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RNA Pull-down Procedure to Identify RNA Targets of a Long Non-coding RNA
Published on: April 10, 2018
Long Non-Coding RNA MALAT1 Promotes Acute Cerebral Infarction Through miRNAs-Mediated hs-CRP Regulation
1Department of Geriatric Medicine, Shanghai East Hospital, Tongji University School of Medicine, No. 150 Jimo Road, Pudong New Area, Shanghai, 200120, China. tengli0812@126.com.
Abstract:
The occurrence of cerebral infarction commonly takes atherosclerosis as the pathophysiological basis, accompanied by chronic inflammation. Hypersensitive C-reactive protein (hs-CRP) is an important inflammatory factor involved in the formation of atherosclerosis. This study aims to investigate the regulation of hs-CRP expression by long-chain non-coding RNA (LncRNA) MALAT1 in acute cerebral infarction patients. Plasma levels of LncRNA MALAT1 and hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p in 256 Chinese Han ACI patients and 256 controls were analyzed. HUVECs were transfected with LncRNA MALAT1, MALAT1 NC, and si-MALAT1, respectively. The expression levels of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p were analyzed. Then, HUVECs were transfected with hsa-miR-145-5p inhibitor, hsa-miR-140-5p inhibitor, hsa-miR-483-3p inhibitor, hsa-miR-338-3p inhibitor, and hsa-miR-145-5p mimic, hsa-miR-140-5p mimic, hsa-miR-483-3p mimic, hsa-miR-338-3p mimic, and the expression level of hs-CRP was detected by Western blotting. The levels of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p in the plasma of ACI patients were significantly lower than those in the control group (p < 0.001), and the plasma LncRNA MALAT1 levels were significantly higher in ACI patients than in the control group (p < 0.001). The level of LncRNA MALAT1 in plasma of ACI patients and control group was negatively correlated with hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p (r = - 0.36, - 0.79, - 0.76, - 0.75; - 0.60, - 0.68, - 0.48, - 0.56). Plasma levels of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p were negatively correlated with hs-CRP levels in patients with ACI and controls (r = - 0.74, - 0.81, - 0.84, - 0.56; - 0.61, - 0.69, - 0.69, - 0.50). MALAT1 transfection resulted in the decreased levels of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p in HUVECs while overexpression of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p led to a decrease in hs-CRP levels in HUVECs. LncRNA MALAT1 induced the upregulation of CRP expression through inhibiting the expression of hsa-miR-145-5p, hsa-miR-140-5p, hsa-miR-483-3p, and hsa-miR-338-3p.
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