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Different gene expression profiles of AD293 and HEK293 cell lines that show contrasting susceptibility to apoptosis induced by overexpression of Bim L

100%

Zhang J., Chen J., Liu L., Ji C., Gu S., Ying K., Mao Y.

2006

vol. 53

issue 3

525-530

Bim is a pro-apoptotic member of the Bcl-2 protein family. Overexpression of Bim proved to be highly cytotoxic for diverse cells.The AD293 cell line is derived directly from the HEK293 cell line but has been transfected with a gene that can improve cell adherence.We found that there was almost no apoptosis seen in Bim L-transfected AD293 cells, but more than half ofBim L-transfected HEK293 cells underwent apoptosis. Suppression subtractive hybridizationwas used to detect the different gene expression profile between these two cell lines. In 192 sequencedpositive clones, there were 30 clones repeating twice or more. Ten genes were selected for identification by semi-quantitative RT-PCR.Thetranscripts of two adhesion-relatedgenes (actin and parvin)and two apoptosis-related genes (cyclin 2 and protein phosphatase 1G) were up-regulated in AD293 cells. These results suggest that the high expression of cell adhesion-related proteins might be responsible for the different apoptosis status after the transfection of Bim L.Our data provide candidate genes responsible for the different apoptosis sensitivity of these two cell lines. Further investigation on thedifferential expression profile between AD293 and HEK293 might improve our understanding of cell apoptosis mechanism.

Overexpression of BimSs3, the novel isoform of Bim, can trigger cell apoptosis by inducing cytochrome c release from mitochondria

84%

Liu L., Chen J., Zhang J., Ji C., Zhang X., Gu S., Xie Y., Mao Y.

2007

vol. 54

issue 3

603-610

Bim is defined as the pro-apoptotic BH3-only protein of the Bcl-2 family, which is a critical sensor and mediator in the mitochondrial-dependent apoptosis. In a previous work, we have cloned a novel transcript of Bim (GenBank accession number: AY305716) from the fetal brain cDNA, which is widely expressed in some carcinoma tissues and normal human tissues. According to the sequence analysis and the newly-defined nomenclature system of Bim isoforms (Adachi et al., 2005, Cell Death Differ 2: 192), we term it BimSs3 according to its characteristic structure. The subcellular location analysis indicated that the fused protein GFP-BimSs3 is distributed in the whole cell, mainly to the nucleus. Overexpression of BimSs3 in HEK293 cells causes apoptosis (28.16 ± 1.55%) compared to the negative control (5.44 ± 2.63%). It also causes cytochrome c release from the mitochondrial fraction to the cytosolic fraction during apoptosis. Western blotting assay indicates the molecular mass of GFP-BimSs3 is approximately 31.0 kDa (GFP: 27 kDa). Hence the open reading frame of BimSs3 may initiate at the second ATG and encodes a 36 amino-acid peptide with BH3 domain.

cDNA cloning, gene organization and expression analysis of human peptidylarginine deiminase type VI.

68%

Zhang J., Dai J., Zhao E., Lin Y., Zeng L., Chen J., Zheng H., Wang Y., Li X., Ying K., Xie Y., Mao Y.

vol. 51

issue 4

1051-1058

Peptidylarginine deiminase (PAD) catalyzes the post-translational modification of protein through the conversion of arginine to citrulline in the presence of calcium ions. Human, similar to rodents, has four isoforms of PAD (type I, II, III and IV/V), each of which is distinct in substrate specificity and tissue specific expression. In our large-scale sequencing project, we identified a new human PAD cDNA from a human fetal brain cDNA library. The putative protein encoded by this cDNA is designated hPADVI. Expression analysis of hPADVI showed that it is mainly expressed in adult human ovary and peripheral blood leukocytes. We conclude that hPADVI may be orthologous to mouse ePAD, basing on sequence comparison, chromosome localization and exon-intron structure analysis. PAD-mediated deimination of epithelial cell keratin resulting in cytoskeletal remodeling suggests a possible role for hPADVI in cytoskeletal reorganization in the egg and in early embryo development. This study describes a new important member of the human PAD family.

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3 Acta Biochimica Polonica

1 2007

1 2006

1 2004

3 Chen J.

3 Mao Y.

3 Zhang J.

2 Gu S.

2 Ji C.

2 Liu L.

2 Xie Y.

2 Ying K.

1 Dai J.

1 Li X.

1 Lin Y.

1 Wang Y.

1 Zeng L.

1 Zhang X.

1 Zhao E.

1 Zheng H.

Search results

Different gene expression profiles of AD293 and HEK293 cell lines that show contrasting susceptibility to apoptosis induced by overexpression of Bim L

Overexpression of BimSs3, the novel isoform of Bim, can trigger cell apoptosis by inducing cytochrome c release from mitochondria

cDNA cloning, gene organization and expression analysis of human peptidylarginine deiminase type VI.