Apoptosis ; Hep G2 Cells ; Humans ; RNA, Small Interfering ; X-Linked Inhibitor of Apoptosis Protein ; genetics

OBJECTIVETo investigate the prevalence of mutations and sequence variations in X-linked inhibitor of apoptosis (XIAP) gene among Chinese pediatric patients with hemophagocytic lymphohistiocytosis (HLH).

METHODSSixty-five children who were diagnosed with HLH between January 2009 and December 2012 (case group), as well as 70 healthy children (control group), were enrolled in the study. The exons of XIAP gene (1-1, 1-2, 2-6) were amplified by PCR and directly sequenced. The genotypic and allelic frequencies of single nucleotide polymorphism (SNP) were analyzed.

RESULTSNone of the HLH patients showed mutations in these exons of XIAP gene. Only one nonsynonymous SNP, rs5956583 located in exon 5, was observed, but there were no significant differences in the genotypic and allelic frequencies of this SNP between the case and control groups (P>0.05).

CONCLUSIONSHLH caused by XIAP mutations may be rare in children. SNP rs5956583 of XIAP gene may have little contribution to the development of childhood HLH.

Child ; Child, Preschool ; Female ; Humans ; Lymphohistiocytosis, Hemophagocytic ; genetics ; Male ; Mutation ; Polymorphism, Single Nucleotide ; X-Linked Inhibitor of Apoptosis Protein ; genetics

OBJECTIVETo explore the effect of silence of Pin1 expression on hyperoxia-induced apoptosis in alveolar epithelial cells A549.

METHODSA549 cells were divided into four groups: control, hyperoxia, negative lentivirus and Pin1-shRNA hyperoxia. The hyperoxia group was exposed to a mixture of 95%O2 and 5%CO2 for 10 minutes. Then cells were cultured in a closed environment. After 24 hours, the changes of morphology were observed under an inverted microscope. Cell apoptosis was detected by flow cytometry (FCM). The expression of X-linked inhibitor of apoptosis protein (XIAP) and Caspase-9 were detected by immunohistochemistry. The production of reactive oxygen species (ROS) and cellular mitochondria membrane potential (△Ψm) were determined by fluorescence microscopy.

RESULTSUnder the inverted microscope, the A549 cells grew slowly and the changes in morphology of the cells were most obvious in the hyperoxia and negative lentivirus groups. The changes in morphology of A549 cells were obviously improved in the Pin1-shRNA hyperoxia group. The FCM results showed that the apoptosis rate of A549 cells increased, Caspase-9 expression increased, XIAP expression decreased, mitochondrial ROS production increased and mitochondrial membrane potential decreased in the hyperoxia and negative lentivirus groups compared with the control group (P<0.05). Compared with the hyperoxia and negative lentivirus groups, the apoptosis rate of A549 cells decreased, Caspase-9 expression decreased, XIAP expression increased, mitochondrial ROS production decreased and mitochondrial membrane potential increased in the Pin1-shRNA hyperoxia group (P<0.05), although the levels of the indexes did not reach to those of the control group.

CONCLUSIONSSilencing of Pin1 could suppress hyperoxia-induced apoptosis of A549 cells.

Apoptosis ; Caspase 9 ; genetics ; Humans ; Hyperoxia ; pathology ; Membrane Potential, Mitochondrial ; NIMA-Interacting Peptidylprolyl Isomerase ; Peptidylprolyl Isomerase ; physiology ; Reactive Oxygen Species ; metabolism ; X-Linked Inhibitor of Apoptosis Protein ; genetics

To study the expression of X-linked inhibitor of apoptosis protein (XIAP) and cell apoptosis in vitro model of acute pancreatitis (AP), we carried out experiments to stimulate AR42J cell line with caerulein (10(-8) mol/L) for 12 hours, then collected cells at various time points (0 h, 4 h, 8 h, 12 h, and 24 h, respectively). We then observed the morphologic changes of AR42J cells with the stimulation of caerulein with electronic microscope. The gene expression of XIAP, caspase-3 and caspase-9 was detected using real-time fluorescence quantitative polymerase chain reaction (FQ-PCR), and the protein expression of XIAP was assessed by western blot. The activation of nuclear factor-kappa B (NF-kappaB) was measured by flow cytometry (FCM). With the stimulation of caerulein, the expression of XIAP and the NF-kappaB activation could first decrease and then increase, but the change of caspase-3 and caspase-9 expressions were opposite. XIAP may inhibit the cell apoptosis in rat pancreatic acinus AR42J cell lines at first with the stimulation of caerulein, then NF-kappaB can upgrade the expression of XIAP and increase the cell apoptosis.
Acinar Cells ; cytology ; metabolism ; Animals ; Apoptosis ; physiology ; Cell Line ; Ceruletide ; pharmacology ; NF-kappa B ; metabolism ; Pancreas ; cytology ; metabolism ; Pancreatitis ; metabolism ; Rats ; X-Linked Inhibitor of Apoptosis Protein ; genetics ; metabolism

OBJECTIVE: To investigate the down-regulation effect of antisense oligonucleotides (AS ODNs) targeting X-chromosome-linked inhibitors of apoptosis (XIAP) on hep-2 cells apoptosis and radiotherapy sensitivity in vitro. METHOD: G4 AS ODN was transfected into cultured hep-2 cells which received radiation 6 hours later. Twenty-four hours after radiation, cells were observed under inverted phase contrast microscope and fluorescence microscope. Apoptosis rate, cell viability, expression of XIAP mRNA and protein were tested. RESULT: In cultured hep-2 cells, G4 AS ODN down-regulated XIAP mRNA expression. Furthermore, the protein expression of XIAP and the cell viability decreased too. In contrast to that, the scrambled control ODN caused minor XIAP loss and less cell inhibition. In addition, G4 AS ODNs activated Hep-2 cells after the radiation of 4 Gy Co60 ray. CONCLUSION XIAP is a viable target for cancer therapy in human laryngeal neoplasms. In cultured Hep-2 cells, AS ODN targeting XIAP can down-regulate protein expression of XIAP, induce cell apoptosis and enhance the radiotherapy sensitivity.
Apoptosis ; drug effects ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Oligonucleotides, Antisense ; pharmacology ; X-Linked Inhibitor of Apoptosis Protein ; genetics ; pharmacology

This study was purpose to investigate the expression of XIAP mRNA in chronic myeloid leukemia (CML) and to explore its significance in the advance and prognosis of CML. The chromosomal karyotype analysis and detection of XIAP mRNA were performed by the technique of chromosomal R banding and real time PCR in 71 patients with CML and 10 healthy controls. The results showed that there was a significant increase of XIAP mRNA expression in accelerated and blastic phase of the CML, compared with the patients in chronic phase (t = 9.10, 9.30, p < 0.01). Moreover, the difference of XIAP mRNA expression level was not statistically significant in different karyotype groups. It is concluded that the XIAP gene expression in accelerated and blastic phases of CML patients obviously increases, XIAP gene is closely related to the advance of CML.
Adult ; Case-Control Studies ; Female ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; pathology ; Male ; Prognosis ; X-Linked Inhibitor of Apoptosis Protein ; genetics

Amino Acid Sequence ; Base Sequence ; Child ; Flow Cytometry ; methods ; Genes, X-Linked ; Hematopoietic Stem Cell Transplantation ; Humans ; Lymphocytes ; metabolism ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; genetics ; therapy ; Lymphoproliferative Disorders ; diagnosis ; genetics ; therapy ; Mutation ; Transplantation, Homologous ; X-Linked Inhibitor of Apoptosis Protein ; deficiency ; genetics ; metabolism

OBJECTIVETo study the expression of the inhibitor of apoptosis (IAP) gene XIAP in prostate cancer and the relationship between its expression and its clinical stage or Gleason grade.

METHODSXIAP mRNA expression was detected by RT-PCR in three prostate cancer cell lines (PC-3, DU-145, LNCaP), neoplastic prostate tissues and normal prostate tissues. Immunohistochemical SP method was used to examine the expression of XIAP protein in 56 neoplastic prostate tissue specimens.

RESULTSXIAP gene was not expressed in normal prostate tissues, but highly expressed in prostate cancer cell lines PC-3, DU-145, LNCaP. Thirty of the 56 (53.6%) tumor samples were positive XIAP protein, and only 12 (21.5%) paratumor samples were positive XIAP protein (P < 0.01), XIAP positivity not correlated with tumor stage or Gleason grade (P > 0.05).

CONCLUSIONXIAP may be involved in the development of prostate cancer and play an important role in human prostate carcinogenesis. It is likely to be used as a target of prostate cancer therapy.

Aged ; Aged, 80 and over ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Male ; Neoplasm Staging ; Prostatic Neoplasms ; metabolism ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; X-Linked Inhibitor of Apoptosis Protein ; biosynthesis ; genetics

Adenocarcinoma ; metabolism ; pathology ; Apoptosis ; drug effects ; Baculoviral IAP Repeat-Containing 3 Protein ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromones ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Morpholines ; pharmacology ; Ovarian Neoplasms ; metabolism ; pathology ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; Ubiquitin-Protein Ligases ; X-Linked Inhibitor of Apoptosis Protein ; genetics ; metabolism

This study was purposed to explore the mechanisms of drug resistance of HL-60/ADR cells and to compare the reversal drug-resistance effects of antisense oligonucleotides (AS ODN) of XIAP (X-linked inhibitor of apoptosis protein) and AS ODNs of MRP (multidrug resistance-associated protein) by use alone or in combination. Reverse transcription-PCR and Western blot were applied to detect the expression of XIAP, BCL-2, MRP and MDR1 in mRNA and protein levels of HL-60 cells and HL-60/ADR cells, respectively. Fully phosphorothioated AS ODN of XIAP and MRP was delivered into HL-60/ADR cells with Lipofectamine 2000 in the form of liposome-ODN complexes alone or in combination. CCK-8 cell viability assay was used to determine the effect of AS ODN of XIAP and MRP used alone or in combination on the chemotherapy sensitivity of HL-60/ADR cells to daunorubicin (DNR). Reverse transcription-PCR and Western blot were applied to examine the changes of XIAP, MRP in mRNA and protein levels respectively. The results showed that MRP and XIAP were both significantly higher in HL-60/ADR cells than those in HL-60 cells. AS ODN of XIAP and MRP down-regulated the expression of XIAP and MRP in HL-60/ADR cells and increased the sensitivity of HL-60/ADR cells to DNR, respectively. AS ODN of XIAP + MRP did not enhance the inhibition expression of XIAP in HL-60/ADR cells but increased the sensitivity of HL-60/ADR cells to DNR significantly as compared with AS ODN of XIAP (P < 0.05). AS ODN of XIAP + MRP did not increase the concentration of DNR nor enhanced the inhibition expression of MRP in HL-60/ADR cells but increased the sensitivity of HL-60/ADR cells to DNR significantly (P < 0.05), as compared with AS ODN of MRP. It is concluded that both XIAP and MRP may be involved in the drug resistance mechanisms of HL-60/ADR cells. Drug-resistance of HL-60/ADR cells can be reversed significantly when antisense oligonucleotides of XIAP and MRP were used in combination.
Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; HL-60 Cells ; Humans ; Multidrug Resistance-Associated Proteins ; biosynthesis ; genetics ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; X-Linked Inhibitor of Apoptosis Protein ; biosynthesis ; genetics