共查询到20条相似文献，搜索用时 140 毫秒
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AIM: To establish nasopharyngeal carcinoma(NPC) cell lines stable expressing NPC-derived latent membrane protein 1(LMP1) gene.METHODS:General expression vector and epithelium-specific expression vector of NPC-derived LMP1 gene were constructed by using recombinant techniques, then transfected these vectors into a poor differentiated NPC cell line named CNE-2 ,integration and expression of N-LMP1 in CNE-2 cells were detected by PCR,RT-PCR and Western blot. RESULTS:(1) General expression vector and epithelium-specific expression vector of NPC-derived LMP1 gene were constructed successfully.(2) It showed that N-LMP1 gene expressed in CNE-2 cells correctly.CONCLUSION: The first NPC cell lines which stable express NPC-LMP1 were established. The cell lines obtained will provide important basis for exploring the role of NPC-LMP1 in nasopharynx carcinogenesis. 相似文献
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AIM: To investigate the possible effect of different Latent membrane protein 1 (LMP1) variants on proliferation characteristics of a human well-differentiated nasopharyngeal carcinoma (NPC) cell line CNE1. METHODS: The plasmids which carried EBV-LMP1 gene cloned from B95-8 lymphocyte (B95-8-LMP1) and NPC tissues (CAO-LMP1) were introduced into CNE1 by liposomal transfection. Expression of LMP1 in CNE1 was identified by RT-PCR and Western blot, respectively. Different Effects of the two EBV-LMP1 variants on proliferation characteristics of CNE1 including growth curve, cell cycle distribution and clony-forming efficiency were investigated by means of crystal violet staining proliferation assay, flow cytometry and plastic plate clony-forming assay, respectively. RESULTS: Two transfected cell lines stably expressing different LMP1 variants were established successfully. Either of the two LMP1 variants increased CNE1 growth rate, proliferation index (PI) and clony-forming rate significantly and CAO-LMP1 had more significant growth-promoting effect on CNE1 than B95-8-LMP1. CONCLUSION: It can be concluded from the study that CAO-LMP1 promotes CNE1 proliferation more effectively than B95-8-LMP1. 相似文献
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AIM:To investigate the expression map of two p53 binding proteins 53BP1 and 53BP2 in nasopharyngeal carcinoma (NPC) tissue. METHODS:The expression of 53BP1 and 53BP2 mRNA in NPC biopsy and control group are tested by RT-PCR. The expression of two mRNA in NPC paraffin section are examined by in situ hybridization. RESULTS:No expression of 53BP1 mRNA was found in NPC tissue and control group. However, expression of 53BP2 was detected in NPC biopsy and control group by RT-PCR, specific expressoin found cancerous nest in NPC paraffin section by in situ hybridization. CONCLUSION:The high expression of 53BP2 may be related to the development of NPC. 相似文献
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AIM: To construct the shRNA targeting anterior gradient protein 2 (AGR2) gene for exploring the effect of AGR2 on the biological behavior of nasopharyngeal carcinoma (NPC) cells.METHODS: The expression of AGR2 at mRNA and protein levels in NPC cell lines 6-10B and 5-8F was detected by real-time PCR and Western blot. The pSR-GFP/Neo-AGR2-shRNA expression vector targeting AGR2 was constructed. Based on the interference targeting AGR2, the cell migration and motility were determined by Transwell migration and motility assays.RESULTS: The expression of AGR2 was increased in NPC cell line 5-8F compared with NPC cell line 6-10B (P<0.05). When the AGR2 expression in 5-8F cells was interfered, the cell migration, invasion and tumorigenicity were weakened.CONCLUSION: The expression of AGR2 is up-regulated in NPC cell line 5-8F. pSR-GFP/Neo-CLU-shRNA successfully inhibits the expression of AGR2 in NPC cell line 5-8F. AGR2 inhibits the migration, invasion and tumorigenicity of 5-8F cells in vivo. 相似文献
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AIM: To construct a prokaryotic expression plasmid containing Epstein-Barr viral (EBV) capsid antigen BFRF3 gene and to observe the application of recombinant BFRF3 protein in the serological diagnosis of nasopharyngeal carcinoma (NPC).METHODS: DNA extracted from the B95-8 cells was used as the templates. Polymerase chain reaction (PCR) was used to generate a DNA fragment of BFRF3 gene, and a 531-bp DNA fragment was inserted into a PGEX-5X-1 vector. The recombinant plasmid was transformed into E.coli BL21 (DE3). The expression of GST/BFRF3 fusion protein was induced by IPTG, identified by both SDS-PAGE and Western blotting, and then purified by glutathione-sepharose beads. The purified recombinant protein was coated to microplate for ELISA detection of EBV-IgA antibody in NPC patients.RESULTS: The GST/BFRF3 fusion protein was successfully expressed in E. coli. The molecular weight of the product was approximately 44 kD. The recombinant fusion protein GST/BFRF3 showed good immunoreactivity. A novel ELISA was established using GST/BFRF3 protein. Serum samples collected from the NPC patients and healthy controls were tested by this ELISA. The sensitivity and specificity of GST/BFRF3 tests for NPC patients were 65％ and 87％, respectively.CONCLUSION: The recombinant protein GST/BFRF3 is expressed in E.coli, and it has diagnostic value for screening of NPC patients. 相似文献
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AIM: To explore the effect of EBV infection on growth and apoptosis of nasopharyngeal carcinoma(NPC) cell line.METHODS: NPC cell line CNE1 was directly infected by Epstein Barr virus (EBV). The expression of EBV-latent membrane protein 1 (EBV-LMP1) and bcl-2 were detected by immunohistochemistry method (LSAB). The growth of NPC cells was identified by MTT method. Apoptotic carcinoma cells were detected by flow cytometry analysis and the terminal deoxynucletidyl transferase-medicated dUTP-biotin nick end labeling (TUNEL) methods. RESULTS: EBV-LMP1 was positive in CNE1 infected by EBV(E-CNE1). Compared with CEN1, the growth of E-CNE1 apparently increased (P＜0.01). No apoptotic carcinoma cells were detected and bcl-2 postive cells were 2%~3% respectively in 2 kinds of NPC cells.CONCLUSION: Growth of NPC cells is enhanced by EBV infection and EBV-LMP1 expression, but no influence on expression of bcl-2 and apoptosis of NPC cells. 相似文献
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AIM:To explore the effect of EBV infection on growth and apoptosis of nasopharyngeal carcinoma(NPC)cell line.METHODS:NPC cell line CNE1 was directly infected by Epstein Barr virus(EBV).The expression of EBV-latent membrane protein 1(EBV-LMP1)and bcl-2 were detected by immunohistochemistry method(LSAB).The growth of NPC cells was identified by MTT method.Apoptotic carcinoma cells were detected by flowcytometry analysis and the terminal deoxynucletidyl transferase-medicated dUTP-biotin nick end labeling(TUNEL)methods.RESULTS:EBV-LMP1 was positive in CNE1 infected by EBV(E-CNE1).Compared with CEN1, the growth of E-CNE1 apparently increased(P<0.01).No apoptotic carcinoma cel s were detected and bcl-2 postive cells were 2%~3%respectively in 2 kinds of NPC cells.CONCLUSION:Growth of NPC cells is enhanced by EBV infect ion and EBV-LMP1 expression, but no influence on expression of bcl-2 and apoptosis of NPC cells. 相似文献
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AIM: To study the effect of NF-κB decoy oligodeoxynucleotides(ODNs) on TLR4 and IL-8 expression in LPS-induced SW480 cells. METHODS: SW480 cells were cultured in vitro and stimulated for 3 h with LPS (10 μg/L). NF-κB decoy oligodeoxynucleotides mediated by lipofectin 2000 were added into the cell culture for 6 h. The supernatants were collected and messured for IL-8 by ELISA. TLR4 mRNA and IL-8 mRNA were examined by RT-PCR, respectively. The results were compared with control group, Scrambled ODNs group and lipofectin 2000 group. RESULTS: After SW480 cells were stimulated by LPS, TLR4 mRNA, IL-8 mRNA and IL-8 expressions were significantly increased, and the difference compared with control group was obvious. After treated with NF-κB decoy oligodeoxynucleotides, TLR4 mRNA, IL-8 mRNA and IL-8 expressions were significantly inhibited. The Scrambled ODNs group and lipofectin 2000 group had no effect on them. CONCLUSION: NF-κB decoy ODNs will become a new gene drug for treating inflammatory bowel disease(IBD). 相似文献
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AIM: To observe whether transfection of mammalian expression vector pEGFP containing the gene of B-cell specific moloney leukemia virus insertion site 1(BMI-1) could express in human cervix cancer cell line HeLa, and to detect its effect on HOX family expression and cell cycle.METHODS: pEGFP-BMI-1 was transfected into HeLa cells with Lipofectamine 2000. The expression of pEGFP-BMI-1 was determined by EGFP fluorescence and Western blotting. SYBR green I real-time RT-PCR was used to quantitate mRNA expression of P16INK4a, hTERT, HOXA9, HOXB4 and HOXC13. FACS analysis was used to detect the change of cell cycle.RESULTS: In HeLa cells transfected with pEGFP-BMI-1, the results of real-time RT-PCR showed that the mRNA expressions of P16INK4a, HOXA9 and HOXC13 were reduced to 9.2%, 10.9% and 69.7%, respectively, as compared to control HeLa cells (P<0.01). However, hTERT and HOXB4 mRNA expressions did not change significantly (P>0.05). FACS analysis showed a decrease from 65.68 % to 50.53% in G1 population and a significant increase from 27.17% to 39.59 % in S population after transfection (P<0.01).CONCLUSION: BMI-1 over-expression in HeLa cells down-regulates mRNA expressions of P16INK4a, HOXA9 and HOXC13, decreases G1 population and increases S population. Therefore, BMI-1 may be involved in carcinogenesis and cancer development. 相似文献
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AIM: To investigate whether homocysteine (HCY) induce the expression of macrophage inflammatory protein-1α(MIP-1α)in cultured THP-1 monocytes. METHODS: After exposure of THP-1 monocytes to HCY at increasing concentrations (0.05,0.1 and 0.2 mmol/L) for 8 h, or at 0.1 mmol/L of HCY for different incubation times (4, 8 and 16 h), the expressions of MIP-1α mRNA and protein were determined by RT-PCR and immunocytochemistry, respectively. RESULTS: RT-PCR showed that the expression of MIP-1α mRNA increased with the concentrations of HCY compared with the control group. Meanwhile, after the treatment of 0.1 mmol/L HCY to the cells for different times, the MIP-1α mRNA expression increased at 4 h, peaked at 8 h, and then decreased at 16 h. The authenticity of RT-PCR products was confirmed by DNA sequencing. Image analysis of Immunocytochemistry assay showed the expression of MIP-1α protein in experimental groups increased in a dose- and time-dependent manner(P＜0.01). CONCLUSIONS: HCY induced monocytes to express MIP-1α mRNA and protein. 相似文献
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AIM: To observe the effect of B7-H4 gene transfection on human ovarian cancer cell growth and its tumor formation. METHODS: Human ovarian cancer cell line SKOV3 was transfected with PEGFP-N1/B7-H4 and PEGFP-N1 by LipofectamineTM2000 method. The expression of B7-H4 gene was detected by RT-PCR. The high expression cell strain was selected. The growth curve was drawn by MTT methods. The tumor size was observed after SKOV3/B7-H4 cells, SKOV3/neo cells and SKOV3 cells were injected subcutaneously into SCID mouse. RESULTS: The expressions of B7-H4 mRNA and fusion protein in SKOV3/B7-H4 cells were positive. Compared with the other two groups, transfection significantly promoted cell proliferation in vitro. In addition, facilitated tumor formation and enhanced tumor growth were observed in SKOV3/B7-H4 group. No difference in tumor growth between SKOV3 group and SKOV3/N1 group was observed. CONCLUSION: B7-H4 may be a candidate gene for gene therapy of ovarian cancer. 相似文献
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XI Shao-yan WU Qiu-liang ZONG Yong-sheng YONG Juan-juan LIANG Jian-zhong HOU Jing-hui LU Jia-bin 《园艺学报》2012,28(4):643-648
AIM: To investigate the protein expression of 2 receptor tyrosine kinases,epidermal growth factor receptor(EGFR) and c-Kit, in non-keratinizing nasopharyngeal carcinoma (NPC) and to analyze the relationship of that with the clinicopathological parameters. METHODS: Ninety-five samples of stage II and III non-keratinizing NPC biopsies were collected from the Department of Pathology, Sun Yat-sen University Cancer Center. The 10% formalin-fixed paraffin-embedded biopsy blocks were re-sectioned. Besides HE routine staining, immunohistochemistry was performed for detecting the expression of EGFR, c-Kit, latent membrane protein 1(LMP1) and Ki-67. RESULTS: The protein expression rates of EGFR and c-Kit were 70.53% (67/95) and 63.16% (60/95), respectively. The expression score of EGFR was positively correlated with that of c-Kit protein (P<0.05). They were both correlated with T staging (EGFR,P<0.05; c-Kit, P<0.01). Furthermore, the staining intensities of EGFR and c-Kit proteins were also correlated with T staging(EGFR, P<0.05; c-Kit,P<0.01). CONCLUSION: The proteins of EGFR and c-Kit are usually expressed in non-keratinizing NPC cells. The immunoreactive scores of these 2 receptor tyrosine kinases are positively correlated with each other. Either the expression rate or the immunoreactive intensity of EGFR and c-Kit proteins is correlated with primary tumor progression. Immunohistochemical staining of EGFR or c-Kit protein in NPC biopsies could be recognized as an insight into further gene analysis for target therapy. 相似文献
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WANG Si-si LIAO Xiao-min SHAO Zhong-ming YUAN Jian-ling FENG Mu-yin HA Yan-ping LI Ru-jia SHEN Zhi-hua JIE Wei 《园艺学报》2018,34(12):2172-2179
AIM:To establish SETD2 gene knockout nasopharyngeal carcinoma (NPC) cell strains based on CRIPSR/Cas9 technique and to analyze their proliferation characteristics. METHODS:Sub-quantitative RT-PCR and Western blot were used to detect the expression of SETD2 in immortalized nasopharyngeal epithelial cell line NP-69, well differentiated NPC cell line CNE1, poorly-differentiated NPC cell line CNE2Z and undifferentiated NPC cell line C666-1, and the SETD2 high expression cell line CNE1 was screened. The proliferation ability of CNE1 cells before and after the SETD2 gene knockout was analyzed by CCK-8 and colony formation assay. The cell cycle distribution was detected by flow cytometry, and the expression of cell cycle-related proteins was detected by Western blot. RESULTS:Compared with NP-69 cells, the expression of SETD2 was decreased gradually in CNE1, CNE2Z and C666-1 cells (P<0.01). Based on the CRISPR/Cas9 technique, 2 monoclonal cell strains with SETD2 gene stable knockout, named CNE1-SETD2-KO-#5 and #9, were successfully screened from total 15 monoclones. The results of CCK-8 and plate colony formation assay confirmed that the proliferation ability of CNE1-SETD2-KO-#5 and #9 cells was significantly enhanced compared with CNE1-WT cells (P<0.05). The results of flow cytometry analysis showed that the G1 phase of CNE1-SETD2-KO-#5 and #9 cells was decreased, while the G2/M and S phases were increased significantly (P<0.05). The results of Western blot confirmed the increases in the protein levels of proliferating cell nuclear antigen (PCNA), cyclin D1, cyclin B1, cyclin A2, cyclin E1, cyclin-dependent kinases 2 (CDK2) and CDK4, and the decrease in the protein level of p21 after SETD2 gene knockout (P<0.05). CONCLUSION:The NPC cell strains with SETD2 gene knockout were successfully constructed based on CRISPR/Cas9 technique. SETD2 expression correlates with cell differentiation status in the NPC cells. SETD2 gene knockout promotes NPC cell proliferation by up-regulating cyclin D1, cyclin B1, cyclin A2, cyclin E1, CDK2 and CDK4, and down-regulating p21 expression. 相似文献
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AIM: The present study was undertaken to investigate the effect of angiotensin II (AngⅡ) on expression of MMP-9 in THP-1 macrophages. METHODS: Macrophages converted from THP-1 monocytes by incubating with PMA (0.1 μmol/L) for 48 h were divided into PMA group; PMA+AngⅡ group (10-7mol/L, 1 h); PMA+AngⅡ+PDTC group (10 μmol/L, 30 min) and PDTC group. Western blotting was used to detect the MMP-9 and phosphorylation of NF-κB p65, and the expression of MMP-9 mRNA in THP-1 macrophages was measured by RT-PCR.RESULTS: Compared to control group, the expression of MMP-9 (1.06±0.11, P<0.05) and phosphorylation of NF-κB p65 (1.02±0.10, P<0.05) in THP-1 macrophages were expressed when treated with AngⅡ (10-7mol/L); and the expression of MMP-9 mRNA were upregulated (1.22±0.08, P<0.05). However, NF-κB inhibitor PDTC reduced the NF-κB p65 (0.99±0.12, P<0.01) and MMP-9 (1.04±0.14, P<0.01) expressions and decreased the expression of MMP-9 mRNA (0.90±0.06，P<0.01). CONCLUSION: NF-κB signaling pathway contributes to the expression of MMP-9 in THP-1 macrophage induced by AngⅡ. 相似文献
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AIM: To study the signal pathway involved in up-regulation of LPS-induced HO-1 expression by CCK-8. METHODS: Forty-two SD rats were divided into 7 groups (six rats each) randomly as follows: control group, LPS group, LPS+SP600125 (JNK-specific inhibitor) group, CCK-8+LPS group, CCK-8+LPS+SP600125 group, CCK-8 group and CCK-8 +SP600125 group. Lungs from the rats in these 7 groups were excised 6 h after the agents were administered. HO-1 mRNA expression was examined by RT-PCR. The protein expression of HO-1 was detected by Western blotting and immunofluorescence flow cytometry (FCM). RESULTS: There were significant positive expression of HO-1 mRNA in LPS group compared to control group. CCK-8 enhanced LPS-induced HO-1 mRNA expression and CCK-8 alone induced HO-1 mRNA expression as well. The mRNA expressions of HO-1 in LPS group, CCK-8+LPS group and CCK-8 group were 3.01 (P<0.01), 5.88 (P<0.01) and 3.45 (P<0.01) times as many as that in control group, respectively. SP600125 inhibited the mRNA expression of HO-1 induced by CCK-8 and (or) LPS. The change of HO-1 protein expression was in accordance with that of HO-1 mRNA expression by Western blotting and immunofluorescence FCM. CONCLUSION: These results suggest that JNK/c-Jun signal pathway plays an important role in the up-regulation of LPS-induced HO-1 expression by CCK-8. 相似文献