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.

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.

Relationship between iASPP expression with clinicopathological features and prognosis in patients with nasopharyngeal carcinoma / 重庆医学

Objective To investigate the expression change of inhibitory member of the ASPP family(iASPP) and its relationship with clinicopathological features and prognosis in the patients with nasopharyngeal carcinoma(NPC).Methods One hundred and thirty-three cases of nasopharyngeal carcinoma confirmed by pathology in the hospital were selected,at the same time 30 cases of nasopharyngeal chronic mucositis biopsy paraffin blocks previously collected in the pathology department served as the control group.The immunohistochemical staining was used to compare the expression of iASPP protein in the two groups and the relationship between the expression of iASPP with the clinicopathologic characteristics and prognosis in the patients with NPC was analyzed.Results The positive expression of iASPP protein in the nasopharyngeal carcinoma tissue was 72.18％,which was higher than 20.00％ in the control group(P＜0.05);the positive expression of iASPP protein in the nasopharyngeal carcinoma tissue had a certain relationship with the primary tumor range and lymph node metastasis(P＜0.05);92 cases of nasopharyngeal carcinoma were followed up,including 63 cases of iASPP protein positive expression and 29 cases of iASPP protein negative expression,the 5-year survival rate in the patients with iASPP protein positive expression was 71.43％,which in the patients with iASPP protein negative expression was 86.21％,the difference was not statistically significant(P＞0.05).Conclusion The iASPP expression in the nasopharyngeal carcinoma tissue is up-regulated,which is related to the lymph node metastasis and the primary tumor range,but its influence on the patient's prognosis is not significant.

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.

Analysis on the expression of Smac and Survivin in precancerous lesions of gastric ulcer / 国际检验医学杂志

Objective To investigate the expression of pro-apoptotic factor (Smac) and Survivin in gastric ulcer tissue.Methods Selected the 80 cases of gastric ulcer patients as the research object in the first people′s hospital of neijiang,in which no precancerous lesions of 40 cases of gastric ulcer (N group),40 cases of precancerous lesions of gastric ulcer(Y group),two groups of patients were Smac mRNA and Survivin mRNA were detected by using PCR method,immunohistochemical SP method of Smac and Survivin in specimens of table detect.Gastric ulcer patients were treated by triple therapy,and the apoptosis index was detected by TUNEL.Results Smac in N group(++) and (+++) in the expression accounted for 82.5％ was higher than that in Y group accounted for 47.5％,Survivin in group N (++) and (+++) in the expression accounted for 15.0％ was significantly lower than Y group accounted for 35.0％;And Smac mRNA in the N group relative expression the amount was significantly higher than that of Y group,while the expression of Survivin mRNA in the N group were significantly lower than Y group;N group the apoptosis index of the triple therapy after treatment than before treatment significantly decreased,the difference was statistically significant(P<0.05).Conclusion The clinical application of Smac and Survivin can be used as an auxiliary diagnostic index for the diagnosis of precancerous lesions in patients with gastric ulcer.Patients with gastric ulcer without precancerous lesion treated by triple therapy which can effectively control the apoptosis index of patients,improve the survival rate of patients.

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.

The effect of c-IAP1 on radiosensitivty of lung cancer cells / 国际生物医学工程杂志

Objective To investigate the correlation between the level of apoptosis protein 1 (c-IAP1) and the radiosensitivity of lung cancer cells.Method The survival rate and proliferation of the lung cancer cells lines (A549,H460,H1299,H358,HCC827,H1650) from six human were detected by thiazolyl blue tetrazolium bromide (MTT) and cell colony formation assay.The DNA damage effects of radiation on lung cancer cells were detected by comet assay.The expressions of c-IAP1 protein and its mRNA were determined by Western blot and real-time quantitative PCR.Results The results of MTT and colony formation showed that the radiosensitivity of different lung cancer cells was also different,among which H358 and H460 cells had the highest radiosensitivity than that of H1650 and HCC827 cells,and H1299 and A549 cells had the weakest radiosensitivity.The results of comet assay showed that six kinds of lung cancer cells were suffered by DNA damage after radiation,and the DNA damage of H358 cells was most serious.The results of Western blot and real-time quantitative PCR showed that the c-IAP1 protein level was negatively correlated with the radiosensitivity of lung cancer cells.The higher the c-IAP1 protein level,the weaker the radiosensitivity of cells.The radiosensitivity was also affected by Smac protein levels.Conclusions c-IAP1 may be a selective target gene in mediating the radiosensitivity of lung cancer cells and this paper may contribute to the study of radioresistance and radiosensitization of cancer cell.

The effect of c-IAP1 on radiosensitivty of lung cancer cells / 国际生物医学工程杂志

Objective To investigate the correlation between the level of apoptosis protein 1 (c-IAP1) and the radiosensitivity of lung cancer cells.Method The survival rate and proliferation of the lung cancer cells lines (A549,H460,H1299,H358,HCC827,H1650) from six human were detected by thiazolyl blue tetrazolium bromide (MTT) and cell colony formation assay.The DNA damage effects of radiation on lung cancer cells were detected by comet assay.The expressions of c-IAP1 protein and its mRNA were determined by Western blot and real-time quantitative PCR.Results The results of MTT and colony formation showed that the radiosensitivity of different lung cancer cells was also different,among which H358 and H460 cells had the highest radiosensitivity than that of H1650 and HCC827 cells,and H1299 and A549 cells had the weakest radiosensitivity.The results of comet assay showed that six kinds of lung cancer cells were suffered by DNA damage after radiation,and the DNA damage of H358 cells was most serious.The results of Western blot and real-time quantitative PCR showed that the c-IAP1 protein level was negatively correlated with the radiosensitivity of lung cancer cells.The higher the c-IAP1 protein level,the weaker the radiosensitivity of cells.The radiosensitivity was also affected by Smac protein levels.Conclusions c-IAP1 may be a selective target gene in mediating the radiosensitivity of lung cancer cells and this paper may contribute to the study of radioresistance and radiosensitization of cancer cell.

IAP-1 gene silencing enhanced the radiosensitivity of liver cancer SMMC-7721 cells / 中华肝胆外科杂志

Objective To explore the effect of cellular inhibitor of apoptosis protein1 (cIAP-1) gene on the radiosensitivity of SMMC-7721 cells.Methods We silenced cIAP-1 expressions by the shRNA technology,and then we detected the changes of cell proliferation,cell cycle and cell apoptosis by CCK8 as-say,Western blot,qRT-PCR and flow cytometry after the radiotherapy.Results The cell proliferation rate of liver cancer SMMC-7721 cells at different radiation doses of 1 Gy,4 Gy,7 Gy and 10 Gy was detected.Comparing with control group (pGCsi-H1-control),the cell proliferation in cIAP-1 silencing group (pGCsiH1-shRNA) was significantly reduced at various radiation doses,and the effect was dose-dependent (P ＜ 0.05).G1/G0 phase arrest was observed after radiation (P ＜0.01),and the proportion of cells in S phase was significantly reduced compared with control (P ＜ 0.01).Compared with control group,G1/G0 phase arrest was detected (P ＜ 0.05),and the percentage of cell apoptosis was increased significantly in cIAP-1 silencing group (P ＜ 0.05).Conclusion cIAP-1 silencing can enhance the radiosensitivity of liver cancer cells,and inhibit cell proliferation by promoting the anti-tumor effect of radiation.

Helicobacter pylori Eradication Downregulates Cellular Inhibitor of Apoptosis Protein 2 in Gastric Carcinogenesis

BACKGROUND/AIMS: To evaluate the expression of cellular inhibitor of apoptosis protein 2 (cIAP2) during gastric carcinogenesis after Helicobacter pylori (HP) infection and after HP eradication. METHODS: We divided non-cancer patients into four groups according to the status of HP infection and atrophic gastritis (AG)/intestinal metaplasia (IM). We compared cIAP2 mRNA expression among these four groups and patients with HP-positive early gastric cancer (EGC) by using real-time polymerase chain reaction (PCR). We evaluated the expression of cIAP2 messenger RNA (mRNA)/protein by using real-time PCR/immunohistochemistry and the degree of apoptosis with a terminal deoxynucleotidyl transferase-mediated nick end labeling assay before and 12 months after endoscopic submucosal dissection (ESD) in HP-positive EGC patients, regardless of whether they had undergone eradication therapy. RESULTS: The expression of cIAP2 mRNA was significantly higher in the groups with HP(+), AG/IM(+), and HP-positive EGC than in the control, HP(+), and AG/IM(−) groups (p<0.005). In the HP eradication group, the expression of cIAP2 mRNA/protein significantly decreased (p=0.006) and apoptosis increased at the 12-month follow-up after ESD. In the HP noneradication group, the aforementioned changes were not found during the same follow-up period. CONCLUSIONS: The expression of cIAP2 increased during gastric carcinogenesis after HP infection; HP eradication in the patients who had undergone ESD for EGC reversed overexpression of cIAP2 and suppressed cell apoptosis.

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 cIAP1 down-regulation mediated by shRNA on biological behavior of human ovarian cancer cells / 实用肿瘤学杂志

[Abstratc] Objective The function of cIAP1 in the progression of ovarian cancer has not been clarified . This study is to explore the involvement of cIAP 1 in regulating biological behaviors of ovarian cancer cells by u-sing RNA interference(RNAi)technology.Mte hods The short hairpin RNA plasmid targeting cIAP1 was con-structed and transfected into Skov 3 cells.The levels of cIAP1 mRNA and protein were investigated by RT -PCR and Western Blot respectively .MTT assay and flow cytometry were used to evaluate cell proliferation and apopto-sis.R esults The rate of cIAP1 transfection was 74.7%performed by flow cytometric analysis .cIAP1 expression was significantly down -regulated at both mRNA and protein levels ,which resulted in a decrease of cell prolifera-tion and invasion capability in vitro .Conclusion This study implies that cIAP 1 might play an important role in the progression of ovarian cancer ,and it could be a potential target for therapeutic anti -cancer drugs .

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.

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.

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.

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 .

Effect of siRNA targeting Livin gene on biological characteristics of human prostate cancer PC3 cells / 中华泌尿外科杂志

Objective To observe the effect of RNAi targeting Livin gene on biology characteristics such as apoptosis and proliferation in human prostate cancer cells.Methods siRNA expression vector targeting Livin gene was constructed and transfected into human prostate cancer cell line PC3.The expressions of Livin mRNA and protein were detected by real-time PCR and Western-blot,cell apoptosis and cell cycle were assayed by flow cytometry,proliferation and colony formation were detected by MTT and colony formation assay,and the tumor growth in vivo was observed in nude mice.Results After transfection,downregulation of Livin mRNA and protein expression in PC3 cells was observed (P＜0.01).Compared with the control group,the proliferation of cancer cells was inhibited significantly (P＜0.01) and the apoptotic ratio was (26.5±3.3) ％ (P＜0.01).The Caspase3 activity increased obviously (P＜0.05),and the experimental group showed a decreased colony formation rate (P＜0.01).The tumor volume of xenografts in nude mouse in experimental and control group was (1.79± 0.07) and (4.40 ± 0.06) cm3 respectively (P ＜ 0.01).Conclusions The siRNA recombinant expression vector targeting Livin gene was constructed and can knockdown the expression of Livin mRNA and protein.It can inhibit PC3 cell proliferation,induce apoptosis and inhibit tumor growth in vivo.

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.

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.