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1.
AIM: To explore the expression and significance of receptor tyrosine kinase anexelekto (Axl) in nasopharyngeal carcinoma (NPC). METHODS: Immunohistochemistry was used to detect the Axl protein expression of 78 patients with NPC and 32 patients with nasopharyngeal chronic inflammation (NPI). The correlations between the Axl protein levels and the clinical parameters of NPC patients were analyzed. NPC cells were cultured in vitro, and the expression of Axl in well differentiated CNE1 cells, poorly-differentiated CNE2Z cells and undifferentiated C666-1 cells was detected by immunofluorescence staining. After treatment of the CNE1and C666-1 cells with Axl specific inhibitor TP-0903, CCK-8 assay was used to detect cell viability, flow cytometry was adopted to analyze the cell cycle distribution, qPCR was used to examine the mRNA levels of Axl and proliferating cell nuclear antigen (PCNA), and Western blot was used to examine the protein expression of Axl and p-Axl. RESULTS: Axl protein was localized in the cell membrane and cytoplasm. The rate of high expression of Axl in NPC was significantly higher than that in NPI (P<0.01). High Axl expression showed no correlations with NPC patients' age, gender and M stage, while positively correlated with the clinical stage, T stage and N stage (P<0.05). Axl protein showed a low level in the CNE1 cells, but showed a high level in CNE2Z and C666-1 cells. TP-0903 inhibited cell viability in concentration and time dependent manners. TP-0903 at 2 nmol/L showed significant inhibitory effects, as evidenced by arresting the cell cycle at G0 phase and reducing Axl activity and PCNA expression. CONCLUSION: High expression of Axl promotes the clinical progress of NPC.TP-0903 significantly inhibits the viability of NPC cells, suggesting that Axl may be a valuable target in the NPC treatment.  相似文献   

2.
AIM:To observe the expression of calreticulin (CRT) in nasopharyngeal carcinoma tissues, analyze the significance of clinical pathology and the influence on epithelial-mesencymal transition (EMT) of CNE2 cells. METHODS:The expression of calreticulin was detected by immunohistochemistry in 52 nasopharyngeal carcinoma and 57 nasopharyngeal benign tissues, and the significance of clinical pathology was evaluated. The calreticulin gene-specific small interfering RNA was constructed, and then was transfected into the NPC cell line CNE2 using the cationic liposome method. The effect of CRT on the morphological changes of the CNE2 cells was observed under light microscope. The effect of CRT on the cell migration and invasion abilities of the CNE2 cells was detected by Transwell migration and invasion assays. The expression of EMT-related proteins E-cadherin, vimentin, transforming growth factor (TGF)-β and matrix metalloproteinase (MMP)-9 in the CNE2 cells was determined by Western blot. RESULTS:The positive expression rate of CRT in the benign lesion tissues was 19.29% (11/57), which was significantly increased in the nasopharyngeal carcinoma tissues as 82.69% (43/52). The expression rate of CRT was positively correlated with the stage of nasopharyngeal carcinoma and lymph node metastasis (P<0.05). Knockdown of CRT expression made the CNE2 cells showing a spindle shape to a flat, cobblestone-like epithelial state change, arranged more compact, and the migration and invasion abilities were significantly decreased (P<0.05). Knockdown of CRT expression resulted in significant increase in the protein expression of E-cadhe-rin, and the decreases in the protein expression of vimentin, TGF-β and MMP-9 in the CNE2 cells (P<0.05). CONCLUSION:Calreticulin expression in nasopharyngeal carcinoma is significantly higher and positively correlated with nasopharyngeal carcinoma stage and lymph node metastasis. Calreticulin promotes cell migration and invasion of nasopharyngeal carcinoma CNE2 cells by inducing EMT.  相似文献   

3.
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   

4.
AIM: To analyze the effects of special AT-rich sequence binding protein 1 (SATB1) expression on the protein expression profiles in human nasopharyngeal carcinoma (NPC) cells, and to enrich the differential signaling pathways through bioinformatics analysis. METHODS: SATB1 over-expressing lentivirus and negative control lentivirus were used to infect the CNE1 cells, and then the cell lines were obtained by puromycin stressed method. The total proteins of the 2 cells were extracted, and the differentially expressed proteins were screened by TMT-labeled protein quantification technique and tandem mass spectrometry. The mRNA levels of the differential protein-coding genes were verified by RT-qPCR. GO analysis was used to annotate and enrich the differentially expressed proteins, and the KEGG database was used to enrich and analyze the signaling pathways of differential proteins. RESULTS: SATB1 over-expressing CNE1 cells were established through infected with associated lentivirus. Compared with the control group, 278 differentially expressed proteins were identified in SATB1 over-expressing CNE1 cells, in which 115 were up-regulated and 163 were down-regulated. 10 representative differential protein-coding genes were verified by RT-qPCR, which showed the consistence with the proteomic results. GO analysis indicated differentially expressed proteins were mainly involved in cellular processes, single-organism processes, biological regulation, metabolic processes, protein binding and catalysis. Cell components of differentially expressed proteins mainly existed in cell part, cells and organelles. KEGG analysis showed that differentially expressed proteins were involved in signaling pathways closely related to tumors, includeing MAPK, PI3K-Akt, AMPK, JAK-STAT, p53, PPAR, Hippo and HIF-1 signaling pathways. CONCLUSION: Over-expression of SATB1 significantly alters the protein expression profiles in the NPC cells and affects multiple signaling pathways closely related to tumors. Proteomics also provides a possible macro approach to the screening of molecular mechanisms, therapeutic and prognostic targets for NPC.  相似文献   

5.
YU Dan  LIU Xia  FAN Wan-lin  AN Xiang  LI Bing 《园艺学报》2018,34(7):1250-1255
AIM:To investigate the effect of X-ray ionizing radiation on epithelial-mesenchymal transition (EMT) in human nasopharyngeal carcinoma CNE-2 cells and its involved potential signaling pathway. METHODS:The nasopharyngeal carcinoma CNE-2 cells were irradiated with different doses (0 Gy, 2 Gy, 4 Gy and 8 Gy) of X-ray. The morphological changes of the cells were observed under inverted microscope after 24 h. The migration and invasion abilities were detected by wound healing and Transwell assays. The mRNA and protein levels of E-cadherin, N-cadherin and vimentin in nasopharyngeal carcinoma CNE-2 cells were determined by real-time PCR and Western blot, respectively. The protein levels of Akt and p-Akt were detected by Western blot. RESULTS:After X-ray irradiation, the CNE-2 cells exhibited typical ‘cobblestone’ or spindle-like shape, with extended pseudopodia and dilated intercellular space. The invasiveness and metastatic abilities of the CNE-2 cells were enhanced (P<0.01). The mRNA and protein expression levels of E-cadherin were significantly decreased (P<0.01), while the mRNA and protein expression levels of N-cadherin and vimentin were markedly increased after irradiation as compared with the control group (no irradiation) (P<0.05). The protein level of p-Akt was significantly enhanced (P<0.01), while the protein level of Akt showed little change after irradiation. CONCLUSION:X-ray ionizing radiation induces EMT in nasopharyngeal carcinoma CNE-2 cells, which may be related to the activation of PI3K/Akt signaling pathway.  相似文献   

6.
AIM: To investigate the effects of down-regulated miR-9 expression on the proliferation, invasion and migration of nasopharyngeal carcinoma (NPC) cells. METHODS: Human NPC CNE1 and CNE2 cells were transfected with the inhibitor of miR-9 by Lipofectamine to down-regulate the expression of miR-9, and the cells transfected with an inhibitor control were also set up. The cell proliferation and cell cycle were evaluated by CCK-8 assay and flow cytometry. The cell invasion and migration abilities were detected by Transwell invasion and wound-healing assays. Immunoblotting was applied to analyze the levels of the proteins. RESULTS: Compared with control group, inhibition of miR-9 expression in the NPC cells by transfection of the miR-9 inhibitor significantly decreased the proliferation ability (P<0.05). The percentages of the cells in G0/G1 phase ［CNE2: (57.96±1.39)% vs (47.93±1.76)%, P<0.05; CNE1: (51.24±0.88)% vs (48.29±0.39)%, P<0.05］ were significantly increased. The migration distances ［CNE2: (186.50±7.94)μm vs (247.56±15.56)μm, P<0.05; CNE1: (139.06±16.73)μm vs (230.66±14.27)μm, P<0.01］ and the invasion ability of the CNE2 cells (43.00±3.17 vs 65.80±5.20, P<0.01) were also significantly inhibited. Moreover, the tumor cells transfected with the inhibitors produced lower β-catenin. CONCLUSION: Inhibition of miR-9 expression suppresses the proliferation, invasion and migration of nasopharyngeal carcinoma cells.  相似文献   

7.
AIM: To investigate the effects of sphingosine kinase l(SphK1) and focal adhesion kinase(FAK) on the epithelial-mesenchymal transition(EMT) of human colon cancer HCT116 cells. METHODS: Human colon cancer HCT116 cells were divided into 3 groups. N, N-dimethylsphingosine(DMS) was used to suppress the activity of SphK1. PF573228 was used to suppress the activation of FAK. The cells treated with equal volume of culture medium severed as control group. The cell viability was measured by MTT assay. The protein expression of SphK1, FAK and the EMT relative protein E-cadherin, N-cadherin, vimentin and matrix metalloproteinase(MMP) 2 was analyzed by Western blot. The mRNA expression of SphK1, sphingosine-1-phosphate(S1P), FAK, E-cadherin and vimentin was detected by real-time PCR. The ability of tumor cell migration was measured by wound-healing assay. RESULTS: The cell viability of HCT116 cells was suppressed by DMS and PF573228 in dose and time dependent manners. DMS significantly suppressed the expression of SphK1, FAK, N-cadherin, vimentin and MMP2, meanwhile enhanced the expression of E-cadherin. PF573228 reduced the expression of FAK, SphK1, N-cadherin, vimentin and MMP2, meanwhile increased the expression of E-cadherin(P<0.01). In addition, the migration ability of HCT116 cells was significantly decreased by treating with DMS and PF573228(P<0.01). Compared with control group, the mRNA expression of FAK, SphK1, S1P and vimentin was decreased, while the expression of E-cadherin was increased significantly in PF573228 group and DMS group(P<0.05). CONCLUSION: SphK1 and FAK signaling pathways may play an important role in the occurrence of EMT in the colon cancer HCT116 cells.  相似文献   

8.
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   

9.
AIM: To investigate the expression of poly(ADP-ribose) polymerase-1(PARP-1) in the epithelial ovarian cancer(EOC) and its relationship with epithelial-mesenchymal transition(EMT). METHODS: The expression of PARP-1, E-cadherin, vimentin and Snail was detected in the EOC and benign ovarian tumor tissues by immunohistochemical method and real-time PCR. The expression of PARP-1, E-cadherin, vimentin and Snail proteins in the SKOV3 cells treated with efficient PARP-1 inhibitor PJ34 was determined by Western blotting. RESULTS: The positive expression rates of PARP-1, vimentin and Snail were significantly higher in the EOC than that in the benign ovarian tumor tissues, whereas the positive expression rate of E-cadherin was the opposite(P<0.05). The expression of PARP-1, E-cadherin, vimentin and Snail in the EOC was associated with the histological grade, clinical stage and lymphatic metastasis(P<0.05), but no relationship with age and pathological types was observed. The expression of E-cadherin in the EOC was negatively co-related to that of PARP-1. In contrast, the expression of vimentin and Snail in the EOC was positively co-related to that of PARP-1. The relative mRNA expression of PARP-1, vimentin and Snail in the EOC was significantly higher than that in the benign ovarian tumor tissues(P<0.05), while the mRNA expression of E-cadherin in the EOC was remarkably lower than that in the benign ovarian tumor tissues(P<0.05). The protein expression of PARP-1, vimentin and Snail in the SKOV3 cells was significantly decreased(P<0.05), while E-cadherin protein was increased after treated with PJ34(P<0.05). CONCLUSION: PARP-1 may contribute to the onset of EMT in the EOC by regulating the expression of E-cadherin, vimentin and Snail. The role of PARP-1, which is relevant to EMT, might be important in the development of ovarian cancer.  相似文献   

10.
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   

11.
AIM:To investigate the effect of Epstain-Barr virus latent membrane protein 1 (EBV-LMP1) on proliferation and cell cycle of nasopharyngeal carcinoma (NPC) cells. METHODS:The expression of EBV-LMP1 was detected by immunohistochemical method (LSAB). Proliferation of NPC cells was identified by MTT method. Cell cycle percentage was detected by flow cytometry analysis. RESULTS: OD value of EBV-LMP1 expressive NPC cells L-CEN1 was much higher than that of both EBV-LMP1 negative NPC cells V-CEN1 and CNE1 (P＜0.01). Compared with the cell cycle percentage in both V-CNE1 and CNE1, the percentage of G1 was significantly decreased and the percentage of S was much increased in L-CNE1 (P＜0.01). But no obvious differences were observed in all cell cycle percentage between V-CNE1 and CNE1 (P＞0.05).CONCLUSION:The expression of EBV-LMP1 on NPC cell might cause some change of cell cycle and enhance cell proliferation.  相似文献   

12.
SHEN Dong  WANG Wei 《园艺学报》2018,34(9):1622-1626
AIM:To investigate the effects of shikonin on the migration, invasion and epithelial-mesenchymal transition (EMT) in human non-small-cell lung cancer PC9 cells induced by hepatocyte growth factor (HGF). METHODS:The effect of shikonin on the viability of PC9 cells was measured by MTT assay. The cell migration and invasion abilities were analyzed by wound healing assay and Transwell method, respectively. The protein expression levels of E-cadherin, N-cadherin and vimentin in the PC9 cells were determined by Western blot. RESULTS:The viability of PC9 cells was significantly inhibited by shikonin in a dose-dependent manner (P<0.01), with IC50 at 9.364 μmol/L. HGF significantly promoted the abilities of migration and invasion, and induced EMT in the PC9 cells. Shikonin significantly inhibited HGF-induced migration and invasion in the PC9 cells. The expression of E-cadherin was significantly down-regulated and the expression of N-cadherin and vimentin was significantly up-regulated in the presence of HGF (50 μg/L). However, shikonin reversed HGF-induced EMT, as indicated by up-regulation of E-cadherin and down-regulation of N-cadherin and vimentin (P<0.01). CONCLUSION:Shikonin reverses HGF-induced EMT in lung cancer PC9 cells.  相似文献   

13.
LI Bai-he  YUAN Lei 《园艺学报》2016,32(5):852-856
AIM: To investigate the effect of digoxin on hypoxia-induced epithelial-mesenchymal transition (EMT), migration and invasion in human breast carcinoma MCF-7 cells. METHODS: MCF-7 cells were treated in vitro with a chemical hypoxia inducer cobalt chloride (CoCl2) to imitate hypoxia. Cell migration was observed by wound healing assay, and cell invasion was measured by Transwell invasion assay. The protein levels of hypoxia-inducible factor-1α (HIF-1α), Snail, E-cadherin and vimentin in MCF-7 cells were detected by Western blot. RESULTS: Digoxin inhibited CoCl2-induced EMT and reversed the mesenchymal phenotype. CoCl2 enhanced the abilities of migration and invasion (P<0.01), significantly decreased the expression of E-cadherin and increased the expression of HIF-1α, Snail and vimentin (P<0.01), but these effects were blocked by digoxin. CONCLUSION: Digoxin inhibits CoCl2-induced EMT and invasion most likely via HIF1-α-Snail signaling pathway.  相似文献   

14.
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15.
AIM:To investigate the effects of luteolin on the invasion and epithelial-mesenchymal transition (EMT) induced by transforming growth factor-β1 (TGF-β1) in lung cancer A549 cells. METHODS:The effect of luteolin at 5, 10, 20, 40, 80 and 160 μmol/L on the viability of A549 cells was measured by MTT assay. The invasion ability was analyzed by Transwell method. The morphological changes of the A549 cells were observed under microscope.The protein expression of E-cadherin and vimentin in the A549 cells were determined by Western blot. RESULTS:The viability of the A549 cells was significantly inhibited by luteolin in a dose-time dependent manner (P<0.05). The IC50 of luteolin for the A549 cells (24 h) was 68.79 μmol/L, while that (48 h) was 47.86 μmol/L. TGF-β1 induced morphological alteration of the A549 cells from epithelial to mesenchymal forms. Luteolin significantly inhibited TGF-β1-induced invasion of the A549 cells (P<0.01). The protein expression of E-cadherin was significantly down-regulated and the protein expression of vimentin was significantly up-regulated in the presence of TGF-β1 at 5 μg/L (P<0.01). However, luteolin reversed TGF-β1-induced EMT, up-regulation of E-cadherin and down-regulation of vimentin (P<0.01). CONCLUSION:Lu-teolin reverses TGF-β1-induced EMT in the lung cancer A549 cells.  相似文献   

16.
AIM: To investigate the inhibitory effect of microRNA-145 (miR-145) on epithelial-mesenchymal transition (EMT) in renal cancer A-498 cells. METHODS: The A-498 cells were transfected with miR-145 mimics (M145) and mimic negative control(MNC), which served as M145 group and MNC group, respectively. Mock control (MC) group was set up using untreated A-498 cells. The expression level of miR-145 in each group was detected by RT-qPCR. Transwell assay was used to detect the invasion ability of the cells. The protein expression of vimentin, E-cadherin and ADAM28 was determined by Western blot. Bioinformatic method was used to predict the target genes of miR-145. Antagonistic effect of ADAM28 over-expression on the inhibition of EMT by miR-145 was detected by Western blot. The relationship between miR-145 and ADAM28 was analyzed by dual-luciferase reporter assay. RESULTS: The expression level of miR-145 in M145 group was significantly up-regulated than that in MC group (P<0.05). The number of invasive cells in M145 group was 12.78±3.37, which was significantly lower than that in MC group (P<0.05). ADAM28 may be the target gene of miR-145. Compared with MC group, the protein expression of vimentin and ADAM28 in M145 group was significantly decreased (P<0.05), while the protein expression of E-cadherin was significantly increased (P<0.05).After ADAM28 over-expression, the protein expression of vimentin in the A-498 cells of M145 group was significantly increased (P<0.05), and the protein expression of E-cadherin was significantly decreased (P<0.05). The results of dual-lucife-irasei reporter assay showed that ADAM28 was a downstream target gene of miR-145. CONCLUSION: miR-145 may inhibit the expression of EMT-related proteins through the downstream target gene ADAM28 and inhibit the EMT process of renal cancer A-498 cells.  相似文献   

17.
AIM:To investigate the effect of interleukin-6 (IL-6) on epithelial-mesenchymal transition (EMT), migration and invasion of papillary thyroid carcinoma TPC-1 cells by inducing the expression of long noncoding RNA lncTCF7. METHODS:The effects of IL-6 on the expression of lncTCF7 in the TPC-1 cells were detected by RT-qPCR after the TPC-1 cells were treated with IL-6 at 0, 5, 10, 20 and 50 μg/L for 24 h or with IL-6 at 50 μg/L for 0, 6, 12 and 24 h. After the TPC-1 cells were treated with IL-6 at 50 μg/L for 24 h, the effect of IL-6 on the protein expression of E-cadherin and vimentin in the TPC-1 cells was detected by Western blot. The TPC-1 cell line with lncTCF7 over-expression was established, and the effects of lncTCF7 over-expression on EMT, migration and invasion of the TPC-1 cells were measured by Western blot and Transwell assay. After knockdown of lncTCF7 expression and exposure to IL-6 at 50 μg/L, the effects of lncTCF7 on EMT, migration and invasion of TPC-1 cells treated with IL-6 were observed. RESULTS:The expression of lncTCF7 in the TPC-1 cells was induced by IL-6 in a dose-and time-dependent manner. The expression of E-cadherin was down-regulated, the expression of vimentin was up-regulated, and the migration and invasion abilities of the TPC-1 cells were enhanced by lncTCF7 over-expression (P<0.05). The expression of E-cadherin was decreased, the expression of vimentin, Snail and Slug was increased, and the migration and invasion abilities of the TPC-1 cells and intercellular space were enhanced by IL-6. The above changes induced by IL-6 were significantly inhibited by knockdown of lncTCF7 expression. CONCLUSION:IL-6 promotes the EMT, migration and invasion of papillary thyroid carcinoma TPC-1 cells by inducing the expression of lncTCF7.  相似文献   

18.
AIM: To investigate the possible signaling pathway that promotes epithelial-mesenchymal transition (EMT) of the lung cancer A549 cells stimulated with muscarinic receptor 3 (M3R) agonist carbachol. METHODS: The lung cancer cells A549 were treated with 400 μmol/L carbachol. The morphological changes of the cells were observed under inverted phase contrast microscope. The migration and invasion abilites were measured by Wound healing and Transwell assays. qPCR was used to detect the mRNA level of vimentin and E-cadherin. The protein levels of p-AKT, vimentin and E-cadherin were determined by Western blot. RESULTS: After treatment with carbachol, the A549 cells showed loss of the close connection and the cell morphology was transformed from irregular polygon to spindle-like cells. The results of Wound healing and Transwell assays showed that the migration and invasion abilites of the A549 cells were enhanced. Carbachol increased the vimentin expression and decreased the E-cadherin expression at mRNA and protein level (P<0.05). The phosphorylation of AKT in the A549 cells was up-regulated (P<0.05). These changes was inhibited by M3R antagonist 4-DAMP. CONCLUSION: Carbachol promotes EMT in the human lung cancer A549 cells by activating PI3K/AKT signaling pathway.  相似文献   

19.
SUN Jie  FU Li-fang 《园艺学报》2017,33(8):1428-1435
AIM: To explore the expression of Dickkopf-1 (DKK1) in human gastric carcinoma cells, and the influences of DKK1 gene silencing on cell invasion. METHODS: The levels of DKK1 in the human gastric mucosa cell line GES-1 and gastric carcinoma cell lines MKN-45 and SGC-7901 were detected by real-time PCR and Western blot. DKK1 gene was silenced by RNA interference, which was verified by real-time PCR, Western blot and ELISA. The cell invasion ability was determined by Transwell assay, and the cell proliferation was inhibited by mitomycin C. The levels of E-cadherin, N-cadherin, vimentin and β-catenin were determined by real-time PCR and Western blot. RESULTS: The expression of DKK1 was significantly higher in MKN-45 cells and SGC-7901 cells than that in GES-1 cells, indicating that DKK1 expression was obviously increased in gastric carcinoma cells. After successful silencing of DKK1 gene in the MKN-45 cells and SGC-7901 cells, the cell invasion ability was markedly decreased in a time-dependent pattern with increased expression of E-cadherin and decreased expression of N-cadherin and vimentin, indicating that DKK1 silencing dramatically inhibited gastric carcinoma cell invasion and epithelial-mesenchymal transition (EMT). The introduction of exogenous recombinant DKK1 (rDKK1) demonstrated the promoting effect of DKK1 on gastric carcinoma cell invasion and EMT. In addition, the inhibitory effects of DKK1 silencing on gastric carcinoma cell invasion and EMT were fulfilled by down-regulating β-catenin. CONCLUSION: The expression of DKK1 is significantly increased in human gastric carcinoma cells. Silencing of DKK1 markedly inhibits gastric carcinoma cell invasion and EMT by down-regulating β-catenin.  相似文献   

20.
AIM: To investigate the effect of transketolase-like protein 1 (TKTL1) on proliferation of human nasopharyngeal carcinoma cells in vitro. METHODS: The siRNA against TKTL1 mRNA was constructed and transfected into human nasopharyngeal carcinoma cells (CNE cell line). The activity of transketolase was detected before and after RNA interference.Real-time PCR was used to determine the mRNA expression of transketolase (TKT) gene family in the CNE cells.Flow cytometry and MTT test were used to detect the effect of anti-TKTL1 siRNA on cell proliferation and cell cycle in the CNE cells. RESULTS: The total transketolase activity was significantly decreased in the CNE cells transfected with siRNA TKTL1 construct compared with the cells transfected with control vector or untransfected CNE cells. No significant difference in the expression level of TKT and TKTL2 gene between the CNE cells transfected with siRNA TKTL1 construct and the cells transfected with control vector or untransfected CNE cells was observed (P>0.05). However, the expression level of TKTL1 gene was significantly downregulated in the CNE cells transfected with siRNA TKTL1 construct compared with the cells transfected with control vector.Cancer cells were arrested in G0/G1 phase, and cancer cell proliferation was significantly inhibited in the CNE cells transfected with siRNA TKTL1 construct. CONCLUSION: TKTL1 plays an important role in the total transketolase activity and cell proliferation of human nasopharyngeal carcinoma. TKTL1 may be considered as a potential target for novel anti-cancer therapy.  相似文献   

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