Analysis Of Gene Expression Profile In SUDHL6 Cells Of siRNA-Mediated BCL11A Downregulation

The B-cell chronic lymphocytic leukemia (CLL)/lymphoma 11A gene (BCL11A) was associated with hematopoietic malignancies. However, the precise function of this transcription factor in B-cell malignancies still remain poorly characterized. Previous work from our laboratory has shown that BCL11A gene by small interfering RNA (siRNA) resulted in the growth inhibition and apoptosis of the B cell lymphoma cell line (i.e., SUDHL6 and EB1). The aim of this study was to further elucidate the molecular mechanism of this process by analyzing the gene expression profile in SUDHL6 cells after BCL11A knockdown.

FBCL11A siRNA was transfected into SUDHL6 cells to knock down BCL11A expression. Cells were collected, and RNA isolated for transcriptional profiling using the Affymetrix HG-U133 Plus 2.0 array. The global gene expression profile of the BCL11A siRNA-treated SUDHL6 cells was identified and analyzed. Twenty-one differentially expressed genes were further validated and analyzed from the BCL11A siRNA-treated SUDHL6 cells by real-time quantitative reverse transcript-polymerase chain reaction (qRT-PCR).

FThere were 659 genes differentially expressed between the BCL11A siRNA- and negative control-transfected cells. These included 294 upregulated genes and 365 downregulated genes. The differentially expression genes are involved in various signaling pathways including metabolic pathways, focal adhesion, the MAPK signaling pathway, the cell cycle, the JAK-STAT signaling pathway, the TGF-beta signaling pathway, the WNT signaling pathway, apoptosis, and BCR signaling. qRT-PCR validation of the selected differentially expressed genes demonstrated agreement with the microarray analysis. There was a significant difference in the relative expression level of most of the selected genes differentially expressed between the BCL11A siRNA- and negative control siRNA-treated cells (P<0.05). After the transfection of BCL11A siRNA, among the apoptosis-related genes in the BCL-2 family, BCL2L11 was upregulated 7.24-fold, and BCL-2 was downregulated 3.23-fold.

Our results indicate that BCL11A is involved in gene networks with cancer related functions. BCL11A may play a role in gene expression events related to apoptosis.

Li:This work was supported by Grants from National Natural Science Foundation of China (30871091 and 91129720), the Collaborated grant for HK-Macao-TW of Ministry of Science and Technology (2012DFH30060), the Guangdong Science & Technology Project (2012B0506: Research Funding.