Epigenetic and genetic analysis of p16 in dermal fibroblasts from type 1 diabetic patients with nephropathy.
Epigenetic and genetic analysis of p16 in dermal fibroblasts from type 1 diabetic patients with nephropathy.
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1 型糖尿病肾病患者真皮成纤维细胞中 p16 的表观遗传学和遗传分析。
DOI：
10.1046/j.1523-1755.2003.00013.x
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Danesh,FarhadR
中科院分区：
文献类型：
--
作者：
Zeng,Lixia;Kanwar,YashpalS;Amro,Nail;Phillips,Carrie;Molitch,Mark;Batlle,Daniel;Danesh,FarhadR
Epigenetic and genetic analysis of p16 in dermal fibroblasts from type 1 diabetic patients with nephropathy.BackgroundSeveral studies have shown that cultured skin fibroblasts from patients with diabetic nephropathy (DN) exhibit a hyperplastic growth phenotype. Increased DNA synthesis in cells from patients with DN may ultimately involve alterations in cell cycle regulatory proteins. p16 protein is a member of INK4 family of cyclin-dependent kinase inhibitors, which plays an important role in cell cycle regulation. In this study, we examined the correlation between p16 protein expression in cultured dermal fibroblasts from type 1 diabetic patients and the presence of DN.MethodWestern blot analysis was performed to compare p16 protein expression in skin fibroblasts from patients with DN as compared to control subjects, diabetic patients without DN, and nondiabetic patients with nephropathy. Transcriptional regulation of thep16gene was assessed using competitive reverse transcription-polymerase chain reaction (RT-PCR). Methylation status of the promoter region of p16 was examined using methylation-specific PCR, and we used single-stranded conformational polymorphism (SSCP)-PCR to assess p16 single-nucleotide polymorphism.ResultsCells from diabetic patients with DN had nondetectable to significantly lower protein expression of p16. Similarly, mRNA expression of p16 was significantly lower in diabetic patients with DN. No hypermethylation ofp16gene was detected, and no abnormal migrating bands were noticed on SSCP-PCR analysis in cells from patients with DN.ConclusionOur data indicate that cells from patients with DN exhibit significantly lower protein and mRNA expression of p16. This study could have not only important implications for the understanding of the pathogenesis of DN, but also the absence of p16 may ultimately serve as an early marker for DN.
