The study of the expressions of apoptosis and its related protein by knockdown of human immunodeficiency virus-1 vpr gene / 中华传染病杂志

Objective To observe the expression of apoptosis related protein c-inhibitor of apoptosis protein (IAP)2 by RNA interference of human immunodeficiency virus-1 (HIV-1)vpr gene and analyze the apoptosis of Jurkat cells.Methods Vector (NC),HIV-1vpr (vpr),vpr+ pRNAT-U6.1/Neo-vpr-56 (Si56) and vpr+ pRNAT-U6.1/Neo-vpr-160 (Si160) were transfected to Jurkat cells and cultured for 48 hours.The total RNA and protein were extracted.Expression of vpr gene was detected by reverse transcription (RT)-polymerase chain reaction (PCR) to confirm the success of transfection.Expression of c IAP2 was detected by RT-PCR and Western Blot.The apoptosis of Jurkat cells was observed by flow cytometry.Results Expression of vpr gene was detected in vpr,Si56 and Si160 groups.The mRNA expression levels in Si56 and Si160 groups were significantly lower than that in vpr group,which declined 87.2％ and 82.2％,respectively (P＜0.05).The levels of c IAP2 mRNA expression in vpr,Si56 and Si160 groups were increased by 3.75,2.49 and 2.65 folds,respectively,compared to that in NC group.However,the c-IAP2 mRNA expressions in Si56 and Si160 groups were lower than that in vpr group,which declined 33.7％ and 29.5％,respectively (P ＜ 0.05).The c-IAP2 protein expression was consistent with mRNA by immunoblotting,and those in Si56 and Si160 groups were declined 42.2％ and 46.8％,respectively,compared to that in vpr group (P＜0.05).The apoptosis of J urkat cells was detected in all groups.Compared to NC group,the apoptotic rate in vpr group was increased by 1.76 folds.However,the differences of apoptosis rate among NC,Si56 and Si160 groups were not statistically significant (all P＞0.05).Compared to vpr group,the apoptotic rates in Si56 and Si160 were significantly decreased by 19.26％ and 18.05％,respectively (P＜0.05).Conclusions The expression of c-IAP2 could be downregulated by knockdown of HIV-1 vpr gene in transcription and protein levels,and the apoptosis of Jurkat cells is inhibited.

Identification of a Variant Form of Cellular Inhibitor of Apoptosis Protein (c-IAP2) That Contains a Disrupted Ring Domain

Among the members of the inhibitor of apoptosis (IAP) protein family, only Livin and survivin have been reported to have variant forms. We have found a variant form of c-IAP2 through the interaction with the X protein of HBV using the yeast two-hybrid system. In contrast to the wild-type c-IAP2, the variant form has two stretches of sequence in the RING domain that are repeated in the C-terminus that would disrupt the RING domain. We demonstrate that the variant form has an inhibitory effect on TNF-mediated NF-kappaB activation unlike the wild-type c-IAP2, which increases TNF- mediated NF-kappaB activation. These results suggest that this variant form has different activities from the wild-type and the RING domain may be involved in the regulation of TNF-induced NF-kappaB activation.

Identification of a Variant Form of Cellular Inhibitor of Apoptosis Protein (c-IAP2) That Contains a Disrupted Ring Domain

Among the members of the inhibitor of apoptosis (IAP) protein family, only Livin and survivin have been reported to have variant forms. We have found a variant form of c-IAP2 through the interaction with the X protein of HBV using the yeast two-hybrid system. In contrast to the wild-type c-IAP2, the variant form has two stretches of sequence in the RING domain that are repeated in the C-terminus that would disrupt the RING domain. We demonstrate that the variant form has an inhibitory effect on TNF-mediated NF-kappaB activation unlike the wild-type c-IAP2, which increases TNF- mediated NF-kappaB activation. These results suggest that this variant form has different activities from the wild-type and the RING domain may be involved in the regulation of TNF-induced NF-kappaB activation.

Expression Analysis of c-IAP2 in Ovarian Carcinomas / 대한산부인과학회잡지

OBJECTIVE: Apoptosis or programmed cell death is a normally physiological cell suicide program that is highly conserved among all animals. We previously evaluated overexpression of c-IAP1(Inhibitor of Apoptosis Protein) in ovarian carcinomas compared with normal ovaries. In this study, we demonstrate evidence for the involvement of c-IAP2 in ovarian carcinomas. METHODS: Fresh 9 normal ovaries, 5 benign ovarian cysts and 13 ovarian carcinomas were obtained from routine gynecologic surgeries carried out for benign and malignant ovarian tumors. They were examined for the presence of c-IAP2 by RT-PCR(Reverse Transcriptase Polymerase Chain Reaction), Western blot analysis and immunohistochemical stains. RESULTS: Nine of 14 normal and benign ovarian tumors were negative and 11 of 13 ova rian carcinomas were positive for c-IAP2 by RT-PCR. Positive RT-PCR for c-IAP2 was seen in 11/13 of ovarian carcinomas, a significantly higher percentage than in normal and benign ovarian tumors(5/14). All of these tumors showed strong positive for c-IAP2 by western blot and immunohistochemical staining. Whereas negative RT-PCR for c-IAP2 was seen in 9/14 of normal and benign ovarian tumors, a significantly higher percentage than ovarian carcinomas(2/13). Of these 9 negative samples, 6 had positive Western blot and immunohistochemical stains. There was weak concordance of the result. But expression of c-IAP2 in normal ovarian tissue was localized exclusively in the corpus luteum. Therefore, c-IAP2 may play important role in determining the fate of the follicular destiny. There was no expression in normal ovarian stroma cells for c-IAP2. CONCLUSIONS: These findings suggest that c-IAP2 is expressed in ovarian carcinomas and emerging role in cancer. The c-IAP2 expression has been investigated in the normal ovary, where apoptosis is thought to play an important role in ovulation.

Expression and significance of c-IAP2 and GAS1 in Hodgkin's lymphoma and anaplastic large cell lymphoma / 中国病理生理杂志

AIM: To detect the expression of cytoplasmic inhibitor of apoptosis protein 2 (c-IAP2) and growth arrest-specific gene 1 (GAS1) in Hodgkin's lymphoma (HL) and anaplastic large cell lymphoma (ALCL) and to investigate the role of two genes in the pathogenesis of HL and ALCL.METHODS: HE staining,the antibodies CD30,CD15,CD45RO and CD20 were used to screen the cases of HL and ALCL from 288 cases of lymphoma. The clarified HL and ALCL were subjected for immunohistochemical staining by SP and ABC methods to analyze the expression of c-IAP2 and GAS1. RESULTS: ①The positive rate of c-IAP2 in HL was 25/26(96.1%) while that in ALCL was 6/19(31.6%),there presented statistic significance between HL and ALCL groups( P