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BCL2L12A localizes to the cell nucleus and induces growth inhibition through G2/M arrest in CHO cells

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Abstract

BCL2L12, a newly identified member of Bcl-2 family, and its transcript variant BCL2L12A have been found to be associated with favorable prognosis in breast cancer patients while correlated with tumorigenesis of glioblastoma and colon cancer. However, the biological functions of BCL2L12 and especially those of BCL2L12A are largely unknown. Here, we report that, unlike other Bcl-2 family proteins, BCL2L12 and its transcript variant BCL2L12A are nuclear proteins. Interestingly, BCL2L12 forms speckle patterns in the nuclei and potently induces apoptosis in CHO cells. BCL2L12A had a diffuse distribution in the nuclei and inhibits cell growth by inducing cell cycle arrested at G2/M transition in CHO cells. More importantly, BCL2L12A-induced G2/M arrest was associated with a slight up-regulation of cyclin B1 and significant down-regulation of an active form of cyclin B1 phosphorylated at Ser147. Taken together, our study suggests that both BCL2L12 and BCL2L12A have negative effects on CHO cell growths, and that BCL2L12A is a potential cell cycle regulator that interferes with G2–M transition.

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Acknowledgments

This work was supported by National Natural Scientific Foundation of China (30770426), Shanghai Leading Academic Discipline Project (B110), National Science Foundation for Fostering Talents in Basic Research of the National Natural Science Foundation of China (Grant No.J0730860), and State Key Project Specialized for Infectious Diseases (2008ZX10002-015, 2008ZX10002-021, 2008ZX10001-02).

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Authors and Affiliations

  1. Gene Research Center, Shanghai Medical College, Fudan University, 200032, Shanghai, People’s Republic of China

    Yi Hong, Junwu Yang, Yayun Chi, Wenzong Wang, Weibing Wu, Xiaojing Yun, Xiangfei Kong & Jianxin Gu

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  1. Yi Hong
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Correspondence to Jianxin Gu.

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Yi Hong and Junwu Yang have contributed equally to this work.

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11010_2009_233_MOESM1_ESM.jpg

Supplement Fig. 1 Ectopic expression of BCL2L12 induces apoptosis in CHO cells. EGFP or EGFP-tagged BCL2L12 were transiently transfected into CHO cells. (A) 48 hours after transfection, cells were lysed and subjected to Western blot analysis for indicated proteins. (B) All the adherent and non-adherent cells were harvested and subject to FACS analysis as described in “Materials and Methods”. (C) Statistical analysis of cell apoptotic rate from three independent experiments (JPG 105 kb)

11010_2009_233_MOESM2_ESM.jpg

Supplement Fig. 2 Knockdown of BCL2L12 and BCL2L12A did not affect cell cycle progression in MDA-MB-231 cells. (A) Cell cycle distributions of MDA-MB-231/pSilencer and MDA-MB-231/pSi-BCL2L12 cells. (B) Statistical analysis of cell cycle distributions of MDA-MB-231/pSi and MDA-MB-231/pSi-BCL2L12 cells. The data shown are means ± SD of three independent experiments. (C) Lysates of MDA-MB-231/pSilencer and MDA-MB-231/pSi-BCL2L12 cells were subjected to Western blot analysis with antibodies against cell cycle associated proteins (JPG 124 kb)

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Hong, Y., Yang, J., Chi, Y. et al. BCL2L12A localizes to the cell nucleus and induces growth inhibition through G2/M arrest in CHO cells. Mol Cell Biochem 333, 323–330 (2010). https://doi.org/10.1007/s11010-009-0233-z

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