The Effect of m6A RNA Methylase METTL14 Regulating YAP1 on Ischemia-reperfusion Injury in HK-2 Cells
LI Lanmei
BAI Weiwei
CHEN Na
SUN Shaoting
WANG Meng
ZHANG Ming
LI Yafen
Abstract：Objective:To explore the effect of N6-methyladenosine(m6A)RNA methyltransferase 14(METTL14)regulating Yes-associated protein 1(YAP1)on ischemia-reperfusion(IR)injury in HK-2 cells.Methods:HK-2 cells were cultured in vitro and divided into the CK group,the IR group,the IR+si-NC group(transfected with si-NC),the IR+si-METTL14 group(transfected with si-METTL14),the IR+si-METTL14+sh-NC group(transfected with si-METTL14+sh-NC),and the IR+si-METTL14+sh-YAP1 group(transfected with si-METTL14+sh-YAP1).Cell viability was detected by thiazole blue tetrazolium bromide(MTT).The levels of lactate dehydrogenase(LDH),tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1β were detected by enzyme-linked immunosorbent assay(ELISA).The kit was used to detect the contents of malondialdehyde(MDA),superoxide dismutase(SOD),and catalase(CAT).Cell apoptosis was detected by flow cytometry.The gene expressions of METTL14 and YAP1 in cells were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR).The protein expressions of Bax,Bcl-2,METTL14,and YAP1 were detected by Western Blot.Results:Compared with the CK group,the expression of METTL14 mRNA in HK-2 cells,the levels of LDH,IL-6,TNF-α,IL-1β in cells,the content of MDA,the apoptosis rate,the m6A modification level of total mRNA in cells,and the protein expressions of Bax and METTL14 in the IR group increased.The mRNA expression of YAP1,cell proliferation rate,activities of SOD and CAT,and the protein expressions of Bcl-2,Ki-67 and YAP1 decreased(P＜0.05).Compared with the IR+si-NC group,the expression of METTL14 mRNA in HK-2 cells,the levels of LDH,IL-6,TNF-α,IL-1β,MDA content,apoptosis rate,the m6A modification level of total mRNA in cells,and the protein expressions of Bax and METTL14 in the IR+si-METTL14 group decreased.The mRNA expression of YAP1,cell proliferation rate,activities of SOD and CAT,and the protein expressions of Bcl-2,Ki-67 and YAP1 increased(P＜0.05).Compared with the IR+si-METTL14+sh-NC group,the levels of LDH,IL-6,TNF-α,IL-1β,MDA content,apoptosis rate,m6A modification level of total mRNA in cells,and Bax expression in HK-2 cells of the IR+si-METTL14+sh-YAP1 group increased.The mRNA expression of YAP1,cell proliferation rate,activities of SOD and CAT,and the protein expressions of Bcl-2,Ki-67 and YAP1 decreased(P＜0.05).Conclusion:Silencing the expression of METTL14 could reduce IR-induced inflammation,oxidative stress and apoptosis of HK-2 cells,and it might be related to increasing YAP1 expression and the reducing m6A modification level.
Keywords：ischemia-reperfusionHK-2 cellsN6-methyladenosine RNA methylasemethyltransferase 14Yes-associated protein 1experimental study
Publication Date：2026-01-25
Online Publishing Date：2026-02-04(First online date of this platform, not the publication date of the document)
Pages：7( 220-226 )