OBJECTIVE: To observe the effect of sodium valproate (VPA) on proliferation and apoptosis of human laryngeal carcinoma Hep-2 cells and its associated mechanism. METHOD: Methabenzthiazuron (MTT) was used to observe the proliferation of human laryngeal carcinoma Hep-2 cells treated with various concentrations of VPA at different times. Flow cytometry(FCM) and RT-PCR were used to measure the apoptosis rate and the expression of Survivin mRNA in the Hep-2 cells treated with VPA at 3 mmol/L for different times. RESULT: VPA inhibited growth of Hep-2 cells in a dose-dependent and time-dependent manner (P < 0.01). The apoptosis rate increased after the treatment by VPA at 3 mmol/L. There were significant differences between different time groups (P < 0.01). The expression of Survivin mRNA of Hep-2 cells were decreased in a time dependent manner (3 mmol/L) (P < 0.01). CONCLUSION VPA have obvious growth inhibition and induction of apoptosis on human laryngeal cancer Hep-2 cells, its mechanism is related to decrease the expression of Survivin.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Flow Cytometry ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Survivin ; Valproic Acid ; pharmacology

OBJECTIVE: To observe the effect of programmed cell death 5 (PDCD5) protein on the apoptosis of multiple myeloma KM3 cells induced by dexamethasone and to understand its mechanism. METHODS: The human recombinant PDCD5 (rhPDCD5) protein was added (alone of different concentrations or associated with dexamethasone) into KM3 cells. Cultured together for certain time, the cells were collected for the following experiments: (1)The effect of rhPDCD5 protein and dexamethasone on the apoptotic rate of KM3 cells was determined by flowcytometry (FCM) analysis after the cells were stained by Annexin V-FITC & PI (propidium iodide). (2)Caspase-3 activity of KM3 cells was evaluated by Western blot. (3)The expression of survivin protein in KM3 cells was detected by immunocytochemistry. RESULTS: The apoptotic rate of KM3 cells and the activity of caspase-3 increased significantly, and that treated with rhPDCD5 protein and dexamethasone was higher than that treated with rhPDCD5 protein only. The expression of survivin protein in the rhPDCD5 with dexemethas group was down-regulated, and with the concentration of rhPDCD5 and dexamethasone increasing, the changes was more obviously. CONCLUSION PDCD5 protein can induce the apoptosis of KM3 cells, and accelerate the apoptosis of KM3 cells induced by dexamethasone. PDCD5 protein may reduce the expression of survivin protein and increase activation of caspase-3 to play its role in promoting apoptosis.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; pharmacology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Dexamethasone ; pharmacology ; Drug Synergism ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Multiple Myeloma ; pathology ; Neoplasm Proteins ; pharmacology ; Recombinant Proteins ; pharmacology ; Survivin

OBJECTIVE: To investigate the effects of mTOR inhibitors everolimus (EVE) and gemcitabine (GEM) on the proliferation, apoptosis and cell cycle of diffuse large B-cell lymphoma (DLBCL) cell line U2932, and further explore the molecular mechanisms, so as to provide new ideas and experimental basis for the clinical treatment of DLBCL. METHODS: The effect of EVE and GEM on the proliferation of U2932 cells was detected by CCK-8 assay, the IC₅₀ of the two drugs was calculated, and the combination index (CI=) of the two drugs was calculated by CompuSyn software. The effect of EVE and GEM on apoptosis of U2932 cells was detected by flow cytometry with AnnexinV-FITC/PI staining. Flow cytometry with propidium iodide (PI) staining was used to detect the effect of EVE and GEM on the cell cycle of U2932 cells. Western blot assay was used to detect the effects of EVE and GEM on the channel proteins p-mTOR and p-4EBP1, the anti-apoptotic proteins MCL-1 and Survivin, and the cell cycle protein Cyclin D1. RESULTS: Both EVE and GEM could significantly inhitbit the proliferation of U2932 cells in a time- and dose-dependent manner (r=0.465, 0.848; 0.555, 0.796). According to the calculation of CompuSyn software, EVE combined with GEM inhibited the proliferation of U2932 cells at 24, 48 and 72 h with CI=<1, which had a synergistic effect. After treated U2932 cells with 10 nmol/L EVE, 250 nmol/L GEM alone and in combination for 48 h, both EVE and GEM induced apoptosis, and the difference was statistically significant compared with the control group (P<0.05). The apoptosis rate was significantly enhanced after EVE in combination with GEM compared with single-agent (P<0.05). Both EVE and GEM alone and in combination significantly increased the proportion of cells in G₁ phase compared with the control group (P<0.05). The proportion of cells in G₁ phase was significantly increased when the two drugs were combined (P<0.05). The expression of p-mTOR and effector protein p-4EBP1 was significantly downregulated in the EVE combined with GEM group, the expression of anti-apoptotic proteins MCL-1, Survivin and cell cycle protein cyclin D1 was downregulated too (P<0.05). CONCLUSION EVE combined with GEM can synergistically inhibit the proliferation of U2932 cells, and the mechanism may be that they can synergistically induce apoptosis by downregulating the expression of MCL-1 and Survivin proteins and block the cell cycle progression by downregulating the expression of Cyclin D1.
Humans ; Gemcitabine ; Everolimus/pharmacology* ; Survivin/pharmacology* ; Cyclin D1/pharmacology* ; Myeloid Cell Leukemia Sequence 1 Protein ; Cell Line, Tumor ; Cell Proliferation ; TOR Serine-Threonine Kinases ; Apoptosis ; Apoptosis Regulatory Proteins ; Cell Cycle Proteins ; Lymphoma, Large B-Cell, Diffuse

OBJECTIVE: This study aimed to explore the relationship among expression of Survivin and MRP and drug resistance in NPC. METHOD: Expression of Survivin were detected by immunohistochemistry method in 45 cases of NPC and 24 cases of normal mucous membrane of nasopharynx (NMMN). The relationship between expression of Survivin and pathological factors in NPC were analysized. Expression of Survivin and MRP were detected in 31 patients of NPC with paclitaxel resistance and 20 patients of NPC without paclitaxel resistance. The relation- ship among the expression of Survivin or MRP and paclitaxel resistance in NPC were analysized. The paclitaxel resistance cell line, 5-8F-PTX(+); was established by a step-increased method. The expression of Survivin and MRP were detected by western blot in 5-8F-PTX(+) and 5-8F. RESULT: The positive were 71. 1% (32/45) in NPC and 8.33% (2/24) in NMMN. And there were significantly differences between them (P < .05). There were relationship among expression of Survivin and differentiation degree, lymph node metastasis, distant metastasis, and clinic stages of NPC. The positive were 75.9% (31/39) in moderately differentiated NPC and 16.7% (1/6) in lowly differentiated NPC, respectively. There were significantly differences between them (P < 0.05). The positive of Survivin were 83.9% (26/31) in NPC patients with paclitaxel resistance and 45.0% (9/20) in NPC patients without Paclitaxel resistance, respectively. There were significantly differences between them (P < 0.05). The positive of MRP were 87.1% (27/31) in NPC patients with paclitaxel resistance and 40.0% (8/20) in NPC patients without paclitaxel resistance, respectively. There were significantly differences between them (P < 0.05). There were positive correlation between the expression of Survivin and MRP in NPC patients with Paclitaxel resistance. The expression of Survivin and MRP were higher in 5-8F-PTX(+) than in 5-8F. The IC50 of paclitaxel, cDDP, 5-FU and Vincristine were significantly higher in 5-8F-PTX(+) than in 5-8F. CONCLUSION There were relationship among the expression of Survivin and difference, metastasis and TNM stages of NPC. Survivin may serves as a molecular marker for development and progress in NPC. There were relationship among the high expression of Survivin and MRP and increasing of drug resistance in NPC.
Antineoplastic Agents ; pharmacology ; Carcinoma ; Cisplatin ; Drug Resistance, Neoplasm ; Fluorouracil ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Lymphatic Metastasis ; Multidrug Resistance-Associated Proteins ; genetics ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; Nasopharynx ; metabolism ; Paclitaxel ; pharmacology ; Survivin ; Vincristine

OBJECTIVE: To explore the positive expressing rates of oncogene Survivin and tumor-suppressor gene PTEN in transplanted laryngo-carcinoma of nude mice treated by Gold Throat Tablets (GTT) which can improve circulation and remove haemostasis in TCM theory. METHOD: The 32 nude mice seeded with cultured laryngeal carcinoma cells subcutaneously at the back were randomly divided into high, middle, low (according to 6 : 3: 1 proportion of GTT dosing given by intragastric administration) dose groups and blank control groups. The changes on weight and size of tumors originated from these cells were observed for 28 days, and the density of tumors and expression of Survivin and PTEN were examined with tumor sections by immunohistochemical assay after separating tumors from back of nude mice. RESULT: The weight and size of subcutaneous laryngo-carcinoma on backs of high dose group nude mice were both the smallest among all the experimental groups with the average density of tumors as 1.202 g/cm3. The positive expressing rates of Survivin and PTEN both revealed the following tendency that high dose group < middle dose group < low dose group < blank control group. CONCLUSION Six times of regular doses of GTT can prevent overgrowth of laryngo-carcinoma transplanted on nude mice and decrease the excessive expression of oncogene Survivin in the tumor. PTEN, expressing lower than Survivin in all groups, shows a subordinative relationship with it, maybe due to a competition mechanism.
Animals ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Repressor Proteins ; metabolism ; Survivin

OBJECTIVE: To determine the molecular mechanism of reversing multi-drug resistance of K562/AO2 by puerarin. METHODS: Effects of ADR and puerarin on NF-kappaB activity of K562,K562/AO2 were tested by immunofluorescence. The expression of survivin of K562,K562/AO2 was examined by immunocytochemistry. The p-gp expression was detected by flow cytometry. RESULTS: The NF-kappaB activity of K562 was significantly higher than that of K562/AO2. The NF-kappaB activity of K562 treated by ADR was significantly higher than untreated. The NF-kappaB activity of K562 which was pretreated by puerarin and then treated by ADR was much lower than that treated by ADR alone. The NF-kappaB activity of K562/AO2 intervened by puerarin was lower than that unintervened by puerarin.The p-gp and survivin expression of K562/AO2 was significantly higher than K562. The p-gp and survivin expression of K562 treated by ADR was higher than that untreated by ADR. But the p-gp and survivin expression of K562 which was pretreated by puerarin and then treated by ADR was much lower than that not pretreated by puerarin.The p-gp and survivin expression of K562/AO2 intervened by puerarin was lower than that unintervened by puerarin. The expression was negatively correlated to the duration of intervention. The inhibition effect demonstrated time dependence. CONCLUSION The activation of NF-kappaB can increase the expression of p-gp and survivin, which may be part of the molecular mechanism of multi-drug resistance of K562. Puerarin can prevent and stop the multi-drug resistance in K562 and reverse the multi-drug resistance of K562/AO2 to ADR by inhibiting the activity of NF-kappaB and the expression of p-gp and survivin.
ATP Binding Cassette Transporter, Subfamily B ; biosynthesis ; genetics ; Antineoplastic Agents, Phytogenic ; pharmacology ; Drug Resistance, Multiple ; drug effects ; genetics ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; Isoflavones ; pharmacology ; K562 Cells ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; NF-kappa B ; metabolism ; Survivin

The aim of this study was to investigate the effect of Wnt5a on the vincristine (VCR) resistance in human ovarian carcinoma SKOV3 cells and its possible mechanism. The drug-resistant SKOV3/VCR cells were established by stepwise exposure to VCR, and then the SKOV3/VCR cells were stably transfected with specific shRNA interference plasmid vector targeting for Wnt5a. The mRNA expression level of Wnt5a was measured by RT-PCR. CCK-8 assay was used to detect the cell viability of SKOV3/VCR cells. The apoptosis was analyzed by flow cytometry. The protein expression levels of Wnt5a, MDR1, Survivin, β-catenin, Akt, p-Akt(S473), GSK3β and p-GSK3β(Ser9) were detected by Western blot. The result showed that SKOV3/VCR cells had significantly higher protein expression levels of Wnt5a, MDR1, Survivin and β-catenin, phosphorylation levels of Akt and GSK3β, and mRNA expression level of Wnt5a, compared with SKOV3 cells (P < 0.05). WNT5A gene silencing significantly increased the sensitivity of SKOV3/VCR cells to VCR, the IC of VCR being decreased from 38.412 to 9.283 mg/L (P < 0.05), synergistically enhanced VCR-induced apoptosis of SKOV3/VCR cells (P < 0.05), down-regulated the protein expression levels of MDR1, β-catenin and Survivin (P < 0.05), and inhibited phosphorylation of Akt and GSK3β (P < 0.05). Meanwhile, LY294002 (PI3K inhibitor) decreased the protein expression levels of MDR1, β-catenin and Survivin, as well as the phosphorylation levels of Akt and GSK3β in SKOV3/VCR cells (P < 0.05). These results suggest that WNT5A gene silencing reverses VCR resistance in SKOV3/VCR cells possibly through blocking the PI3K/Akt/GSK3β/β-catenin signaling pathway, and thus down-regulating the protein expression levels of MDR1 and Survivin.
ATP Binding Cassette Transporter, Subfamily B ; metabolism ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Female ; Gene Silencing ; Glycogen Synthase Kinase 3 beta ; metabolism ; Humans ; Ovarian Neoplasms ; pathology ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Survivin ; metabolism ; Vincristine ; pharmacology ; Wnt-5a Protein ; metabolism

OBJECTIVE: To explore the feasibility of antisense oligodeoxynucleotide (ASODN) targeting survivin gene for cisplatin resistant human lung adeno-carcinoma xenograft in nude mice. METHODS: Cisplatin resistant cell lines A549/CDDP were cultured routinely with RPMI1640 medium. A549/CDDP cells were subcutaneously implanted in nude mice to establish cisplatin resistant xenograft animal models. After survivin ASODN mediated by cytofectin was directly injected into xenograft in 5 places. The volumes and weight of tumor mass were detected, respectively, and then tumor growth inhibitory rate and tumor growth index were calculated. Reverse transcription-polymerase chain reaction (RT-PCR) and immunochemohistology assay were performed to detect the expression level of survivin mRNA and protein. RESULTS: In mice treated with single ASODN, the tumor growth inhibitory rate and tumor growth index was 35.4% and 4.23+/-0.4456. The difference of the tumor growth inhibitory rate and tumor growth index between blank control group and ASODN group was significant (P<0.05). While combined ASODN with cisplatin,the anticancer efficacy was far more significant and the tumor growth inhibitory rate was enhanced to 63.7%. The tumor growth index, however, reduced to 1.700+/-0.436, which was obviously significant,compared with the cisplatin group and other controls (P<0.05). The anticancer efficacy was even more obvious than that of ASODN group (P<0.05). Significant down-regulation of survivin mRNA and protein level expression in tumor tissues of ASODN group and ASODN and cisplatin group was detected by RT-PCR and immunochemohistology assay, respectively (P<0.05). CONCLUSION Survivin ASODN mediated by cytofectin can inhibit the cisplatin resistant tumor growth by direct intra-tumoral injection. The anticancer efficacy may be associated with the down regulation of survivin expression. ASODN targeting survivin gene can be a supportive therapy to cisplatin resistant lung cancer, while the clinical effective values need further exploration.
Adenocarcinoma ; pathology ; therapy ; Animals ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Female ; Genetic Therapy ; Humans ; Inhibitor of Apoptosis Proteins ; Lung Neoplasms ; pathology ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microtubule-Associated Proteins ; genetics ; therapeutic use ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; therapeutic use ; Repressor Proteins ; Survivin

OBJECTIVE: To investigate the effect of basic fibroblast growth factor (FGF-2)on survivin and subcellular location of Smac in human small cell lung cancer (SCLC) cell NCI-H446. METHODS: Western blot was used to detect the expression of survivin protein induced by FGF-2. The release of Smac from mitochondria to cytoplasm affected by FGF-2 was observed by Western blot and immunofluorescence. Apoptosis of NCI-H446 cells was detected with flow cytometry and Hoechst 33258 staining. RESULTS: The expression of survivin could be up-regulated in response to FGF-2 treatment in NCI-H446 cells, and the level of survivin expression is related to the concentration and time of FGF-2 treatment. FGF-2 could inhibit the release of Smac from the mitochondria to cytoplasm induced by serum starving. FGF-2 could inhibit the apoptosis induced by serum starving. CONCLUSION FGF-2 up-regulates the expression of survivin protein in NCI-H446 cells, and blocks the release of Smac from mitochondria cytoplasm. Survivin and Smac might play important roles in the apoptosis inhibited by FGF-2.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; Cytoplasm ; metabolism ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; Intracellular Signaling Peptides and Proteins ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Microtubule-Associated Proteins ; biosynthesis ; Mitochondria ; metabolism ; Mitochondrial Proteins ; metabolism ; Small Cell Lung Carcinoma ; metabolism ; pathology ; Survivin ; Tumor Cells, Cultured