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Resistance of SKW6 cell to apoptosis induction with anti-fas antibody upon transduction of a reverse fragment to a cDNA encoding human 6A8 a-mannosidase

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Science China(Life Sciences) 2001年第4期44卷 365-372页

作者：史耕先靳玉兰王壮志崔巍刘音王讯朱立平 1. Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and Peking Union Medical College 100005 Beijing China

The effect of transduction with a reverse fragment to a cDNA encoding human 6A8 α-mannosidase on apoptosis induction of human B cell line SKW6 by anti-fas antibody was tested. Apoptosis-inducer of anti-fas monoclonal antibody was used to induce apoptosis in SKW6 cells. Giemsa's staining, Annexin-V-FLUOS staining and DNA ladder test were used to determine the events of apoptosis. Indirect immunofluorescent staining with anti-fas antibody was performed to detect the surface fas expression. In a time-course test of 12, 24 and 36 h for apoptosis induction by anti-fas antibody, DNA ladder was observed in the wild-type SKW6 cells in a time-dependent fashion. Mock transduction had no effect on DNA ladder production. However, no DNA ladder was detected in the rAAV-antisense 6A8 cDNA-transduced SKW6. Results from Annexin-V-FLUOS staining on anti-fas antibody-treated cells revealed that the staining-positive rate in the rAAV-antisense 6A8 cDNA-transduced SKW6 cells was decreased in comparison to that in the wild-type and the mock-transduced cells. Giemsa's staining observation showed that the number of dying (with apoptotic bodies) and dead cells was reduced in the rAAV-antisense 6A8 cDNA-transduced SKW6 cells in comparison with that in the wild-type and the mock-transduced cells upon anti-fas antibody induction. The transduction did not affect the expression of fas molecular on cell surface. 100% cells in all the groups showed fas expression. The SKW6 cells became resistant to apoptosis induction by anti-fas antibody upon transduction with a reverse fragment to a cDNA encoding human 6A8 α-mannosidase. The transduction did not affect the expression of fas molecule on cells.

关键词： apoptosis antisense 6A8 cDNA 6A8 α-mannosidase anti-fas antibody B cell line SKW6.

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Partial Beclin 1 silencing aggravates doxorubicin-and fasinduced apoptosis in HepG2 cells

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World Journal of Gastroenterology 2006年第18期12卷 2895-2900页

作者： Fanny Daniel Agnès Legrand Dominique Pessayre Nathalie Vadrot Véronique Descatoire Dominique Bernuau Institut National de la Santé et de la Recherche Médicale INSERMU773Centre de Recherche Bichat Beaujon CRB3BP 416F-75018ParisFrance Université paris 7 Denis Diderot site BichatBP 416F-75018ParisFrance

AIM： To investigate the role of Beclin 1 on the susceptibility of HepG2 cells to undergo apoptosis after anti-fas antibody or doxorubicin treatment. METHODS： Beclin 1 silencing was achieved using RNA interference. DNA ploidy, the percentage of apoptotic cells and the mitochondrial membrane potential were assessed by flow cytometry. Levels of Beclin 1, BCI-XL and cytochrome c, and the cleavage of poly （ADP-ribose） polymerase （PARP） were assayed by using Western blots. RESULTS： Beclin 1 expression decreased by 75% 72 h after Beclin 1 siRNA transfection. Partial Beclin 1 silencing significantly increased the percentage of subG1 cells 24 and 40 h after treatment with doxorubicin or anti-fas antibody, respectively, and this potentiation was abrogated by treatment with a pan-caspase inhibitor. Partial Beclin 1 silencing also increased PARP cleavage, mitochondrial membrane depolarization and cytosolic cytochrome c. The pro-apoptotic consequences of partial Beclin 1 silencing were not associated with a decline in Bcl-XL expression.CONCLUSION： Partial Beclin 1 silencing aggravates mitochondrial permeabilization and apoptosis in HepG2 cells treated with an anti-fas antibody or with doxorubicin.

关键词： Beclin 1 Apoptosis HepG2 cells anti-fas antibody Doxorubicin

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