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

OBJECTIVETo explore the expression of X-linked inhibitor of apoptosis protein (XIAP) in Tca8113 cell, and to investigate its relationship to the chemoresistance. METHODS; The Tca8113 cell line was cultured by IMDM and the concentration of Pingyangmycin (PYM) added to Tca8113 cell line was increased gradually and continually, which was to induce the PYM-resistance in Tca8113 cell line. The sensitivity of Tca8113 cell to PYM and expression of XIAP were measured with methyl thiazolyl tetrazolium (MTT) chromatometry and reverse transcription-polymerase chain raction (RT-PCR). The XIAP level in the cells and its chemoresistance to PYM were analyzed by linear regression.

RESULTSThe IC50 of Tca8113-1-10 group and Tca8113-10-10 group were(12.758 +/- 0.030), (18.986 +/- 0.150) microg x mL(-1) respectively. The IC50 of Tca8113-1-20 group and Tca8113-10-20 group increased to (26.302 +/- 0.072), (35.294 +/- 0.115) microg x mL(-1) respectively. There was a relation between XIAP and the drug-resistance in Tca8113 cell.

CONCLUSIONXIAP may play an important role in the chemoresistance which might serve as a new therapeutic target for oral squamous cell carcinoma.

Apoptosis ; Bleomycin ; analogs & derivatives ; Carcinoma, Squamous Cell ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Humans ; X-Linked Inhibitor of Apoptosis Protein

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.
Apoptosis ; Colorectal Neoplasms ; Disease-Free Survival ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Intracellular Signaling Peptides and Proteins ; Mitochondrial Proteins ; Prognosis ; Proteins ; Statistics as Topic ; X-Linked Inhibitor of Apoptosis Protein

BACKGROUND: Cisplatin (cis-diaminedichloroplatinum, CDDP) is a widely used chemotherapeutic agent for the treatment of many cancers. However, initial resistance to CDDP is a serious problem in treating these cancers. Vitis coignetiae Pulliat (Meoru in Korea) have shown anti-nuclear factor kappa B and anti-epidermal growth factor receptor activities in cancer cells. METHODS: In this study, in order to seeking an approach to increase the anti-cancer effects of CDDP with natural products. Here, we investigated anthocyanins isolated from Vitis coignetiae Pulliat (anthocyanidins isolated from meoru, AIMs) can enhance anti-cancer effects of cisplatin (CDDP) in stomach cancer cells. The cell viability of SNU-1 and SNU-16 cells after treated with AIMs and CDDP were analyzed by MTT assay. The expressions of Akt and X-linked inhibitor of apoptosis protein (XIAP) proteins were examined by western blot in AIMs- and CDDP-treated cells. RESULTS: We found that AIMs enhanced anticancer effects of CDDP, which activity was additive but not synergistic. AIMs suppressed Akt activity of the cancer cells activated by CDDP. AIMs also suppressed in XIAP an anti-apoptotic protein. CONCLUSIONS: This study suggests that the anthocyanins isolated from fruits of Vitis coignetiae Pulliat enhanced anti-cancer effects of CDDP by inhibiting Akt activity activated by CDDP.
Anthocyanins* ; Biological Products ; Blotting, Western ; Cell Survival ; Cisplatin* ; Fruit* ; Humans* ; Stomach Neoplasms* ; Vitis* ; X-Linked Inhibitor of Apoptosis Protein

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

BACKGROUND: alpha-Lipoic acid (alpha-LA) has been studied as an anticancer agent as well as a therapeutic agent for diabetes and obesity. We performed this study to evaluate the anticancer effects and mechanisms of alpha-LA in a lung cancer cell line, A549. MATERIALS AND METHODS: alpha-LA-induced apoptosis of A549 cells was detected by fluorescence-activated cell sorting analysis and a DNA fragmentation assay. Expression of apoptosis-related genes was analyzed by western blot and reverse transcription-polymerase chain reaction analyses. RESULTS: alpha-LA induced apoptosis and DNA fragmentation in A549 cells in a dose- and time-dependent manner. alpha-LA increased caspase activity and the degradation of poly (ADP-ribose) polymerase. It induced expression of endoplasmic reticulum (ER) stress-related genes, such as glucose-regulated protein 78, C/EBP-homologous protein, and the short form of X-box binding protein-1, and decreased expression of the anti-apoptotic protein, X-linked inhibitor of apoptosis protein. Reactive oxygen species (ROS) production was induced by alpha-LA, and the antioxidant N-acetyl-L-cysteine decreased the alpha-LA-induced increase in expression of apoptosis and ER stress-related proteins. CONCLUSION: alpha-LA induced ER stress-mediated apoptosis in A549 cells via ROS. alpha-LA may therefore be clinically useful for treating lung cancer.
Acetylcysteine ; Apoptosis ; Blotting, Western ; Cell Death ; Cell Line ; DNA Fragmentation ; Endoplasmic Reticulum ; Flow Cytometry ; Lung Neoplasms ; Obesity ; Reactive Oxygen Species ; Thioctic Acid ; X-Linked Inhibitor of Apoptosis Protein

In Korea, chestnut production is increasing each year, but consumption is far below production. We investigated the effect of chestnut extracts on antioxidant activity and anticancer effects. Ethanol extracts of raw chestnut (RCE) or chestnut powder (CPE) had dose-dependent superoxide scavenging activity. Viable numbers of MDA-MD-231 human breast cancer cells, DU145 human prostate cancer cells, and AGS human gastric cancer cells decreased by 18, 31, and 69%, respectively, following treatment with 200 microg/mL CPE for 24 hr. CPE at various concentrations (0-200 microg/mL) markedly decreased AGS cell viability and increased apoptotic cell death dose and time dependently. CPE increased the levels of cleaved caspase-8, -7, -3, and poly (ADP-ribose) polymerase in a dose-dependent manner but not cleaved caspase-9. CPE exerted no effects on Bcl-2 and Bax levels. The level of X-linked inhibitor of apoptosis protein decreased within a narrow range following CPE treatment. The levels of Trail, DR4, and Fas-L increased dose-dependently in CPE-treated AGS cells. These results show that CPE decreases growth and induces apoptosis in AGS gastric cancer cells and that activation of the death receptor pathway contributes to CPE-induced apoptosis in AGS cells. In conclusion, CPE had more of an effect on gastric cancer cells than breast or prostate cancer cells, suggesting that chestnuts would have a positive effect against gastric cancer.
Apoptosis ; Breast ; Breast Neoplasms ; Caspase 8 ; Caspase 9 ; Cell Death ; Cell Survival ; Ethanol ; Humans ; Korea ; Prostatic Neoplasms ; Stomach Neoplasms ; Superoxides ; X-Linked Inhibitor of Apoptosis Protein

OBJECTIVETo investigate the effect of proteasome inhibitor bortezomib on proliferation, apoptosis of K562 cells and the expression of XIAP.

METHODSK562 cells were treated with bortezomib at different concentration. Cell proliferation was analyzed by WST-1 assay, cell apoptosis by flow cytometry and TUNEL, XIAP mRNA expression from 5 - 100 nmol/L by RT-PCR, and XIAP protein expression by SP immunohistochemistry.

RESULTSK562 cells were treated with bortezomib at different concentrations for 24 h respectively, the cells growth was significantly inhibited with inhibition rates from (13. 6 ± 0. 2)% to (81. 4 ± 0. 1)%, respectively, being markedly higher than that of control (1. 2 ± 0. 1)% (P < 0.05). IC(50) was 24. 6 nmol/L of bortezomib treated for 24 h. When K562 cells were treated with 30 nmol/L of bortezomib for 12 - 48 h, the inhibition rates were (29. 1 ± 0. 9)% to (59. 8 ± 1. 2)%, respectively, the differences being statistically significant (P < 0.05) between 12 h group and 24 h group, while there was no statistical difference between 24 h, 36 h and 48 h groups. K562 cells treated with 30 nmol/L bortezomib for 24 h showed nuclear condensation, nuclear margination, nuclear fragmentation, cytoplasmic vacuoles and a large number of apoptotic body formation. The apoptotic cells rate was 83. 67% in bortezomib treated group, and 2. 33% in untreated group (P < 0.05). The expression of XIAP mRNA was decreased in a dose-dependent manner, and the expression of its protein was down-regulated.

CONCLUSIONBortezomib can inhibit the proliferation of K562 cells, and induce apoptosis by down-regulating the expression of XIAP, providing the laboratory evidence for the targeted therapy in acute leukemia.

Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Proliferation ; drug effects ; Humans ; K562 Cells ; Leukemia ; metabolism ; Pyrazines ; pharmacology ; X-Linked Inhibitor of Apoptosis Protein ; metabolism

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