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. 2019 Nov;33(9):e22985.
doi: 10.1002/jcla.22985. Epub 2019 Jul 31.

Transcription factor CEBPB inhibits the proliferation of osteosarcoma by regulating downstream target gene CLEC5A

Jianhua Lu  1 , Weikai Chen  1 , Hao Liu  1 , Huilin Yang  1 , Tao Liu  1
Affiliations

Transcription factor CEBPB inhibits the proliferation of osteosarcoma by regulating downstream target gene CLEC5A

Jianhua Lu et al. J Clin Lab Anal. 2019 Nov.

Abstract

Objective: To screen the gene related to osteosarcoma (OS) metastasis and the molecular mechanism.

Methods: GEO database and R2 chip analysis platform were used to screen genes related to OS metastasis. UCSC gene browser was used to find the transcription factor (TF) of CLEC5A. The mRNA level and protein expression of CLEC5A in OS tissues and normal tissues were determined by RT-PCR and Western blotting, respectively. OS cell lines MG-63 were transfected with CEBPB recombinant plasmid. After transfection, the expression of CLEC5A in MG-63 cells was determined and the cell proliferation situation was determined by clone formation assay.

Results: Three genes CLEC5A, ALOX5AP, and RNASE3 were obtained, and CLEC5A has the highest correlation with OS. CLEC5A has screened the gene related to OS metastasis, and CEBPB can be taken as TF regulating downstream gene CLEC5A. CEBPB can regulate the downstream CLEC5A as transcription factor. The relative mRNA level and protein expression of CLEC5A in OS tissues were significantly higher than those in normal tissues. CLEC5A can prevent OS metastasis. Transfection of CEBPB increased the expression of CLEC5A in MG-63 cells and also inhibited the proliferation of OS.

Conclusion: CEBPB can inhibit the proliferation of OS cells via regulating the expression of CLEC5A.

Keywords: C-type lectin domain family 5; enhancer-binding protein beta; member A; osteosarcoma; proliferation; transcription factor.

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Conflict of interest statement

All of the authors have no conflict of interest in this research.

Figures

Figure 1
Figure 1
UCSC gene browser analysis to explore the transcription factors regulating CLEC5A
Figure 2
Figure 2
Effect of CLEC5A on the metastasis survival probability of osteosarcoma
Figure 3
Figure 3
CLEC5A was down‐regulated in osteosarcoma. A, The relative mRNA level of CLEC5A in tumor tissues and the adjacent normal tissues by RT‐PCR. **P < 0.01 vs normal group, indicating the difference has statistical significance. B, The protein expression of CLEC5A in tumor tissue and the adjacent normal tissues by Western blotting assay
Figure 4
Figure 4
CEBPB is the transcription factor of CLEC5A. A, The relative mRNA level of CLEC5A in NC group and CEBPB group by RT‐PCR. **P < 0.01 vs NC group, indicating the difference has statistical significance. B, The protein expression of CLEC5A in NC group and CEBPB group by Western blotting assay
Figure 5
Figure 5
The clone formation situation of osteosarcoma cells MG‐63 in NC and CEBPB group was determined by clone formation assay
See this image and copyright information in PMC

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