| 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 |
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| 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） |
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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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