BACKGROUND: The obesity has led to a plenty of diseases including hypertension, coronary heart disease, fatty liver, hyperlipidemia, and type 2 diabetes. Therefore, understanding the mechanism of adipocyte differentiation is of far-reaching significance to the prevention and treatment of obesity. For the current studies of the mechanism of adipocyte differentiation pay more attention to microRNA, rather than long non-coding RNAs (lncRNAs).
OBJECTIVE: To obtain the lncRNAs whose fold change was apparent during adipogenic differentiation, and to further screen the lncRNAs that possibly play a crucial role in adipogenic differentiation for verification.
METHODS: Subcutaneous fat was obtained from human abdomen. Adipose-derived stem cells were collected using tissue culture method. The third passage of adipose-derived stem cells was used for adipogenic differentiation. Through microarray technology, the expression levels of lncRNAs and mRNA were analyzed at 0, 5 and 12 days in adipogenic differentiation. Combining with bioinformatics report, lncRNAs apparently presented fold change were screened and verified by qRT-PCR.
RESULTS AND CONCLUSION: Fold change 1.5 (P < 0.05) was considered as a criterion during adipogenic differentiation. The number of up-regulated lncRNAs was 748 for 5 days versus 0 day, 847 for 12 days versus 0 days, 593 for 12 days versus 5 days. At the same time, the down-regulated number was 828 for 5 days versus 0 day, 1 113 for 12 days versus 0 day, 750 for 12 days versus 5 days during adipogenic differentiation. In
combination with bioinformatics analysis results, 3 of 28 lncRNAs were related to lipid metabolism: AK304548, BP216319 and DA852857, according to the standard that fold change in 0, 5 and 12 days was higher, and the target genes were known to be associated with adipogenesis-related genes. PCR results showed that the expression of AK304548 and BP216319 and its target gene presented an up-down trend, which is consistent with the microarray sequencing results. These results indicated that lncRNA plays a critical regulatory role in the adipogenic differentiation.