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G-5555 synergized miR-485-5p to alleviate cisplatin resistance in ovarian cancer cells via Pi3k/Akt signaling pathway.
Journal of Reproductive Immunology ( IF 2.9 ) Pub Date : 2020-04-14 , DOI: 10.1016/j.jri.2020.103129
Hai-Feng Qiao 1 , Ying-Lei Liu 1 , Jun You 1 , Yan-Li Zheng 1 , Li-Ping Chen 1 , Xiao-Yan Lu 1 , Lei Du 2 , Feng Shan 1 , Man-Hua Liu 3
Affiliation  

The present study was meant for the discovery of the underlying functions of miR-485-5p in ovarian cancer concerning cisplatin resistance in vitro. RT-qPCR assessed the miR-485-5p expression in ovarian cancer cell lines, normal cells and cisplatin-resistant Cell line OVCA433-CR. After OVCA433-CR treated with 0,3,5umol/L cisplatin, miR-485-5p expressions were determined. MTT observed the cell cytotoxicity in OVCA433-CR after regulation of miR-485-5p. Targets can predicted the putative binding between miR-485-5p and PAK1 and Luciferase Assay verified this. RT-qPCR decided the inhibitory effect in between. MTT tested the cytotoxicity in different combinations of miR-485-5p and PAK1. Western Blot tested the phosphorylation of Pi3k and Akt in response to miR-485-5p and PAK1 interplay. We evaluated the role of Pi3k/Akt signaling in regulation of miR-485-5p and cisplatin resistance with Wortmannin. miR-485-5p was lower expressed in ovarian cancer cells than normal ones and even lower in OVCA433-CR than OVCA433. As the cisplatin concerntration increased, miR-485-5p decreased. miR-485-5p mimics induced lower cisplatin resistance while miR-485-5p inhibitor caused higher resistance. PAK1 targeted miR-485-5p and inhibited miR-485-5p. PAK1 inhibitor helped to lower the resistance to cisplatin caused by miR-485-5p upregulation. miR-485-5p mimics silenced Pi3k/Akt signaling and PAK1 inhibitor aggravated the silencing. Inhibition of Pi3k/Akt signaling increased miR-485-5p, thereby decreasing the cisplatin-resistance in OVCA433-CR. miR-485-5p decreased cisplatin resistance in ovarian cancer cells via Pi3k/Akt signaling, suggesting that miR-485-5p upregulation might alleviate the cisplatin resistance in ovarian patients.

中文翻译:

G-5555 通过 Pi3k/Akt 信号通路协同 miR-485-5p 减轻卵巢癌细胞的顺铂耐药。

本研究旨在发现 miR-485-5p 在卵巢癌中与体外顺铂耐药有关的潜在功能。RT-qPCR 评估了 miR-485-5p 在卵巢癌细胞系、正常细胞和顺铂耐药细胞系 OVCA433-CR 中的表达。OVCA433-CR经0,3,5umol/L顺铂处理后,测定miR-485-5p表达。MTT 在 miR-485-5p 调节后观察到 OVCA433-CR 的细胞毒性。靶标可以预测 miR-485-5p 和 PAK1 之间的推定结合,荧光素酶测定证实了这一点。RT-qPCR 决定了两者之间的抑制作用。MTT 测试了 miR-485-5p 和 PAK1 不同组合的细胞毒性。Western Blot 测试了 Pi3k 和 Akt 响应 miR-485-5p 和 PAK1 相互作用的磷酸化。我们评估了 Pi3k/Akt 信号在调节 miR-485-5p 和 Wortmannin 顺铂耐药中的作用。miR-485-5p 在卵巢癌细胞中的表达低于正常细胞,在 OVCA433-CR 中的表达甚至低于 OVCA433。随着顺铂浓度的增加,miR-485-5p 减少。miR-485-5p 模拟物诱导较低的顺铂耐药性,而 miR-485-5p 抑制剂导致更高的耐药性。PAK1 靶向 miR-485-5p 并抑制 miR-485-5p。PAK1 抑制剂有助于降低由 miR-485-5p 上调引起的对顺铂的耐药性。miR-485-5p 模拟沉默的 Pi3k/Akt 信号,而 PAK1 抑制剂加剧了沉默。Pi3k/Akt 信号通路的抑制增加了 miR-485-5p,从而降低了 OVCA433-CR 中的顺铂耐药性。miR-485-5p 通过 Pi3k/Akt 信号传导降低卵巢癌细胞的顺铂耐药性,
更新日期:2020-04-14
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