Login to Access Video or Poster Abstract: MP39-07
Sources of Funding: National Natural Science Foundation of China (81670611).

Introduction

Tuberous sclerosis complex (TSC) is a multisystem genetic disorder caused by mutations in the TSC1 and TSC2 genes. Over 80% of TSC patients developed angiomyolipomas (TSC-AML), but the molecular events contributing to TSC-AML in TSC are not well understood. However, little is known about the role of microRNAs in TSC-AML.

Methods

Total RNA was used to analyze miRNA and mRNA expression via miRCURY-TM Hy3-TM/Hy5-TM Power labeling kit and Human 12 x 135K Gene Expression Array, respectively. Both miRNA and gene expression profiles were integrated by correlation analysis to identify dysregulated miRNAs with their corresponding predicted target mRNA. Microarray data were validated with qRT-PCR. Regulation of BCL2 like 11 (BCL2L11) expression by miR-9-5p, miR-124-3p and miR-132-3 was determined by luciferase reporter assays. All analyses used a significance level of 0.05 and were generated in SPSS19.0 software.

Results

Using microarray profiling of 3100 miRNAs and 45033 mRNA transcripts, the analysis indicated that 350 of these miRNAs and 37284 mRNAs were expressed in TSC-AML, of which the relative expression of 16 miRNAs and 2881 mRNAs (fold change ≥ 1.5 or ≤ 0.67) was differentially expressed in TSC-AML as compared to non-TSC-AML. The validated results revealed that miR-9-5p, miR-124-3p and miR-132-3p were upregulated whereas BCL2L11 was downregulated in patients with TSC-AML. Further studies revealed that downregulation of miR-9-5p, miR-124-3p or miR-132-3p promoted TSC2-deficient angiomyolipoma-derived cell apoptosis. Moreover, luciferase results and western blot analysis confirmed that BCL2L11 was a target of miR-9-5p, miR-124-3p and miR-132-3p.

Conclusions

In conclusion, we identified a number of miRNAs that are differentially expressed between TSC-AML and non-TSC-AML and constructed posttranscriptional regulatory network miRNA-target gene pairs: BCL2L11 is an endogenous target of miR-9-5p, miR-124-3p and mir-132-3p in the TSC-AML of patients. Downregulation the expression of miR-9-5p, miR-124-3p and miR-132-3 could inhibit proliferation and promote apoptosis in comparison to negative controls TSC2-deficient AML-derived cell.

Funding

National Natural Science Foundation of China (81670611).