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Archive > Volume 24 Issue 4 > 2017,24(4):373-379. DOI:10.3872/j.issn.1007-385X.2017.04.007 Prev Next
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The effect of RUNX2 on apoptosis of mouse breast cancer 4T1 cells under hypoxia
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Abstract:
Objective:To explore the effect of Runt-related transcription factor 2 (RUNX2) on apoptosis of mouse breast cancer 4T1 cells under hypoxia and its mechanism. Methods: Western blotting and Real-time PCR were used to measure the effect of hypoxia on the expression levels of RUNX1, RUNX2, RUNX3 mRNA and RUNX2 protein in 4T1 cells; small interference RNA (siRNA) and eukaryotic recombinant plasmid DNA over-expression technique were used to down-or up-regulate the RUNX2 expression in 4T1 cells respectively; Co-immunoprecipitation was used to detect the binding of RUNX2 and other proteins in 4T1 cells; Flow cytometry was used for the detection of the effect of down-/up-regulation of RUNX2 on apoptosis of 4T1 cells under normoxic/hypoxic condition. Results: The mRNA expression levels of RUNX1 and RUNX2 in 4T1 cells were up-regulated under hypoxic condition (P<0.05), and the protein expression of RUNX2 was significantly elevated (P<0.05). Transfection with RUNX2-siRNA-1415 or pcDNA3.1(-)-RUNX2 could significantly decrease or increase the RUNX2 level in 4T1 cells, respectively; Under normoxic/hypoxic condition, apoptosis rate of 4T1 cells that transfected with siRNA-RUNX2-1415 was significantly increased by comparing with negative control group(normoxia: \[12.83±0.2404\]% vs \[9.3±0.5508\]%, P<0.05; hypoxia: \[19.77±0.59\]% vs \[15.13±032\]%, P<0.05); however, the apoptosis rate of 4T1 cells were decreased by over-expressing RUNX2 (normoxia: \[9.967±02728\]% vs \[14.07±0.7965\]%, P<0.05; hypoxia: \[22.43±1.02\]% vs \[34.93±0.71\]%, P<005). Under hypoxic condition, the expression levels of hypoxic induced factor-1α (HIF-1α) and RUNX2 were significantly elevated, and RUNX2 could bind with HIF-1α to form complex. Conclusion: Under the hypoxic micro-environment, 4T1 cells express the high level of RUNX2, in the meanwhile, high level RUNX2 could inhibit the apoptosis of tumor cells by interacting with HIF-1α.
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Project Supported:
Project supported by the National High Technology Research and Development Program of China (863 Program) (No.SS2014AA020801)
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