Progress of X-linked inhibitor of apoptosis deficiency in hemophagocytic lymphohistiocytosis / 国际儿科学杂志

Hemophagocytic lymphohistiocytosis(HLH)is a systemic hyperinflammation syndrome.XIAP deficiency is an innate immunodeficiency disorder, often combined with HLH, with increased susceptibility to EB virus, mostly in childhood.XIAP deficiency leads to innate immune deficiency, increased apoptosis, and excessive activation of NLRP3 inflammasome, which together lead to impaired clearance of pathogens and excessive release of cytokines, resulting in HLH.The course of HLH in patients with XIAP deficiency is generally mild and the mortality rate is low, but it frequently relapses.In addition to HLH, XIAP deficiency can be combined with some auto-inflammatory disease.Hematopoietic stem cell transplantation is the only curable method and reduced-intensity conditioning is recommended.Some new targeted therapies, such as IFN-γ monoclonal antibodies, IL-18 and IL-6 antagonists may be effective, but the specific efficacy needs more study.

Diallyl disulfide downregulates XIAP through miR-7 to inhibit proliferation, migration and invasion of human gastric cancer SGC7901 cells / 中国药理学通报

Aim To investigate whether diallyl disul¬fide ( DADS) downregulates X-linked inhibitor of ap- optosis protein ( XIAP) through miR-7 , inhibiting the proliferation, migration anrl invasion of gastric eaneer SGC7901 eel Is.Methods Gastric eaneer SGC7901 eell line overexpressing XIAP was established.qRT- PCR and Western blot were used to deteet the effeet of DADS on XIAP expression in overexpressed eells.CCK-8 and plate elone formation assay were used to analyze the effeet of DADS on the proliferation of XIAP overexpressing eells.Wound healing and Transwell ex¬periments were employed to analyze the effeets of DADS on the migration and invasion of XIAP overex¬pressing eells.qRT-PCR was used to deteet the effeet of DADS on the expression of miR-7.Negative regula¬ tion of miR-7 on XIAP was verified by using dual lucif- erase reporter gene assay, qRT-PCR ,anrl Western blot.Results XIAP overexpression enhanced the prolifera¬tion , migration and invasion of SGC7901 cells.DADS downregulated XIAP and inhibited the proliferation, mi¬gration and invasion of XIAP overexpressing cells.DADS upregulated miR-7 expression.miR-7 bound and regulated XIAP 3 'UTR directly.Overexpression of miR-7 decreased XIAP expression,while knockdown of miR-7 increased XIAP expression.Conclusion DADS downregulates XIAP through miR-7 to inhibit SGC7901 cell proliferation, migration and invasion.

The Effect of XIAP and Its Application in Tumor / 中国生物化学与分子生物学报

It has been found that X-linked inhibitor of apoptosis (XIAP) is the most characteristic and strongest inhibition of apoptosis proteins. The characteristic structures of XIAP are BIR domain and RING domain, which are important structures for XIAP to play an anti- apoptotic role. A variety of endogenous inhibitory proteins (XAF1, SMAC and OMI) can also inhibit XIAP’s anti-apoptotic effect in different ways. XIAP can directly inhibit the initiation and persistence of apoptosis pathway of caspase. XIAP participates in the death receptor pathway and mitochondrial pathway of inhibiting apoptosis in a variety of ways, such as directly binds to caspases, and activates NF-кB way and other signal pathways, which is essential for regulating the survival and development of tumor cells. XIAP is highly expressed in many kinds of tumor tissues. The high expression of XIAP is closely related to the occurrence and development, drug resistance, treatment and prognosis of tumors. XIAP deletion can significantly reduce the tumorigenicity of tumor cells, and XIAP is the downstream factor of cell apoptosis formed by blocking multiple signal pathways. Therefore, XIAP has become a new target for clinical anticancer drug design. At present, three potential directions of XIAP application in clinical treatment of cancer are small molecule inhibitors, antisense oligonucleotide inhibitors and XIAP gene silencing. This paper will introduce the biological function of XIAP against apoptosis and its role in the occurrence and development, drug resistance, treatment and prognosis of tumor diseases.

Efficacy of phosphocreatine pre-administration on XIAP and Smac in ischemic penumbra of rats with focal cerebral ischemia reperfusion injury

Abstract Purpose: To observe the efficacy of phosphocreatine pre-administration (PCr-PA) on X-linked inhibitor of apoptosis protein (XIAP), the second mitochondia-derived activator of caspase (Smac) and apoptosis in the ischemic penumbra of rats with focal cerebral ischemia-reperfusion injury (CIRI). Methods: A total of 60 healthy male Sprague Dawley (SD) rats were randomly divided into three groups (n=20): group A (the sham operation group), group B <intraperitoneally injected with 20 mg/kg (10 mg/ml) of saline before preparing the ischemia-reperfusion (IR) model>, and group C <intraperitoneally injected with 20 mg/kg (10 mg/ml) of PCr immediately before preparing the IR model>. After 24 h for reperfusion, the neurological function was evaluated and the tissue was sampled to detect expression of XIAP, Smac and caspase-3 positive cells in the ischemic penumbra so as to observe the apoptosis. Results: Compared with group B, neurological deficit scores, numbers of apoptotic cells, expression of Smac,caspase-9 and the numbers of Caspase-3 positive cells were decreased while expression of XIAP were increased in the ischemic penumbra of group C. Conclusions: Phosphocreatine pre-administration may elicit neuroprotective effects in the brain by increasing expression of X-linked inhibitor of apoptosis protein, reducing expression of second mitochondia-derived activator of caspase, and inhibiting the apoptosis in the ischemic penumbra.

X-linked inhibitor of apoptosis deficiency manifested as Crohn's disease: a case report and literature review / 中华儿科杂志

Objective@#To analyze the clinical characteristics of X-linked inhibitor of apoptosis (XIAP) deficient patients with clinical manifestation of Crohn's disease.@*Methods@#Clinical manifestations, laboratory investigations, genetic testing and therapeutic interventions of one case of XIAP deficiency who was admitted to Department of Gastroenterology in Children's Hospital, Zhejiang University School of Medicine in May 2016 were summarized. PubMed and Chinese database for articles published from January 2016 to June 2017 were searched using the key words of'Crohn's disease’and'XIAP’, and the relevant literature was reviewed.@*Results@#The case we reported was a 6-year-1-month-old boy with recurrent bloody stool for 2 months, and abdominal pain with fever for 2 weeks. The patient had a past history of hemophagocytic lymphohistiocytosis (HLH) and epilepsy in the past one year. Complete blood cell count showed mild anemia (Hb108 g/L). The patient had an elevated high-sensitivity C reactive protein (86 mg/L) and erythrocyte sedimentation rate (46 mm/1h) . White blood cells, pus cells and red blood cells were found on routine stool examination. Biochemical panel showed hypoalbuminemia (25.2 g/L) , elevated transaminase (alanine aminotransferase 175 U/L, aspartate transaminase 229 U/L) , hypertriglyceridemia (4.41 mmol/L) , and hyperferritinemia (>1 650.0 μg/L) . Magnetic resonance enterography revealed the intestinal wall thickening and increased enhancement in parts of illeum and colon. Capsule endoscopy revealed multiple ulcers in jejunum. Colonoscopy showed multiple ulcers in colon and the pathological examination revealed chronic inflammation in mucosa of terminal ileum and colon, which was combined with partial necrosis and ulceration. Some phagocytes were seen in bone marrow smears. The patient was given multiple diagnoses, including hemophagocytic lymphohistiocytosis, Crohn's disease, sepsis, epilepsy, severe malnutrition, and hypoproteinemia. The pediatric Crohn's disease activity index (PCDAI) was 37.5. Genetic testing identified a hemizygotic mutation of c.910G>T chrX:123022501 p.G304X in XIAP. The parents had no such mutation. The patient showed response to infliximab with oral intake of mercaptopurine and corticosteroids, and had remission with PCDAI of 0. There were 9 relevant articles (Chinese 0 English 9), which showed 33.3% XIAP deficient patients manifested with inflammatory bowel disease(IBD), who might have other manifestations such as hemophagocytic lymphohistiocytosis or splenomegaly simultaneously or sequentially. Those patients showed poor response to monotherapy.@*Conclusion@#XIAP deficient patients have various clinical manifestations. Genetic testing is important to those male pediatric IBD patients who have the complicated symptoms or little response to standard therapy.

High expression of XIAP and Bcl-2 may inhibit programmed cell death in glioblastomas / Alta expressão de XIAP e Bcl-2 pode inibir morte celular programada em glioblastomas

ABSTRACT Glioblastoma (GBM) is the most malignant glioma and represents 29% of all brain tumors. Tumorigenesis is intimately connected with characteristics acquired in the physiologic pathway of cellular death. Objective: In the present study, the expression of anti-apoptotic (XIAP and Bcl-2) and apoptotic (cytochrome C, caspase 9, APAF-1), caspase 3 and the Smac/DIABLO genes related to the apoptosis pathway were evaluated in 30 samples of glioblastoma. Methods: The gene expression was evaluated in 30 glioblastomas (WHO grade IV) and compared to 10 white matter control samples with real-time PCR. Results and Conclusion: There were higher expressions of XIAP (p = 0.0032) and Bcl-2 (p = 0.0351) in the glioblastoma samples compared to the control samples of normal brain. These results raise the question of whether Bcl-2 and XIAP genes can be responsible for the inhibition of programmed cell death in glioblastomas. Moreover, they provide additional information capable of allowing the development of new target therapy strategies.

RESUMO O glioblastoma (GBM) é o glioma mais maligno e representa 29% de todos os tumores cerebrais. A tumorigênese está intimamente ligada à características adquiridas na via fisiológica de morte celular. Objetivo: Avaliar a expressão de genes anti-apoptóticos (XIAP e Bcl-2) e apoptóticos (citocromo C, a caspase 9, APAF-1), caspase 3 e SMAC/DIABLO, relacionados à apoptose, em 30 amostras de tecido de pacientes com glioblastoma. Métodos: A expressão gênica foi avaliada em trinta glioblastomas e comparada a dez amostras controles de substância branca por PCR em tempo real. Resultados e Conclusão: Houve maior nível de expressão de XIAP (p = 0,0032) e Bcl-2 (p = 0,0351) em comparação com as amostras controle, de cérebro normal. Estes resultados levantam a questão de que os genes Bcl-2 e XIAP podem ser responsáveis pela inibição da morte celular programada em glioblastomas, além disso, proporcionam informação adicional capaz de permitir o desenvolvimento de novas estratégias de terapia alvo.

Treatment of paclitaxel combined with oxaliplatinchemotherapy to the non-small cell lung cancer mouse model / 基础医学与临床

Objective To investigate the therapeutic effect of paclitaxel plus oxaliplatin chemotherapy to the transplanted non-small cell lung cancer of nude mice and the effect to the apoptosis protein expression of PDCD5 and XIAP with mice model.Methods A tumor-bearing mice were randomly divided into blank group, normal saline group, oxaliplatin group, paclitaxel group, paclitaxel plus oxaliplatin group.The gene expression of PDCD5 and XIAP was assayed by real-time quantitative PCR(q-PCR).The apoptosis related PDCD5 and XIAP protein were detected by Western blot.Finally, the tumor weight of each group was measured for statistical analysis.ResultsThe mRNA expression of PDCD5 was highest and the gene expression of XIAP was lowest in paclitaxel plus oxaliplatin group(P<0.01).The expression of PDCD5 protein was highest and the expression of XIAP protein was lowest in paclitaxel plus oxaliplatin group (P<0.01).Finally, compare the tumor weight of each group, paclitaxel plus oxaliplatin group has the least mass(P<0.01).Conclusions Paclitaxel plus oxaliplatin group chemotherapy significantly increases PDCD5 expression and reduce XIAP expression.Meanwhile, paclitaxel plus oxaliplatin chemotherapy can significantly reduce the tumor weight of happened non-small cell lung cancer.

Differential Expression of Transcriptional Variants of XAF1 in Colorectal Neoplasms / 胃肠病学

Background:The tumor suppressor,X-linked inhibitor of apoptosis(XIAP)-associated factor 1(XAF1)is a XIAP-binding protein that antagonizes the anti-caspase activity of XIAP,thereby enhancing apoptosis. Transcriptional variants of XAF1 have been detected in various tumor cells,however,the expression profile of these transcriptional variants in colorectal neoplasms remains unclear. Aims:To investigate the expressions of XAF1 and its transcriptional variants in different colorectal tissues and their roles in tumorigenesis and development of colorectal neoplasms. Methods:Samples of colorectal cancer and paired adjacent tissue,hyperplastic polyp,adenomatous polyp,and normal colorectal mucosa were collected from surgical operation or endoscopic biopsies. XAF1 protein expression was detected by immunohistochemistry and Western blotting,and the transcriptional variants of XAF1 were detected by RT-PCR. Results:Compared with normal colorectal mucosa,the expression level of XAF1 protein in nucleus was significantly reduced( P 0. 05)in hyperplastic polyp,adenomatous polyp,and cancerous tissue,and the overall expression level of XAF1 protein was decreased(P <0. 05). XAF1A protein expression in cancerous tissue was significantly reduced when compared with the paired adjacent tissue(P < 0. 05). mRNA expressions of three transcriptional variants of XAF1,XAF1A,XAF1B and XAF1C were all significantly lower in neoplastic tissues than in normal mucosa(P < 0. 05). Conclusions:XAF1 and its transcriptional variants are differentially expressed in colorectal neoplasms and normal colorectal mucosa. These changes occurred initially in adenomatous polyp accompanied by a redistribution of XAF1 from nucleus to cytoplasm. Post-transcriptional modification may affect XAF1 gene function.

Effects of baicalin on mitochondria apoptotic pathway of trophoblast cells in a preeclampsia rat model / 中华围产医学杂志

Objective To explore the effects of baicalin in the treatment of a preeclampsia (PE) rat model by detecting the expression of X-linked inhibitor of apoptosis protein (XIAP) and cysteine containing aspartate-9 (Caspase-9) and observing the ultrastructure of mitochondria in trophoblast cells.Methods Forty-eight pregnant Wistar rats were randomly divided into two groups:12 in the control group and 36 in the PE model group.The PE model was established with subcutaneous injection of l-nitro arginine methyl ester with 100 mg/kg per day from the 13th day of pregnancy.Beginning from the 16th day of pregnancy,the PE rats were injected with different doses of baicalin till cesarean section,and divided into three groups:non-intervention PE model group treated with saline (NIP group),low-dose baicalin intervention group (IDB group) at 50 mg/kg per day,and high-dose baicalin intervention group (HDB group) at 100 mg/kg per day.The rat tail artery blood pressure and 24-h urine protein level were measured at pregnant day 10,16 and 20.The levels of XIAP and Caspase-9 in placenta were measured by immunohistochemistry.The ultrastructure of mitochondria of trophoblast cells of the rat placenta was observed under electron microscope.T test,F test and LSD-t test were applied for statistical analysis.Results (1) On pregnant day 10,no significant differences were observed in rat tail artery systolic blood pressure,diastolic blood pressure and 24-h urine protein level between the control group and PE model group (all P＞0.05).On pregnant day 16 and 20,the systolic blood pressure,diastolic blood pressure and 24-h urine protein level of NIP,IDB and HDB groups were significantly higher than those of control group [pregnant day 16:systolic blood pressure:(137.74±5.21),(136.15±4.86),(138.28±4.79) and (110.57±3.79) mmHg (1 mmHg=0.133 kPa),diastolic blood pressure:(89.58 ± 5.50),(88.45 ± 8.59),(89.42 ± 6.29) and (80.28 ± 7.36) mmHg,24-h urine protein:(7.78 ± 0.45),(7.53 ± 0.54),(7.86± 0.57) and (6.45 ± 0.56) mg;pregnant day 20:systolic blood pressure:(145.26 ± 4.67),(131.28 ± 4.34),(130.93 ± 5.33) and (110.40 ± 6.92) mmHg,diastolic blood pressure:(89.87±6.55),(85.34±7.33),(84.64±7.36) and (80.19±7.34) mmHg,24-h urine protein:(11.18±0.42),(9.65±0.54),(9.06±0.56) and (6.31 ±0.45) mg] (all P＜0.01).On pregnant day 20,the systolic and diastolic blood pressure and 24-h urine protein level in IDB and HDB groups were significantly lower than in NIP group (all P＜0.05),but showed no significant differences between IDB and HDB groups (allP＞0.05).(2) Compared with NIP group,the expression of XIAP in control group,IDB and HDB groups were significantly increased(210.39±0.78,180.56±0.82,195.36±0.96 and 192.84± 1.06,all P＜0.01).There was no significant difference in the expression of XIAP between IDB and HDB groups (P=0.66).The expression of Caspase-9 in control group,IDB and HDB groups were significantly decreased compared with NIP group (210.36±0.55,195.53±0.96,198.42± 1.01 and 185.25±0.64,all P＜0.01).There was no significant difference in the expression of Caspase-9 between IDB and HDB groups (P=0.65).Ultrastructure of mitochondria in NIP group showed different degrees of damage,matrix swelling,and mitochondrial cristae bresk or disappearance.In IDB group,mitochondrial matrix swelling was not obvious,and mitochondrial cristae were visible.In HDB group,mitochondrial cristae were neat and clear.Conclusions Baicalin may play an important role in the treatment of preeclampsia by reversing the trophoblast apoptosis and improving the ultrastructure of mitochondria through its regulation of XIAP expression and downregulation of Caspase-9 expression.

Effect of Matrine on the expression of X-linked inhibitor of apoptosis protein in human rhabdomyosarcoma RD cells / 中华实用儿科临床杂志

Objective To investigate the effect of Matrine on the expression of X-linked inhibitor of apoptosis protein (XIAP) in human rhabdomyosarcoma RD cells in vitro.Methods Cultured human rbabdomyosarcoma RD cells were divided into Matrine intervention groups (0.5 g/L,1.0 g/L and 1.5 g/L) and a control group.The proliferation-inhibition rates in RD cells treated with different concentrations of matrine were detected by methylthiazolyl blue colorimetric assay.Flow cytometry analysis was performed for the apoptosis rates of RD cells.Reverse transcription-polymerase chain reaction analysis was used to measure the XIAP mRNA expression.Results There was a significant difference in the proliferation-inhibition rates [0.5 g/L Matrine group:(15.84 ± 2.58)％,1.0 g/L Matrine group:(23.13 ±4.19)％,1.5 g/L Matrine group:(30.32 ±3.02)％,and the control group:(8.92 ± 1.23)％];apoptotic rates [0.5 g/L Matrine group:(12.33 ± 1.15)％,1.0 g/L Matrine group:(16.67 ± 0.99)％,1.5 g/L Matrine group:(25.33 ± 1.91)％,and the control group:(9.47 ± 0.96)％];XIAP mRNA expression(0.5 g/L Matrine group:0.633 ± 0.046,1.0 g/L Matrine group:0.441 ± 0.055,1.5 g/L Matrine group:0.326 ± 0.065,control group:0.794 ±0.029)in RD cells among 0.5 g/L,1.0 g/L,1.5 g/L Matrine groups and the control group (F =14.15,83.37,50.57,all P ＜ 0.05).The proliferation-inhibition and apoptotic rates in RD cells were gradually increased with the increasing Matrine concentration.The expression of XIAP mRNA was significantly decreased in different Matrine groups compared with the control group,exhibiting a dose-dependent manner.Conclusions Matrine can inhibit the proliferation of RD cells and induce the apoptosis in a dose-dependent manner,which may be related to the down-regulated XIAP mRNA.

Effects of TSG on H2O2-induced Apoptosis and Expressions of XIAP and p53 in HUVECs / 华中科技大学学报(医学版)

Objective To study the effects of tetrahydroxy stilbene glucoside(TSG)on H2O2‐induced apoptosis of human umbilical vein endothelial cells (HUVECs)and on the expressions of X‐linked inhibitor of apoptosis protein (XIAP)and p53.Methods HUVECs were cultured in vitro and divided into 6 groups:control group ,300 μmol/L H2O2 group ,1 μmol/L TSG+ 300 μmol/L H2O2 group ,10 μmol/L TSG+300 μmol/L H2O2 group ,100 μmol/L TSG+ 300 μmol/L H2O2 group ,30μmol/L Embelin+ 10 μmol/L TSG+ 300 μmol/L H2O2 group.The morphology of apoptotic cells was observed by Hoechst 33258 staining.The mRNA and protein expressions of XIAP ,p53 ,Caspase‐3 were detected by RT‐PCR and Western blotting , respectively.Results The number of apoptotic cells and the expression level of p53 were significantly increased while the ex‐pression level of XIAP was dramatically decreased in H2O2 group as compared with control group.The expression level of p53 and the number of apoptotic cells were down‐regulated while the expression level of XIAP was up‐regulated after treatment with 10 or 100 μmol/L TSG when compared with H2O2group.Moreover ,compared with those in 10 μmol/L TSGgroup ,the number of apoptotic cells and the expression of Caspase‐3 were significantly enhanced after pretreatment with 30 μmol/L Embelin for 6 h.Conclusion TSG can inhibit H2O2‐induced apoptosis of HUVECs by down‐regulating the expression level of p53 and up‐reg‐ulating the expression level of XIAP.

Construction of X IAP-3′UTR-luciferase reporter vector and its activity analysis / 国际检验医学杂志

Objective To construct the recombinant X-linked inhibitor of apoptosis protein(XIAP) gene 3′untranslational region (3′UTR)-luciferase reporter vector ,and analyze the microRNA(miRNA) which possibly regulate the expression of XIAP gene . Methods Polymerase chain reaction (PCR) was employed to amplify X IA P-3′UTR sequences from human cDNA ,in which luciferase reporter vector pGL3-Ctrl was inserted ,and the recombinant vector pGL3-Ctrl/XIAP was gained .Target Scan 6 .2 soft-ware was adopted to predict the miRNA which possibly combined with the X IA P-3′UTR .pGL3-Ctrl/XIAP recombinant plasmids and the miRNA were co-transfected into A549 cells ,and the X IA P-3′UTR-luciferase activity was measured .Results Confirmed by digestion and DNA sequencing ,the X IA P-3′UTR-luciferase reporter recombinant was successfully constructed .Prediction of miRNA target sites indicated that X IA P gene may be the target of miR-200b ,miR-200c and miR-429 .Compared with miRNA mim-ic ctrl group ,miR-200b ,miR-200c and miR-429 significantly reduced the luciferase activity of pGL 3-Ctrl/XIAP with statistically significant difference(P<0 .05) .Conclusion X IA P-3′UTR-luciferase reporter vector is successfully constructed .miR-200b ,miR-200c and miR-429 can obviously decrease the luciferase activity .

Sensitizing effect of chemotherapy of isorhapontigenin through down-regulating expression of X-linked inhibitor of apoptosis / 中国药学杂志

OBJECTIVE: To observe the inhibitor effects of isorhapontigenin (ISO) on the proliferation of bladder cancer cells and expression of X-linked inhibitor of apoptosis (XIAP), and the sensitizing effect of chemotherapy of docetaxel to bladder cancer cells In ISO. METHODS: The ATPase assay and anchorage-independent growth assay were used to detect the cytotoxicity and proliferative capacity of ISO on the T24T bladder cancer cell line, respectively. The reverse transcription polymerase chain reaction (RT-PCR) and Western-blotting methods were used to detect XIAP gene expression after pretreated with ISO. Pretreatment with different concentration of docetaxel and ISO on T24T bladder cancer cell, ATPase methods were performed to measure in vitro cell viability. And the morphological changes and apoptosis rates were detected by phase contrast microscope and flow cytometry assay, respectively. RESULTS: ISO could exhibit significant inhibitory effects on human bladder cancer cell growth through ATPase assay and anchorage-independent growth assay. Further proved by Western-blotting and RT-PCR methods, the protein and mRNA level of XIAP gene in T24T cells were both significantly decreased. After pretreatment with ISO, the IC50 of T24T cells for docetaxel was (1.02 ± 0.38) nmol · L-1, significantly lower than that docetaxel without ISO, which was (8.78 ± 1.32) nmol · L-1 for 24 h( P < 0.01). Compared with the control group, the typical apoptosis morphological changes were observed in the T24T cells pretreatment with ISO combined with docetaxel. And the index of apoptosis was (21.07 ± 2.79)% at 2.5 nmol · L-1 docetaxel, markedly lower than those of T24T cells pretreatment with 2.5 nmol · L-1 docetaxel and ISO, which was (49.59 ± 5.67)% (P < 0.01). CONCLUSION: ISO could exhibit significant inhibitor) effects on human bladder cancer cell proliferation. And the XIAP gene could be down-regulated significantly by ISO, which could increase docetaxel-induced apoptosis and cytotoxic activity significantly.

Expression of XIAP and Smac in human non-small-cell lung carcinoma(NSCLC)and the relationship with clinical significance and prognosis / 中国肿瘤临床

Objective:To investigate the expression of XIAP and Smac in human non-small-cell lung carcinoma (NSCLC) and the relationship with clinical significance and prognosis. Methods:Immunohistochemical staining was performed to determine the ex-pression of X-linked inhibitor of apoptosis protein (XIAP) and second mitochondria-derived activator of caspase (Smac) in 70 cases of NSCLC and 70 cases of non-cancerous adjacent lung tissues. Results:XIAP is mostly present (59/70) in tumor tissues with 16 high ex-pressions, whereas only five high expressions in non-cancerous adjacent lung tissues are observed (52/70). The statistical difference of these two sets of data is significant (Z=-5.484, P0.05). The Kaplan-Meier analysis results show that survival by XIAP and Smac protein in NSCLC has no significant effect (P>0.05). Conclusion:XIAP and Smac are expressed in NSCLC and noncancerous adjacent lung tissues, and the differences in their expression levels is significant. The deterioration of NSCLC results in apoptosis/anti-apoptotic synchronized with tumor cell proliferation. The expression levels of XIAP and Smac in NSCLC are not related with the prognosis.

Molecular mechanism of berberine induced apoptosis in chemoresistant acute lymphocytic leukemia cell line EU-4 / 白血病·淋巴瘤

Objective To study the molecular mechanism of berberine induced apoptosis in chemoresistant EU-4 acute lymphocytic leukemia cells.Methods EU-4 cells were treated with 0,10 and 100 μmol/L berberine for 72 h.The apoptosis induced by berberine was detected by flow cytometry.The expression of Caspase-3,PARP and X-linked inhibitor of apoptosis protein (XIAP) were determined by Western bolt assay.Caspase-3 activity was measured using microplate reader.After transfected with XIAP siRNA,the apoptosis was detected by flow cytometry.Results After treated with 0,10 and 100 μmol/L berberine for 72 h,the apoptosis rates of EU-4 cells were (9.08±1.20) ％,(22.36±2.16) ％ and (59.81±4.17) ％,respectively.Berberine induced potent apoptosis in a dose-dependent manner.The apoptosis involved activation of Caspase-3.The Caspase-3 activities were 1.70±0.25,1.92±0.10 and 2.89±0.25,respectively.Berberine inhibited XIAP expression in a dose-and time-dependent manner (P ＜ 0.05).Down-regulation of XIAP by siRNA increased apoptosis of EU-4 cells.The apoptosis rates were (9.23±1.66) ％ and (22.15±0.63) ％.Conclusion Berberine could induce apoptosis of EU-4 cells,and inhibition of XIAP leading to Caspase-3 activation is responsible for the apoptotic effect on EU-4 cells.

Identification and Modification of XIAP Gene in Rat Adipose-Derived Stem Cells / 天津医药

Objective To investigate the feasibility of genetically modified X-linked inhibitor of apoptosis protein (XIAP) of rat adipose-derived mesenchymal stem cells (ADSCs) by isolating and cultivating rat ADSCs in vitro. Methods ADSCs were isolated from rat groin fat pads by collagenaseⅠdigestion under sterile condition. ADSCs were passaged and amplified with 10%FBS DMEM. The multi-differentiation potential of ADSCs was verified by cultivated with differentiation medium. XIAP expression plasmid was transfected into ADSCs. The anti-apoptotic ability of XIAP transduction was detect-ed by Western blotting assay. Results ADSCs were mainly spindle-shaped and whirlpool-shaped arranged. Results of flow cytometry showed that there were higher expressions of CD29, CD44, CD90 and CD105 in ADSCs, which differentiated into lipocytes, chondrocytes and osteoblasts under specific conditions. There is XIAP gene modified adipose-derived mesenchy-mal stem cells Band in the corresponding molecular mass of PVDF membrane area. Conclusion ADSCs were isolated from rat subcutaneous fat pads and were easily cultivated, passaged and amplified. ADSCs can differentiate into osteoblasts, chon-drocytes and adipocytes under specific conditions, which are better resource for being used in cell therapy and tissue engi-neering.

XIAP modified adipose-derived stem cells improve cardiac function following myocardial infarction / 中华胸心血管外科杂志

Objective We want to study the therapeutic efficiency of autologous ADMSC transplantation in myocardial infarction.And we try to find out a good way to improve the therapeutic efficiency by using the combination of gene therapy and cell therapy.Anti-apoptotic protein XIAP was selected to fight against the ischemic environment of myocardic infaction.Methods ADMSC was isolated from rat inguinal fat tissue.ADMSC was cultivated with DMEM.XIAP experession plasmid was elertco-transduced to ADMSC.The anti-apototic function of XIAP was tested by serum stavation induced apotosis.The method of ligation of the left anterior descending artery was used to prepare the Myocardial infarction model.Then rats were randomly separated into three groups to receive direct epicardial injections of normal saline,or ADMSCs cell suspension or XIAP modified ADMSCs cell suspension at five sites in central zones of myocardial infarction and border zone.Cardiac function and the infarct size were evaluated 4 weeks after ADMSCs transplantation.Results West blotting suggest that,XIAP over-expression block serum starvation induced apotosis.It showed that there are significant statistic difference among XIAP modified ADMSC transplantation group,ADMSC transplantation group and control group 4 weeks after myocardial infarction (P ＜ 0.05).Left ventricular ejection fraction(LVEF) showed a significant improvement in ADMSCs transplantation group compared to control group (P ＜0.05).Left ventricular end systolic diameter(LVDs) and left ventricular end diastolic diameter(LVDd) of ADMSCs group were smaller than control group(P ＜ 0.05).The area of myocardial infarction was significantly reduced in the ADMSCs transplantation group compared to the saline group(P ＜0.05).Compared to ADMSCs transplantation group,effect of the XIAP modified ADMSC in rats with myocardial infarction is more obvious.The reduction of LVEF of XIAP modified ADMSCs group was signific antly lower(9％) than the ADMSCs group(16％) (P ＜ 0.01).Infarction area in XIAP modified ADMSCs group(3.26 ±0.95)％ was smaller than ADMSCs group(5.17 ±2.03)％ (P ＜0.05).Conclusion Autologus ADMSC transplantation is an efficient therapeutic tool in myocardial infarction therapy.Over expression of XIAP can partly inhibit lowserum induced apotosis of ADMSC in vitro,and it can improve left ventricular function better in vivo.Over expression XIAP of ADMSC can improve the therapeutic efficiency compare to ADMSC transplantation.

Construction of recombinant plasmid siRNA of XIAP and its biological role / 中国医师杂志

Objective To construct siRNA expression vector of XIAP,and study its effect on XIAP expression in Hep3B cells. MethodsThree XIAP siRNA sequences were designed,synthesized,and cloned to pRNAT-U6.1/Neo.The successfully constructed recombinant plasmid was determined by sequence analysis,and will be transfected into Hep3B.The best interference plasmid were analyzed by RTPCR,Western blot,and immunohistochemistry.Results The plasmid of pRNAT-U6.1/Neo-XIAP was constructed successfully,the trans-fected with different plasmid of siRNA XIAP can lower significantly XIAP.Conclusions The siRNA vector of XIAP gene was constructed successfully.It will be a basis for the study of XIAP function in apoptosis regulation of the Hepatoma cells.

Inhibitors of Apoptosis Proteins Expression and Their Prognostic Significance in Colorectal Carcinoma

BACKGROUND: The expression of the inhibitor of apoptosis proteins (IAPs) family has not been fully investigated in colorectal carcinomas. This study investigated IAP expression in colorectal carcinomas and assessed their prognostic significance. METHODS: Livin, XIAP, and SMAC/DIABLO expression was assessed by immunohistochemistry in 159 colorectal carcinomas. Correlations between protein expression and clinicopathological features were evaluated. The survival data analysis was estimated according to the Kaplan-Meier method. RESULTS: Increased expression of IAPs in cancer tissues compared to surrounding nonneoplastic counterparts was observed in 67 cases (42.1%) for Livin, 50 cases (31.4%) for XIAP, and 68 cases (42.8%) for SMAC. A significant correlation was found between Livin expression and tumor differentiation, and SMAC expression and tumor location. The recurrence-free and overall survival of patients with low Livin expression were inferior to those of patients with high Livin expression (p=0.054 and 0.095, respectively). High XIAP expression was significantly associated with shorter progression-free survival (p= 0.041). CONCLUSIONS: Our study demonstrated that altered expression of IAP family members, including Livin, XIAP, and SMAC, is frequent in colorectal carcinoma. This result suggests that high Livin expression and low XIAP expression may be a favorable prognostic implication related to patient survival.

Expression of X-linked Inhibitor of Apoptosis Protein in Neoplastic Thyroid Disorder

X-linked inhibitor of apoptosis protein (XIAP) is associated with tumor genesis, growth, progression and metastasis, and acts by blocking caspase-mediated apoptosis. In the present study, we sought to evaluate the expression patterns of XIAP in various neoplastic thyroid disorders and determine the association between XIAP expression and clinicopathologic factors. Expression of XIAP was evaluated with immunohistochemical staining using monoclonal anti-XIAP in 164 specimens of conventional papillary thyroid carcinoma (PTC) and 53 specimens of other malignant or benign thyroid tumors. XIAP positivity was observed in 128 (78%) of the 164 conventional PTC specimens. Positive rates of XIAP expression in follicular variant PTC, follicular, medullary, poorly differentiated, and anaplastic thyroid carcinoma specimens were 20%, 25%, 38%, 67%, and 38%, respectively. Six nodular hyperplasia specimens were negative and 1 of 7 follicular adenomas (8%) was positive for XIAP. Lateral neck lymph node metastases were more frequent in patients negative for XIAP expression (P = 0.01). Immunohistochemical staining for XIAP as a novel molecular marker may thus be helpful in the differential diagnosis of thyroid cancer. Moreover, high XIAP expression in conventional PTC is strongly associated with reduced risk of lateral neck lymph node metastasis.