miR-499a-5p通过靶向Pten减轻嗜铬细胞瘤细胞PC12低氧损伤

doi:10.16352/j.issn.1001-6325.2022.12.1873

基础医学与临床 ›› 2022, Vol. 42 ›› Issue (12): 1873-1878.doi: 10.16352/j.issn.1001-6325.2022.12.1873

miR-499a-5p通过靶向Pten减轻嗜铬细胞瘤细胞PC12低氧损伤

张丽阳^*, 孙军, 陈迪

南阳市中心医院神经内科, 河南南阳 473000

收稿日期:2021-12-08 修回日期:2022-05-22 出版日期:2022-12-05 发布日期:2022-11-23
通讯作者: ^* zhangliyang855@tom.com
基金资助:
河南省科学技术厅基金(192102310349)

miR-499a-5p alleviates hypoxic damage of pheochromocytoma cells through targeting at Pten

ZHANG Li-yang^*, SUN Jun, CHEN Di

Department of Neurology, Nanyang Central Hospital, Nanyang 473000, China

Received:2021-12-08 Revised:2022-05-22 Online:2022-12-05 Published:2022-11-23
Contact: ^* zhangliyang855@tom.com

摘要/Abstract

摘要： 目的研究miR-499a-5p对化学性低氧诱导的嗜铬细胞瘤细胞(PC12)损伤的影响。方法将PC12细胞分为对照组、低氧组(CoCl₂处理24 h)、低氧+miR-NC组和miR-mimic转染组(分别转染miR-NC和miR-499a-5p mimic后,用CoCl₂处理24 h)。用RT-qPCR检测miR-499a-5p表达;用MTT法检测细胞活性;用试剂盒检测乳酸脱氢酶(LDH)活性;用流式细胞测量术检测各组细胞凋亡;用Western blot检测磷酸酶与张力蛋白同源物(PTEN)蛋白的表达。用荧光素酶报告基因检测Pten是否为miR-499a-5p的直接作用靶标。结果与对照组比较,低氧组中miR-499a-5p表达和细胞活性降低(P＜0.01或P＜0.001),LDH活性和细胞凋亡增加(P＜0.01或P＜0.001);过表达miR-499a-5p可提高低氧条件下PC12细胞的存活率并抑制其凋亡(P＜0.05或P＜0.01)。Pten是miR-499a-5p的直接作用靶标。结论 miR-499a-5p通过靶向Pten减轻PC12细胞低氧损伤。

关键词: miR-499a-5p, 磷酸酶与张力蛋白同源物, PC12细胞, 低氧损伤

Abstract: Objective To study the effect of miR-499a-5p on injury induced by chemical hypoxia of pheochromocytoma cell(PC12). Methods PC12 cells were divided into control group, hypoxia group (treated with CoCl2 for 24 h), hypoxia+miR-NC or miR-mimic groups. Cell viability was detected by MTT assay; the expression of miR-499a-5p was detected by RT-qPCR; lactate dehydrogenase (LDH) activity was detected by kit; apoptosis was detected by flow cytometry; the expression of phosphatase and tensin homolog (PTEN) protein was detected by Western blot. The luciferase reporter gene was used to detect whether Pten was a direct target of miR-499a-5p. Results Compared with control group, the cell viability and miR-499a-5p expression decreased (P＜0.01 or P＜0.001), LDH activity and apoptosis increased (P＜0.01 or P＜0.001) in hypoxia group. Over-expression of miR-499a-5p enhanced survival rate and inhibited apoptosis of PC12 cells under hypoxia (P＜0.05 or P＜0.01). Pten was a direct target of miR-499a-5p. Conclusions miR-499a-5p alleviates hypoxic injury of PC12 cells through targeting at Pten.

Key words: miR-499a-5p, phosphatase and tensin homolog, PC12 cell, hypoxic injury

中图分类号:

R966

张丽阳, 孙军, 陈迪. miR-499a-5p通过靶向Pten减轻嗜铬细胞瘤细胞PC12低氧损伤[J]. 基础医学与临床, 2022, 42(12): 1873-1878.

ZHANG Li-yang, SUN Jun, CHEN Di. miR-499a-5p alleviates hypoxic damage of pheochromocytoma cells through targeting at Pten[J]. Basic & Clinical Medicine, 2022, 42(12): 1873-1878.

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miR-499a-5p通过靶向Pten减轻嗜铬细胞瘤细胞PC12低氧损伤

miR-499a-5p alleviates hypoxic damage of pheochromocytoma cells through targeting at Pten

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