Objective To investigate the effects of evodiamine (EVO) on Natural Killer (NK) cell-mediated killing in small cell lung cancer (SCLC) cells via affecting baculoviral inhibitor of apoptosis repeat containing 5 (BIRC5). Methods H446 cells and NK-92 cells were treated with EVO at different concentrations, and cell proliferation was detected using the MTT (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay, while cell invasion was assessed using the Transwell^TM assay. NK-92 cells and H446 cells were co-cultured at different effector-to-target ratios to detect the cytotoxicity of NK cells against H446 cells and the level of degranulation in NK-92 cells. Network pharmacology was employed to analyze the potential targets of EVO in the treatment of SCLC, and further validation was conducted to elucidate the mechanism of EVO's action in SCLC. An xenograft tumor model was used to evaluate the effect of EVO on tumor growth. Results Compared with the control group, EVO treatment dose-dependently inhibited the proliferation and invasion of H446 cells, while enhancing the cytotoxicity of NK-92 cells against H446 cells and the level of NK-92 cell degranulation. Network pharmacological analysis revealed that BIRC5 is a core target of EVO in the treatment of SCLC, and EVO suppressed the expression of BIRC5 protein without affecting BIRC5 mRNA expression. In vivo studies demonstrated that EVO inhibited tumor growth in a dose-dependent manner. Conclusion EVO promotes the degradation of BIRC5, thus enhancing the killing effects of NK cells on SCLC cells.
Humans ; Killer Cells, Natural/metabolism* ; Small Cell Lung Carcinoma/genetics* ; Animals ; Lung Neoplasms/genetics* ; Quinazolines/pharmacology* ; Cell Line, Tumor ; Survivin/genetics* ; Cell Proliferation/drug effects* ; Mice ; Mice, Nude ; Mice, Inbred BALB C ; Xenograft Model Antitumor Assays

Rats ; Animals ; Urinary Bladder Neck Obstruction/pathology* ; Survivin/genetics* ; Urinary Bladder/pathology* ; Fibrosis

OBJECTIVE: To study the effect of knocking down fascin on cervical cancer cell proliferation and tumorigenicity in nude mice. METHODS: Cervical cancer CaSki cells were infected with a lentiviral vector carrying fascin siRNA or with a negative control lentivirus, and fascin mRNA and protein expressions in the cells were detected using qRT-PCR and Western blotting. MTT assay was used to determine the proliferation of CaSki cells with fascin knockdown. CaSki cells transfected with fascin siRNA or the control lentiviral vector and non-transfected CaSki cells were inoculated subcutaneously in nude mice, and the volume and weight of the transplanted tumor were measured; Western blotting was used to detect the expressions of proliferating cell nuclear antigen (PCNA), survivin, cyclin dependent kinase 4 (CDK4) and p21 proteins in the tumor xenograft. RESULTS: Infection with the lentiviral vector carrying fascin siRNA, but not the negative control vector, caused significant reductions in the expression levels of fascin mRNA and protein in CaSki cells ( < 0.05). Fascin knockdown resulted in significantly reduced proliferation of CaSki cells ( < 0.05). The nude mice inoculated with CaSki cells with fascin knockdown showed reduced tumor volume and weight, lowered levels of PCNA, survivin and CDK4, and increased expression of p21 protein in the tumor xenograft compared with the control mice. The negative control lentivirus did not affect the proliferation or tumorigenicity of CaSki cells in nude mice or the expression levels of PCNA, survivin, CDK4 or p21 proteins in the xenografts. CONCLUSIONS Knocking down fascin can inhibit the growth and tumorigenicity of cervical cancer cells in nude mice.
Animals ; Apoptosis ; Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cyclin-Dependent Kinase 4 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Female ; Gene Knockdown Techniques ; Genetic Vectors ; Humans ; Mice ; Mice, Nude ; Microfilament Proteins ; genetics ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; Survivin ; metabolism ; Transfection ; Tumor Burden ; Uterine Cervical Neoplasms ; etiology ; pathology

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 study the combined transfection of the siRNA target for Hif-lα and Survivin gene to human NPC CNE-2 cell and its effects on the proliferation and cycle of this cell. METHOD: Combined transfection of the siRNA target for Hif-lα and Survivin gene to human NPC CNE-2 cell, these plasmids were respectively transfected into the same cells. Cell proliferation was detected with MTT assay. The inhibitory effects on target genes were evaluated with RT-PCR and Western Blot at the levels of mRNA and protein, respectively. RESULT: MTT showed that CNE-2 cell proliferation in multi-gene plasmid group was more significantly inhibited than a single gene. The expression of mRNA and protein of two different genes were both decreased in HS group, and the interference effect of multiple genes was better than the single-gene(P<0.01). CONCLUSION HS group could restrain cell proliferation and interference the mRNA and protein expression in nasopharyngeal carcinoma CNE-2 cell, which was better than the other groups.
Apoptosis ; Carcinoma ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; Inhibitor of Apoptosis Proteins ; genetics ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; genetics ; Plasmids ; RNA, Messenger ; RNA, Small Interfering ; Survivin ; Transfection

OBJECTIVE: To investigate the effect of adenovirus-mediated ING4 with RGD on proliferation, apoptosis and cell cycle of human nasopharyngeal carcinoma cell CNE and explore its probable mechanism. METHOD: CNE cells were infected with Ad. RGD-ING4 and adenovirus vector, ING4 gene expression level was detected by RT-PCR and the target protein expression was tested by Western blot. MTT assay was adopted to evaluate the efect of ING4 on cell growth of CNE, Annexin -V-PE/7-AAD Double staining was used to measure the efect of ING4 on apoptosis, and PI staining was used to measure the efect of ING4 on the cell cycle. Differential expression of P21, Bcl-2 and Bax gene was detected by RT-PCR,and Differential expression of Survivin and Caspase 3 protein was detected by Western blot. RESULT: CNE cells were cultured with Ad. RGD-ING4 for 72 h ,the results showed that ING4 was overexpressed in CNE cells ,the growth of CNE cells was obviously inhibited , apoptosis rate was significantly increased and G2/M phase was arrested apparently. The results of RT-PCR showed that Ad. RGD-ING4 significantly down-regulated the Bcl-2 and up-regulates the Bax and P21 expression in CNE cells, and the difference was statistically significant(P < 0.01). Western blot showed that the expression of Survivin was decreased and Cleaved-Caspase 3 was increased. CONCLUSION Ad. RGD-ING4 can play the role of tumor suppressor synergies on nasopharyngeal carcinoma cell CNE by down-regulating Bcl-2, Survivin expression and up-regulating P21, Bax and Cleaved-Caspase 3 expression.
Adenoviridae ; Apoptosis ; Carcinoma ; Caspase 3 ; metabolism ; Cell Cycle ; Cell Cycle Proteins ; genetics ; therapeutic use ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression ; Genetic Vectors ; Homeodomain Proteins ; genetics ; therapeutic use ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; therapy ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Survivin ; Tumor Suppressor Proteins ; genetics ; therapeutic use

OBJECTIVE: To observe wether pSIREN-survivin/shRNA can induce the apoptosis and inhibit it's growth of nude mice xenograft tumor of nasopharyngeal carcinoma cell. METHOD: Subcutaneous tumors in athymic mice were induced by inoculation of NPC and divided into the blank group A, the negative group B,the experimental group C randomly. PBS,the negative and positive pSIREN-survivin/shRNA were injected into the subcutaneous tumors poly site. The inhibition ratio was measured by the tumor size calculation after inoculation. The expression of survivin in the xenograft tumor was observed by RT-PCR and immunohistochemistry. The cell apoptosis in tumor tissues was detected by TEM. The liver and kidney function was tested by blood routine test. RESULT: The inhibition ratio of group C and group B was (52. 11 +/- 1. 03)%, (2. 15 0. 11)% respectively, the inhibition rate in expression level of survivin mRNA was (77. 5+/-2. 03) %, (1. 39+/-0. 14) % respectively. The dyeing of survivin was more shallow in group C, the intensity is also less than group B. Nuclear chromatin was deepened, split into pieces, flake, and nuclear membrane was surrounded to edge. Cytoplasmic color and density were deepened, and organelles such as mitochondria was disappeared, the microscopic changes of cellular apoptosis at the earlier stage were watched, the difference of the function in liver and kidney was not statistically in statistical. CONCLUSION The expression of survivin in xenograft tumor was significantly inhibited by pshRNA-survivin/shRNA,the apoptosis of tumor cells was accelerated and the growth speed of NPC cells in xenograft tumor was retarded. The high expression of nasopharyngeal carcinoma's gene could significantly be silenced by using technology of RNAi, the growth of tumors could be inhibited also. Its a novel treatment that have a good prospect.
Animals ; Apoptosis ; Carcinoma ; Cell Line, Tumor ; Cell Proliferation ; Female ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; Mice ; Mice, Nude ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; pathology ; RNA, Small Interfering ; genetics ; Survivin ; Xenograft Model Antitumor Assays

OBJECTIVE: To investigate the simultaneous inhibition of X-linked inhibitor of apoptosis protein (XIAP) and survivin expression on epithelial-mesenchymal transition (EMT) and invasiion of pancreatic cancer cells Panc-1, and its mechanism. METHODS: On the established human pancreatic cancer cells Panc-1-XS, the expression of XIAP and survivin was inhibited simultaneously. Cell invasion and migration were detected by Transwell chamber experiments and scratch test, and the expression of epithelial marker E-cadherin, mesenchymal markers Slug, phosphatase and tensin homolog deleted on chromosome ten (PTEN) and P-Akt protein was determined by Western blot. RESULTS: Cell invasion and migration of Panc-1-XS cells decreased significantly, accompanied by significantly upregulated protein expression of E-cadherin, and significantly declined protein expression of the Slug, indicating increased mesenchymal-epithelial conversion (MET); and increased protein expression of PTEN, and declined protein expression of P-Akt. CONCLUSION Simultaneously inhibiting the expression of XIAP and survivin can partially reverse EMT phenotype of pancreatic cancer Panc-1 cells, which then significantly reduces the cell invasion and migration of Panc-1 cell lines. This process may be regulated by PTEN/PI3K/Akt signaling pathway.
Antigens, CD ; Cadherins ; metabolism ; Cell Line, Tumor ; Cell Movement ; Epithelial-Mesenchymal Transition ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Neoplasm Invasiveness ; PTEN Phosphohydrolase ; metabolism ; Pancreatic Neoplasms ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Snail Family Transcription Factors ; Survivin ; Transcription Factors ; metabolism ; X-Linked Inhibitor of Apoptosis Protein ; genetics ; metabolism

OBJECTIVE: To investigate the expression and significance of Survivin mRNA in xenotransplanted nasopharyngeal carcinoma treated by paclitaxel combined with radiotherapy. METHOD: Xenotransplanted nasopharyngeal carcinoma was established by CNE-2 cell line, then grouped and treated with paclitaxel, radiotherapy, paclitaxel combined with radiotherapy respectively. Xenotransplanted tumor volume was measured; tumor specimens were confirmed by routine hemotoxylin-eosin staining; apoptosis index was assayed by flow cytometry and Survivin mRNA was detected by one step RT-PCR. RESULT: Xenotransplanted tumor growth was significantly inhibited by paclitaxel combined with radiotherapy and its inhibition rate was 99.3%. Compared to control group, apoptosis index was apparently increased in the other three groups (P<0.05), especially in the combined therapy group (P<0.05). Survivin mRNA expression was obviously decreased in the combined therapy group (P<0.05); whereas there was no difference in its expression among the groups of paclitaxel, radiotherapy, and control group (P>0.05). CONCLUSION Paclitaxel combined with radiotherapy can induce significant killing effect in xenotransplanted nasopharyngeal carcinoma; paclitaxel can enhance the radiosensitivity of xenotransplanted nasopharyngeal carcinoma and its mechanism may rely on the down-regulation of Survivin expression.
Animals ; Brachytherapy ; Cell Line, Tumor ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Mice ; Mice, Nude ; Nasopharyngeal Neoplasms ; metabolism ; therapy ; Paclitaxel ; therapeutic use ; RNA, Messenger ; genetics ; Repressor Proteins ; genetics ; metabolism ; Survivin ; Xenograft Model Antitumor Assays

OBJECTIVE: (1)To obtain the perfusion parameters of hepatocellular carcinomas(HCCs), peritumour livers and normal livers by multi-slice CT(MSCT)and to investigate their characteristics and clinical significances;(2)To investigate the correlation among perfusion parameters, survivin expression, microvessel density(MVD)and pathologic grade of HCCs. METHODS: A total of 31 patients with HCC (5 well-differentiated HCCs, 17 moderately differentiated HCCs, and 9 poorly differentiated HCCs) and 10 normal liver were studied. All underwent CT plain scan, perfusion scan, and conventional enhancement scan of the whole liver using 16-slice spiral CT (Philips Brilliance 16). Perfusion parameters were obtained by time-density curves (TDC) of region of interest (ROI) through the perfusion scans. Tissue sections of HCCs and their corresponding peritumour liver tissues of the 31 patients were detected by immunohistochemistry (SABC methods) for protein expression of survivin and MVD, and 10 normal liver tissue sections were as used as negative controls. The correlation among the perfusion parameters, survivin expression, MVD and pathology grade were analysed. RESULTS: (1)The mean values of HAP, HPP, TLP, and HAI of HCCs were 27.50 mL/(min.100 mL), 19.37 mL/(min.100 mL), 46.87 mL/(min.100 mL), and 60.38%, respectively. The mean values of those of the peritumour livers were 14.93 mL/(min.100 mL), 55.70 mL/(min.100 mL), 69.63 mL/(min.100 mL), and 21.51%, respectively. The mean value of those of the normal livers were 12.22 mL/(min.100mL), 74.56 mL/(min.100 mL), 86.78 mL/(min.100 mL), and 14.00%, respectively. The values of HAP and HAI of HCCs were significantly higher than those of the peritumor livers and the normal livers(P<0.01), and the HPP and TLP of HCCs were significant lower than those of the normal livers(P<0.01).The increase of HAP and decrease of HPP of peritumor livers were both significant compared with that of the normal livers(P<0.05). The HAP, HPP, and HAI of HCCs were significantly different from those of peritumor livers (P<0.01)except TLP. (2) Survivin expression in HCCs was detected in 23/31(74.1%), which was significantly higher than that in corresponding non-cancerous adjacent liver tissues and normal liver tissues (P<0.01). Survivin expression was positively correlated with MVD in HCCs. (3) HAP values were significantly and positively correlated with survivin expression (r=0.932,P<0.01)in HCCs.(4)The values of HAP and HAI were correlated with the pathologic grade in HCCs, and those values were increased gradually(P<0.05) among well differentiated HCCs, moderately differentiated, and poorly differentiated HCCs. CONCLUSION CTPI can quantitatively reflect abnormal blood supply of HCCs, which will be helpful for the detection and differentiation of lesions. CT perfusion parameters well correlate with survivin expression, MVD, and the pathologic grade in HCCs, which illustrate that CTPI could hopefully be used to evaluate the angiogenesis and biological behaviors of HCCs prospectively, noninvasively, and dynamically.
Adult ; Aged ; Carcinoma, Hepatocellular ; blood supply ; diagnostic imaging ; metabolism ; pathology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; biosynthesis ; genetics ; Liver Neoplasms ; blood supply ; diagnostic imaging ; metabolism ; pathology ; Male ; Microvessels ; diagnostic imaging ; Middle Aged ; Perfusion Imaging ; methods ; Survivin ; Tomography, Spiral Computed