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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

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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.

Isolation and expression pattern of RGS21 gene, a novel RGS member

84%

Li X., Chen L., Ji C., Liu B., Gu J., Xu J., Zou X., Gu S., Mao Y.

2005

vol. 52

issue 4

943-946

Regulators of G-protein signaling (RGS) proteins are known for the RGS domain that is composed of a conserved stretch of 120 amino acids, which binds directly to activated G-protein α subunits and acts as a GTPase-activating protein (GAP), leading to their deactivation and termination of downstream signals. In this study, a novel human RGS cDNA (RGS21), 1795 bp long and encoding a 152-amino acid polypeptide, was isolated by large-scale sequencing analysis of a human fetal brain cDNA library. Unlike other RGS family members, RGS21 gene has no additional domain/motif and may represent the smallest known member of RGS family. It may belong to the B/R4 subfamily, which suggests that it may serve exclusively as a negative regulator of αi/o family members and/or αq/11. PCR analysis showed that RGS21 mRNA was expressed ubiquitously in the 16 tissues examined, implying general physiological roles.

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.

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3 Acta Biochimica Polonica

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3 Gu S.

3 Ji C.

3 Mao Y.

2 Chen J.

2 Liu L.

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1 Gu J.

1 Li X.

1 Liu B.

1 Xie Y.

1 Xu J.

1 Ying K.

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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

Isolation and expression pattern of RGS21 gene, a novel RGS member

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