Objective To study the role of CRIF1 in high glucose induced rRNA gene transcription.Methods HEK293T cells were transfected with CRIF1 ectopic expression vector or control vector for 48 hours were collected.Total RNA was extracted to detect the mRNA level of pre-rRNA.HEK293T cells were cultured in DMEM medium containing 0,1,4.5g/L glucose for 12hours.The whole cell lysate were collected and CRIF1 was detected by Western blot assay.HEK293T cells were transfected with shCRIF1 vector or control vector for 36 hours and cultured in DMEM medium containing 0,1,4.5g/L glucose for another 12 hours were collected.Total RNA was extracted to detect the mRNA level of pre-rRNA.Results CRIF1 promoted the expression of pre-rRNA.CRIF1 expression gradually elevated with the increasing concentration of glucose.Knocking down of CRIF1 repressed the high expression of pre-rRNA depended on glucose.Conclusion CRIF1 promotes the pre-rRNA expression induced by glucose and plays an important role in glucose-induced rRNA gene transcriptional regulation.%目的 探索CRIF1在高糖诱导下对rRNA基因转录中的作用.方法 在HEK293T细胞中,转染CRIF1表达载体或空载对照,培养48h后收取细胞提取总RNA,通过RT-qPCR检测pre-rRNA的mRNA水平;用含0、1、4.5g/L糖浓度的培养基培养HEK293T细胞12h后,收集全细胞提取物,进行Western blot法检测CRIF1蛋白的表达水平;在HEK293T细胞中转染shCRIF1干扰质粒或空载对照,36h后换用含0、1、4.5g/L糖浓度的培养基继续培养细胞12h后,收取细胞提取总RNA,通过RT-qPCR检测pre-rRNA的水平.结果 CRIF1可促进pre-rRNA的表达;随着糖浓度的升高,CRIF1的表达量逐渐增加;敲低CRIF1可抑制糖浓度依赖的pre-rRNA的表达.结论 CRIF1可提高高糖诱导下pre-rRNA的表达水平,从而参与高糖诱导的rRNA基因的转录调控.
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