5p positive group was higher than that of SKOV3/DDP group and negative control group(P<0.05). The apoptosis rate of SKOV3 / DDP group and negative control group at 48h was lower than that of SKOV3 group, and the OD value was higher than that of SKOV3 group(P<0.05). The apoptosis rate in miR-142-5p positive group was higher than that in SKOV3 / DDP group and negative control group at 48 h, and the OD value was lower than that in SKOV3 / DDP group and negative control group(P<0.05). The luciferase activity of DDX5 WT + miR-142-5p mimic group was higher than that of DDX5 WT group(P<0.05). [Conclusion] The down-regulation of miR-142-5p can promote apoptosis and inhibit cell proliferation by inhibiting the expression of DDX5, to reverse the resistance of ovarian cancer cells to cisplatin.
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