Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; metabolism ; Protein Isoforms ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo study the effect of adenovirus vector-mediated siRNA targeting vascular endothelial growth factor(VEGF) on apoptosis and the expression of survivin in K562 cells.

METHODSK562 cells were infected with recombinant adenovirus Ad5-VEGFsi for 72 hours as experimental group (K562/Ad5-VEGFsi), and empty vector group (K562/Ad5) and blank control group (K562) as controls. VEGF mRNA and survivin mRNA expression were determined by RT-PCR. The protein levels of VEGF and survivin were measured by ELISA and Western blot, respectively. The apoptosis of K562 cells was detected by flow cytometry.

RESULTSThe levels of VEGF and survivin mRNA expression in experimental group cells were significantly decreased (P < 0.01). The protein concentration of VEGF in experimental group supernatant was (1121 +/- 15) pg/ml, being lower than that in empty vector group \[(1290 +/- 28) pg/ml\] and black control group \[(1303 +/- 28) pg/ml\] (P < 0.01), and the level of survivin protein in experimental group (0.26 +/- 0.11) was significantly reduced compared with that in blank control group (0.74 +/- 0.10) (P < 0.01). The apoptosis rate of K562/Ad5-VEGFsi cells (16.45 +/- 0.14)% was higher than those of K562/Ad5 cells (3.54 +/- 0.17)% and K562 cells (2.56 +/- 0.20)% (P < 0.01).

CONCLUSIONSVEGF can up-regulate the expression of survivin. After inhibition of VEGF by RNAi, the expression of survivin is decreased subsequently and the rates of cell apoptosis are increased.

Apoptosis ; drug effects ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; K562 Cells ; RNA, Small Interfering ; genetics ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo observe the effect of survivin antisense oligonucleotides (ASODN) on survivin and Smac gene expression in ovarian cancer SKOV3 cells and explore the role of survivin and Smac genes in ASODN-induced ovarian cancer cell cycle alteration and apoptosis and its molecular mechanisms.

METHODSSurvivin ASODN was introduced via Lipofectamine(TM)2000 into SKOV3 cells, whose growth activity was detected subsequently with MTT assay. The apoptosis index, cell cycle and changes in survivn and Smac protein expression were determined using flow cytometry. The changes in survivn and Smac mRNA expression were detected by RT-PCR.

RESULTSIn comparison with the control cells, cells transfected with difference concentrations of survivin ASODN exhibited significantly inhibited growth, and 48 h after the transfection, the IC(50) was about 600 nmol/L. After a 48-hour transfection of the cells with 600 nmol/L ASODN, the apoptosis index significantly decreased (t=6.3671, P<0.05), and the cell percentage in (1)/G(0) phase increased (t=10.3832, P<0.01), resulting also in significantly down-regulated survivin mRNA and protein expressions (t=3.5031, P<0.05; t=7.8146, P<0.01) and up-regulated Smac mRNA and protein expressions (t= 2.8011, P<0.05; t= 11.3917, P<0.01).

CONCLUSIONASODN against survivin can induce apoptosis of ovarian cancer cell line SKOV3 and results cell growth arrest in G(1)/G(0) phase by up-regulating Smac and down-regulating survivin expression. Survivin and Smac are closely correlated in their action on SKOV3 cell cycle and apoptosis, which is one of the important mechanisms of ovarian cancer development.

Apoptosis ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Inhibitor of Apoptosis Proteins ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Microtubule-Associated Proteins ; genetics ; metabolism ; Mitochondrial Proteins ; genetics ; metabolism ; Oligonucleotides, Antisense ; genetics ; metabolism ; Ovarian Neoplasms ; genetics ; metabolism ; physiopathology

This study was purposed to investigate the mechanism of C-reactive protein (CRP) on proliferation of U266 cells. The human multiple myeloma cell line U266 was incubated with human CRP (0, 5, 10, 20 mg/L) for 24 hours, then the proliferation level of U266 cells was detected by using blood analyser. The mRNA expressions of survivin and HSP90alpha were examined by RT-PCR. The results showed that the proliferation ratio was increased, as compared with the control group (p<0.05); furthermore, the mRNA levels of survivin and HSP90alpha were up-regulated in proportion to the increased CRP concentrations. There was significant correlation between expression of survivin and HSP90alpha (r=0.737, p<0.0001) in incubated cells. It is concluded that CRP can stimulate the proliferation of MM cells directly by up-regulating the expression of survivin and HSP90alpha in MM cells. CRP can be regarded as a potential target for MM treatment.
Apoptosis ; C-Reactive Protein ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; HSP90 Heat-Shock Proteins ; metabolism ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; RNA, Messenger ; genetics

OBJECTIVEThe precise mechanism of glucocorticoid-induced apoptosis has not yet been elucidated. Survivin, a member of the inhibitors of apoptosis protein family, correlates with inhibition of apoptosis, proliferation, angiogenesis and multiple drugs resistance. This study aimed to investigate the variation of the survivin gene expression in apoptosis induced by dexamethasone (Dex) in the human T-lineage acute lymphoblastic leukemia (ALL) cell line, CEM-WT cells.

METHODSThe logarithmically growing CEM cells cultured in vitro (cell density 2 x10(5)/mL) were exposed to 0.1, 0.5, 1, 5, and 10 microM Dex, then were collected 24, 48 and 72 hrs later. Untreated CEM cells were used as Controls. The cell viability was determined by trypan blue dye exclusion. Apoptosis was evaluated by morphology and flow cytometry. Survivin protein and gene were analyzed by Western Blot and RT-PCR.

RESULTSCEM cells growth was obviously inhibited by 0.1, 0.5, 1, 5, and 10 microM Dex from 48 hrs. The inhibition effect was dose- and time-dependent. CEM cells treated with Dex (> or = 5 microM) exhibited typical apoptotic features. The apoptosis increased after 5 microM Dex treatment in a time-dependent manner, with the apoptosis percentage increasing from 14.9% (12 hrs) to 46.2% (48 hrs). Compared with that of the Control group, the expression of survivin protein was down-regulated, with the expression rate of 54.6%, 45.5%, 15.8% and 9.7% respectively at 12, 24, 48 and 72 hrs after 5 microM Dex treatment. 5 microM Dex treatment also resulted in a decrease of survivin mRNA expression. The survivin mRNA expression was 76.4%, 67.3%, 55.0%, 49.9%, 38.3% and 18.3% of the Control respectively at 6, 12, 24, 48 and 72 hrs after Dex treatment.

CONCLUSIONSApoptosis induced by Dex in CEM cells is associated with downregulation of the survivin expression.

Apoptosis ; drug effects ; Cell Line, Tumor ; Dexamethasone ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; Leukemia-Lymphoma, Adult T-Cell ; metabolism ; pathology ; Microtubule-Associated Proteins ; analysis ; genetics ; Neoplasm Proteins ; analysis ; genetics

OBJECTIVETo construct a recombinant adenovirus vector bearing dual-survivin short hairpin RNA (shRNA).

METHODSDual-survivin shRNA was designed and synthesized respectively, both inserted into adenovirus DNA. The recombinant adenovirus vector was confirmed via both sequencing and restriction digestion analysis, and then linearized and transfected into the HEK 293 cell line to generate recombinant adenoviruses.

RESULTSThe recombinant adenovirus vector was constructed and the target sequence was obtained.

CONCLUSIONThe construction of the recombinant adenovirus vector provides a basis for the research of potential gene therapy for prostate cancer.

Adenoviridae ; genetics ; Cell Line ; DNA Restriction Enzymes ; metabolism ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Polymerase Chain Reaction ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

OBJECTIVETo construct a recombinant adenovirus of survivin vector and provid valuable reference for gene therapy of laryngeal cancer.

METHODSThe survivin gene was cloned by PCR. After confirmation by enzyme restriction analysis and sequencing, the gene and the adenovirus vector were recombined together to construct the recombinant adenovirus vector. The recombinant adenovirus vector was confirmed via both sequencing and digestion restriction analysis, and then linearized and transfected into the HEK 293 cell line to generate recombinant adenovirus.

RESULTSThe sequence analysis demonstrated that the survivin gene sequence was the same as published in the literature, suggesting that a recombinant adenovirus vector has been successfully constructed.

CONCLUSIONSA survivin recombinant adenovirus has been successfully constructed.

Adenoviridae ; genetics ; Genetic Vectors ; HEK293 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Plasmids ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

To explore the expression of livin gene in acute non-lymphocytic leukemia (ANLL) cells and its clinical significance, the mRNA level of livin gene in 46 ANLL adult patients were measured by using reverse transcription polymerase chain reaction (RT-PCR). Other 10 healthy adults were selected as normal controls (NC), HL-60 cell line was employed as positive control. The results showed that the mRNA level of livin gene in ANLL patients was significantly higher than that in NC, while it decreased in patients with complete remission (CR). In relapsed patients, the level of livin mRNA increased again. In ANLL patients, the CR rate of patients with livin positive was lower than that of patients with livin negative (p<0.05). It is concluded that overexpression of livin gene may play a synergic role in the pathogenesis of ANLL and associates with CR rate in ANLL. It seems that high expression of livin gene may be used as a marker of poor prognosis in acute non-lymphocytic leukemia.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Adolescent ; Adult ; Aged ; Biomarkers, Tumor ; genetics ; metabolism ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; Prognosis ; RNA, Messenger ; genetics ; metabolism ; Young Adult

This study was aimed to investigate the effect of bortezomib alone or combined with arsenic trioxide on the apoptosis of Jurkat cells and expression of livin mRNA. The Jurkat cells were cultured and treated with different concentrations of bortezomib, arsenic trioxide or their combination for 24 hours. Then, the expression of livin mRNA was detected by RT-PCR, the cell proliferation was analyzed with MTT assay and flow cytometry. The results showed that 5 - 25 nmol/L bortezomib could effectively inhibit Jurkat cells in a dose-dependent manner, the group of bortezomib combined with arsenic trioxide showed more inhibitory effect on Jurkat cells than the effect of bortezomib alone or arsenic trioxide alone on Jurkat cells. The expression of livin mRNA in Jurkat cells decreased in a dose-dependent manner after treated with bortezomib, which was downregulated significantly after combined treatment. It is concluded that bortezomib and arsenic trioxide can induce apoptosis by inhibiting the expression of livin mRNA in Jurkat cells. The combination of bortezomib with arsenic trioxide displays a synergistic effect.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Boronic Acids ; administration & dosage ; pharmacology ; Bortezomib ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Jurkat Cells ; Neoplasm Proteins ; genetics ; metabolism ; Oxides ; pharmacology ; Pyrazines ; administration & dosage ; pharmacology ; RNA, Messenger ; genetics

OBJECTIVETo determine the expression of osteopontin (OPN) and survivin in prostate cancer tissue, and study their correlation and roles in tumor invasion and metastasis.

METHODSThe expressions of OPN and survivin in prostate cancer tissue, prostate hyperplasia tissue and normal prostate tissue were determined by RT-PCR and immunohistochemistry.

RESULTSThe positive expression rates of OPN mRNA and protein in prostate cancer tissue [76.1% (35/46) and 69.6% (32/46)] were significantly correlated to survivin expression [67.4% (31/46) and 67.4% (31/46)] (P<0.05). The expressions of OPN and survivin were related to the tumor grade and clinical stages (P<0.05). OPN and survivin were not found in prostate hyperplasia and normal prostate tissues.

CONCLUSIONOPN and survivin may play important roles in the progression of prostate cancer and can be potential markers for invasion and metastasis of prostate cancer. OPN and survivin might play synergetic roles in prostate carcinogenesis.

Adult ; Aged ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Osteopontin ; biosynthesis ; genetics ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism