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Objective:To investigate the feasibility of dynamic contrast-enhanced (DCE)-MRI in assessing the depth of invasion (DOI) of early tongue squamous cell carcinoma.Methods:From January 2016 to December 2020, a total of 100 patients with early tongue squamous cell carcinoma confirmed by postoperative pathology were retrospectively analyzed in the Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine. The study included 48 cases of T1 stage and 52 cases of T2 stage. All patients underwent routine MRI, DCE-MRI and contrast-enhanced T 1WI (CE-T 1WI) before surgery. The DOI was measured on images at different phases of axial DCE-MRI (30, 60 and 120 s after contrast injection) and CE-T 1WI (MRI-DOI) by 2 doctors independently. The intraclass correlation coefficient (ICC) was applied to evaluate the consistency of the measurements. MRI-DOI measured on DCE-MRI 30, 60, 120 s, CE-T 1WI and pathological DOI measured on biopsies were statistically analyzed with analysis of variance for repeated measurement and least significant difference t test. Pearson correlation coefficient was used to compare the correlation between MRI-DOI and pathological DOI. Receiver operation characteristic (ROC) curve analysis was used to explore the performance of MRI-DOI for clinical T1 and T2 staging. Results:There was a good consistency in MRI-DOI measured on DCE-MRI 30, 60, 120 s, and CE-T 1WI images with ICC of 0.752, 0.875, 0.883, and 0.841, respectively. The values of MRI-DOI were (8.35±3.52), (6.88±2.41), (7.52±2.65) and (8.60±3.39) mm, respectively, and pathological DOI was (5.75±2.01) mm. There was statistically significant difference in the overall comparison among different phases of MRI-DOI and pathological DOI ( F=69.25, P<0.001). MRI-DOI were significantly higher than pathological DOI ( P<0.05). All MRI-DOI measured on DCE-MRI 30, 60, 120 s and CE-T 1WI correlated positively with pathological DOI ( r=0.574, 0.851, 0.731, 0.663, all P<0.001). MRI-DOI derived from DCE-MRI 60 s showed the highest diagnostic efficiency for T1 and T2 staging (area under the ROC curve was 0.931, 95%CI 0.881-0.982). When the optimal cutoff value was 6.0 mm, the accuracy, sensitivity and specificity were 88.0%, 96.2% and 79.2%, respectively. Conclusions:DCE-MRI can be applied to assess DOI in early tongue squamous cell carcinoma. MRI-DOI based on DCE-MRI 60 s has the best correlation with pathological DOI and has a potential to predict clinical T staging in early tongue squamous cell carcinoma.
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Objective:To explore the expression of vasohibin-1 (VASH1) and vascular endothelial growth factor A (VEGF-A) and clinical significance of VASH1 in gastric carcinoma.Methods:The expression of VASH1 and VEGF-A were detected by immunohistochemistry in formalin-fixed and paraffin-embedded sections in 56 pairs of gastric cancer and corresponding paraneoplastic tissue specimens. The correlation between the expression of VASH1, VEGF-A, clinicopathological parameters and prognosis were analyzed.Results:VASH1 and VEGF-A expression was significantly higher in gastric cancer tissues than normal paraneoplastic tissues. VASH1 and VEGF-A protein were expressed in 79% and 82% of gastric cancer tissues, respectively. A positive correlation was found between Vasohibin-1 and VEGF-A expression in gastric cancer tissues. VASH1 expression has significant positive correlation with TNM stage, tumor stromal invasion, tumor gross types and distant metastasis. Patients with high VASH1 expression had significantly worse overall survival (OS) and progression-free survival (PFS) than those with low VASH1 expression.Conclusion:VASH1 might be a clinically relevant predictor of patients in gastric cancer.
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Objective:To investigate the effects of long non-coding RNA (lncRNA) FBXL19-AS1 on the proliferation, migration and invasion of pancreatic cancer cells, and to determine the targeting relationship of lncRNA FBXL19-AS1 and microRNA-339-3p (miR-339-3p).Methods:From January 2017 to August 2019, 73 cancer tissues and matched normal pancreatic tissues adjacent to cancer from patients pathologically diagnosed as pancreatic cancer who underwent surgical resection in Yantai Hospital of Yantai were collected. Normal pancreatic epithelial cells (hTERT-HPNE) and 3 pancreatic cancer cell lines (Capan-1, SW1990, PaTu8988) were cultured in vitro. The real-time fluorescent quantitative PCR was used to detect the expression of lncRNA FBXL19-AS1 and miR-339-3p in pancreatic cancer tissues and cell lines. The Capan-1 cells were divided into the NC group (normal control group), si-NC group (transfected with meaningless negative sequence), si-FBXL19-AS1 group (transfected with FBXL19-AS1 small interfering RNA), miR-NC group (transfected with empty plasmid control), miR-339-3p group (transfected with miR-339-3p overexpression lentiviral vector), si-FBXL19-AS1+ anti-miR-339-3p NC group (cotransfected with FBXL19-AS1siRNA and miR-339-3p inhibitor negative control sequence) and si-FBXL19-AS1+ anti-miR-339-3p group (cotransfected with FBXL19-AS1siRNA and miR-339-3p inhibitor). CCK-8 method was used to detect cell proliferation activity. Transwell chamber was used to detect cell migration and invasion ability, and western blotting method was used to detect cell cyclinD1, matrix metalloproteinase 2 (MMP2) and MMP9 protein expression. Bioinformatics and dual luciferase reporter gene experiments were used to analyze the targeting relationship between lncRNA FBXL19-AS1 and miR-339-3p.Results:The expression of lncRNA FBXL19-AS1 in pancreatic cancer tissue was significantly higher than that in normal pancreatic tissue adjacent to cancer (2.96±0.21 vs 1.00±0.13, P<0.05), and the expression of miR-339-3p was significantly lower than that in normal pancreatic tissue adjacent to cancer (0.37±0.05 vs 1.00±0.11, P<0.05). The expression of lncRNA FBXL19-AS1 in Capan-1, SW1990 and PaTu8988 cells were 2.43±0.18, 1.97±0.13 and 1.73±0.14, respectively, which were significantly higher than that of hTERT-HPNE cells 1.00±0.07. The expression of miR-339-3p were 0.42±0.03, 0.54±0.03 and 0.57±0.04, respectively, which were all significantly lower than 1.00±0.05 of hTERT-HPNE cells. Among them, the expression of lncRNA FBXL 19-AS1 in Capan-1 cells was the highest, and the miR-339-3p was the lowest. Compared with the si-NC group, the absorbance value ( A450) of Capan-1 cells in the si-FBXL19-AS1 group, the number of migrating cells, and the number of transmembrane cells were significantly decreased (0.47±0.03 vs 0.94±0.06, 81.00±7.41 vs 187.00±16.13, 67.00±5.41 vs 141.00±9.24), the protein expression of cyclinD1, MMP2 and MMP9 was significantly reduced (0.44±0.03 vs 0.83±0.05, 0.48±0.03 vs 0.92±0.07, 0.38±0.02 vs 0.73±0.05). Compared with the miR-NC group, the A450, the number of migrating cells, and the number of transmembrane cells of Capan-1 cells in the miR-339-3p group were significantly decreased (0.54±0.04 vs 0.94±0.05, 98.00±6.53 vs 193.00±10.02, 83.00±6.58 vs 146.00±7.11, the protein expression of cyclinD1, MMP2 and MMP9 was significantly reduced (0.47±0.03 vs 0.81±0.07, 0.43±0.03 vs 0.94±0.06, 0.32±0.02 vs 0.71±0.06). Compared with the si-FBXL19-AS1+ anti-miR-NC group, the A450, the number of migrating cells and the number of transmembrane cells in the si-FBXL19-AS1+ anti-miR-339-3p group increased significantly (0.96±0.07 vs 0.48±0.03, 204.00±11.25 vs 83.00±5.11, 157.00±8.76 vs 64.00±4.12, P all <0.05), the protein expression of cyclinD1, MMP2 and MMP9 increased significantly (0.84±0.06 vs 0.42±0.03, 0.96±0.08 vs 0.47±0.08, 0.74±0.06 vs 0.37±0.02, P all <0.05). The luciferase activity of Capan-1 cells cotransfected with WT-FBXL19-AS1 and miR-339-3p mimics was significantly lower than that of the cotransfected with WT-FBXL19-AS1 and miR-NC (0.47±0.04 vs 1.00±0.10, P all <0.05). The difference of the luciferase of Capan-1 cells in the cotransfected MUT-FBXL19-AS1 and miR-339-3p mimics group and the cotransfected MUT-FBXL19-AS1 and miR-NC group was not statistically significant. Conclusions:LncRNA FBXL19-AS1 was highly expressed in pancreatic cancer tissues and Capan-1 pancreatic cancer cell lines. Knockdown of lncRNA FBXL19-AS1 can target miR-339-3p to regulate the proliferation, migration and invasion of pancreatic cancer cells, and promote the occurrence and development of pancreatic cancer.
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ABSTRACT Background: Neuroblastoma is a pediatric tumor with a mortality rate of 40% in the most aggressive cases. Tumor microenvironment components as immune cells contribute to the tumor progression; thereby, the modulation of immune cells to a pro-inflammatory and antitumoral profile could potentialize the immunotherapy, a suggested approach for high-risk patients. Preview studies showed the antitumoral potential of BJcuL, a C- type lectin isolated from Bothrops jararacussu venom. It was able to induce immunomodulatory responses, promoting the rolling and adhesion of leukocytes and the activation of neutrophils. Methods: SK-N-SH cells were incubated with conditioned media (CM) obtained during the treatment of neutrophils with BJcuL and fMLP, a bacteria-derived peptide highly effective for activating neutrophil functions. Then we evaluated the effect of the same stimulation on the co-cultivation of neutrophils and SK-N-SH cells. Tumor cells were tested for viability, migration, and invasion potential. Results: In the viability assay, only neutrophils treated with BJcuL (24 h) and cultivated with SK-N-SH were cytotoxic. Migration of tumor cells decreased when incubated directly (p 0.001) or indirectly (p 0.005) with untreated neutrophils. When invasion potential was evaluated, neutrophils incubated with BJcuL reduced the total number of colonies of SK-N-SH cells following co-cultivation for 24 h (p 0.005). Treatment with CM resulted in decreased anchorage-free survival following 24 h of treatment (p 0.001). Conclusion: Data demonstrated that SK-N-SH cells maintain their migratory potential in the face of neutrophil modulation by BJcuL, but their invasive capacity was significantly reduced.
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Neuroblastoma is a pediatric tumor with a mortality rate of 40% in the most aggressive cases. Tumor microenvironment components as immune cells contribute to the tumor progression; thereby, the modulation of immune cells to a pro-inflammatory and antitumoral profile could potentialize the immunotherapy, a suggested approach for high-risk patients. Preview studies showed the antitumoral potential of BJcuL, a C- type lectin isolated from Bothrops jararacussu venom. It was able to induce immunomodulatory responses, promoting the rolling and adhesion of leukocytes and the activation of neutrophils. Methods: SK-N-SH cells were incubated with conditioned media (CM) obtained during the treatment of neutrophils with BJcuL and fMLP, a bacteria-derived peptide highly effective for activating neutrophil functions. Then we evaluated the effect of the same stimulation on the co-cultivation of neutrophils and SK-N-SH cells. Tumor cells were tested for viability, migration, and invasion potential. Results: In the viability assay, only neutrophils treated with BJcuL (24 h) and cultivated with SK-N-SH were cytotoxic. Migration of tumor cells decreased when incubated directly (p < 0.001) or indirectly (p < 0.005) with untreated neutrophils. When invasion potential was evaluated, neutrophils incubated with BJcuL reduced the total number of colonies of SK-N-SH cells following co-cultivation for 24 h (p < 0.005). Treatment with CM resulted in decreased anchorage-free survival following 24 h of treatment (p < 0.001). Conclusion: Data demonstrated that SK-N-SH cells maintain their migratory potential in the face of neutrophil modulation by BJcuL, but their invasive capacity was significantly reduced.(AU)
Subject(s)
Animals , Peptides , Bothrops , Crotalid Venoms/isolation & purification , Lectins, C-Type/isolation & purification , Neuroblastoma , Neutrophils , In Vitro TechniquesABSTRACT
OBJECTIVE: The present study is to evaluate the biological functions of long non-coding RNA (lncRNA), X-inactive specific transcript, X-inactive specific transcript (XIST) in human epithelial ovarian cancer (EOC). METHODS: XIST was upregulated in EOC cell lines, CAOV3 and OVCAR3 cells by lentiviral transduction. The effects of XIST overexpression on cancer cell proliferation, invasion, chemosensitivity and in vivo tumor growth were investigated, respectively. Possible sponging interaction between XIST and human microRNA hsa-miR-214-3p was further evaluated. Furthermore, hsa-miR-214-3p was overexpressed in XIST-upregulated CAOV3 and OVCAR3 cells to evaluate its effect on XIST-mediated EOC regulation. RESULTS: Lentivirus-mediated XIST upregulation had significant anticancer effects in CAOV3 and OVCAR3 cells by suppressing cancer cell proliferation, invasion, increasing cisplatin chemosensitivity and inhibiting in vivo tumor growth. Hsa-miR-214-3p was confirmed to directly bind XIST, and inversely downregulated in XIST-upregulated EOC cells. In EOC cells with XIST upregulation, secondary lentiviral transduction successfully upregulated hsa-miR-214-3p expression. Subsequently, hsa-miR-214-3p upregulation functionally reversed the anticancer effects of XIST-upregulation in EOC. CONCLUSION: Upregulation of lncRNA XIST may suppress EOC development, possibly through sponging effect to induce hsa-miR-214-3p downregulation.
Subject(s)
Humans , Cell Line , Cell Proliferation , Cisplatin , Down-Regulation , MicroRNAs , Neoplasm Invasiveness , Ovarian Neoplasms , RNA, Long Noncoding , Up-RegulationABSTRACT
AIM:To investigate the possible mechanism of microRNA-106a promoting the invasion of human breast cancer MDA-MB-231 cells. METHODS:The efficiencies of transfection with microRNA-106a inhibitor and mi-croRNA-106a mimic by liposome were detected by qPCR. The mRNA and protein expression levels of tissue inhibitor of metalloproteinase 2 (TIMP-2), matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP9) in the MDA-MB-231 cells transfected with microRNA-106a mimic were detected by qPCR and Western blot. The effect of microRNA-106a on the invasion ability of MDA-MB-231 cells was measured by Transwell assay. The luciferase reporter assay was used to detect the regulatory effect of microRNA-106a on the TIMP-2 pathway. RESULTS:In the MDA-MB-231 cells, the ex-pression level of microRNA-106a decreased at 48 h after transfection with microRNA-106a inhibitor (P<0.05), and the expression level of microRNA-106a increased at 48 h after transfection with microRNA-106a mimic (P<0.05). The mi-croRNA-106a inhibitor decreased the invasion ability of MDA-MB-231 cells in vitro (P<0.05). The microRNA-106a mim-ic down-regulated the expression of TIMP-2 and up-regulated the expression of MMP2 and MMP9 (P<0.05) in the MDA-MB-231 cells. The microRNA-106a inhibitor enhanced the luciferase activity of the reporter plasmids containing the 3'-un-translated region of TIMP-2 gene (P<0.05), while the microRNA-106a mimic decreased the luciferase activity of the re-porter plasmid (P<0.05). CONCLUSION:High expression of microRNA-106a promotes the invasion ability of breast cancer MDA-MB-231 cells in vitro, which may be related to the inhibition of TIMP-2 pathway. MicroRNA-106a plays an important role in the invasion of breast cancer MDA-MB-231 cells.
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AIM:To investigate the function of microRNA-125a-5p (miR-125a-5p) on epithelial-mesenchy-mal transition ( EMT) of breast cancer cells via GSK-3β/Snail signaling pathway. METHODS:The expression of miR-125a-5p in normal breast epithelial cells and breast cancer cells, as well as the transfection efficiency of miR-125a-5p plas-mid in MDA-MB-231 cells was detected by RT-qPCR. The chemotaxis ability and invasion ability were detected by chemo-taxis assay and Transwell invasion assay. The changes of EMT-related markers, the protein level of phosphorylated glycogen synthase kinase-3β (p-GSK-3β) and the nuclear translocation of Snail were determined by Western blot. RESULTS:The expression of miR-125a-5p in the breast cancer cells was significantly lower than that in the normal breast epithelial cells. The expression of miR-125a-5p was significantly higher in MDA-MB-231/miR-125a-5p cells than that in MDA-MB-231/NC cells. The ability of epithelial growth factor (EGF) at 10 μg/L to induce chemotaxis of MDA-MB-231 cells was the stron-gest. Compared with MDA-MB-231/NC group, stimulation of EGF decreased the invasion ability of MDA-MB-231/miR-125a-5p cells, and resulted in the increase in E-cadherin expression, while significantly decreased the protein levels of vi-mentin and p-GSK-3β. Meanwhile, the nuclear localization of Snail was significantly inhibited. The invasion capacity of MDA-MB-231/miR-125a-5p+GAB2 cells was significantly enhanced compared with MDA-MB-231/miR-125a-5p +Con cells, the expression of E-cadherin was decreased, and the protein levels of vimentin and p-GSK-3β were significantly in-creased, while the nuclear localization of Snail was promoted. CONCLUSION:miR-125a-5p suppresses EMT via GSK-3β/Snail signaling pathway, thus inhibiting the invasion ability of breast cancer cells.
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AIM:To observe the expression of long noncoding RNA TTTY 15 in osteosarcoma tissues and cell lines and to explore its effect on the viability and invasion ability of osteosarcoma cell lines.METHODS:qPCR was used to detect the expression of TTTY15 in 11 cases of osteosarcoma and its adjacent tissues.The mRNA levels of TTTY15 in os-teosarcoma cell lines(143B,Saos2,MG-63,U2OS and HOS)and human osteoblast cell line hFOB1.19 were also tested. TTTY15 was down-regulated after transfected with small interfering RNA in MG-63 cells,the cell line with the highest level of TTTY15.The effect of TTTY15 knockdown on the viability of MG-63 cells was measured by CCK-8 assay.The cell cycle distribution was analyzed by flow cytometry.The effect of TTTY15 knockdown on the cell invasion ability was detected by Transwell assay.The levels of miR-216b-5p and FOXM1 mRNA were detected by qPCR, and the changes of the related proteins were determined by Western blot.RESULTS:Compared with the adjacent tissues,the expression of TTTY15 in-creased in the osteosarcoma tissues(P<0.01).Compared with the human osteoblast cell line,the expression of TTTY15 increased in the osteosarcoma cell lines(P<0.05), and the level of TTTY15 in the MG-63 cells was the highest(P<0.01).After knockdown of TTTY15 expression in the MG-63 cells,the cell viability was decreased(P<0.05),cell cycle progression was inhibited(P<0.01), and the cell invasion ability was decreased(P<0.01).The expression of miR-216b-5p was increased(P<0.01)and the expression of FOXM1 mRNA was decreased(P<0.01).The protein expres-sion of FOXM1,CDK4,cyclin D1,MMP-2 and N-cadherin was decreased,while the protein expression of E-cadherin was increased(P <0.05).CONCLUSION: The expression of TTTY15 is increased in the osteosarcoma tissues and cell lines.The low expression of TTTY15 inhibits the cell viability and invasion ability of osteosarcoma cells.The possible mechanism is that the knockdown of TTTY 15 expression results in the increase in miR-216b-5p expression and the down-regulation of FOXM1 expression.
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AIM:To investigate the effect of sinomenine on the viability, migration and invasion of human ovarian cancer SKOV3 cells and its possible mechanism.METHODS:The SKOV3 cells were treated with sinomenine at different concentrations for 12 h,24 h and 48 h.CCK-8 assay was employed to detect the effects of sinomenine on the via-bility of the SKOV3 cells.Flow cytometry was used to analyze the cell cycle distribution.The cell migration and invasion abilities were measured by Transwell assay.Western blot was used to determine the protein levels of cyclin A,cyclin D1, E-cadherin and matrix metalloproteinase-9(MMP-9).RESULTS: Sinomenine remarkably inhibited the viability of SK-OV3 cells and IOSE80 cells in a time-dependent and dose-dependent manner(P<0.05),and the IC50values of 48 h were 2.12 mmol/L and 17.35 mmol/L,respectively.In a dose-dependent manner,sinomenine induced G0/G1and S phase ar-rest in SKOV3 cells(P<0.05),suppressed the migration and invasion abilities of SKOV 3 cells(P<0.05),down-regu-lated the protein levels of cyclin A,cyclin D1 and MMP-9(P<0.05), and up-regulated the protein level of E-cadherin (P<0.05).CONCLUSION:Sinomenine inhibits the viability,migration and invasion of human ovarian cancer SKOV 3 cells most likely via down-regulation of the protein levels of cyclin A,cyclin D1 and MMP-9,and up-regulation of the pro-tein level of E-cadherin.
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AIM:To investigate the effect of propofol on the viability,invasion ability and apoptosis of colorec-tal cancer cells.METHODS:Propofol at 10,25,50 and 100 μmol/L was used to treat LoVo cells for 72 h,and propofol at 100 μmol/L was used to treat the LoVo cells for 12,24,48 and 72 h.The cell viability was measured by CCK-8 assay. The invasion ability of the LoVo cells treated with propofol at 100 μmol/L for 72 h was detected by Transwell assay.The cell cycle distribution and cell apoptotic rate were analyzed by flow cytometry.The protein levels of matrix metalloproteinase (MMP)-2,MMP-9,cleaved caspase-3,Notch1 and hairy and enhancer of split 1(Hes1)were determined by Western blot.RESULTS:Propofol inhibited LoVo cell viability.The cell invasion ability,S stage cells,and the protein levels of MMP-2,MMP-9,Notch1 and Hes1 in propofol group were significantly lower than those in control group,and the apoptotic rate,G0/G1cells and the protein level of cleaved caspase-3 were significantly higher than those in control group(P<0.01).CONCLUSION:Propofol inhibits the viability and invasion ability of colorectal cancer LoVo cells,blocks cell cy-cle and induces apoptosis.The mechanism is related to down-regulation of Notch1 signaling pathway.
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AIM:To investigate the expression of long non-coding RNA PVT1 in ovarian cancer and the role of PVT1 in migration and invasion abilities of ovarian cancer cells.METHODS: The expression of PVT1 in ovarian cancer tissue,normal ovarian tissue and different ovarian cancer cell lines was detected by qPCR.Transwell assay was used to de-tect the invasion ability of ovarian cancer cells after PVT1 silencing.The migration ability of the ovarian cancer cells after PVT1 silencing was detected by scratch test.The interaction between PVT1 and microRNA(miR)-551 was analyzed by dual-luciferase reporter assay.The effect of miR-551-inhibitor on the invasion and migration abilities of ovarian cancer cells after PVT1 silencing was detected by Transwell assay and scratch test.The expression of Wnt signaling pathway-related pro-teins was determined by Western blot after PVT1 silencing.The effects of PVT1 silencing on tumor weight and volume of ovarian cancer were examined by subcutaneous tumor transplantation in nude mice.RESULTS:The expression of PVT1 in ovarian cancer tissue was significantly higher than that in normal ovarian tissue(P<0.05).The expression level of PVT1 in ovarian cancer cell line ES-2 was the highest.PVT1 silencing inhibited the invasion and migration abilities of the ovarian cancer cells.After PVT1 silencing,miR-551-inhibitor promoted the invasion and migration abilities of the ovarian cancer cells.The expression of Wnt signaling pathway-related proteins was decreased after PVT1 silencing(P<0.05).Compared with negative control group,the tumor volume and weight in PVT1-siRNA group were significantly decreased(P<0.05). CONCLUSION:PVT1 plays an important role in the development of ovarian cancer.PVT1 regulates the invasion and mi-gration abilities of ovarian cancer cells through Wnt signaling pathway.
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Objective@#To analyze the clinical value of SPECT/CT in diagnosis of skull base bone invasion and clinical decision-making for nasopharyngeal carcinoma (NPC), and to compare their diagnostic value with SPECT/CT, CT, MRI, and MRI combined with SPECT (MRI-SPECT) for skull base bone invasion.@*Methods@#Before treatment, among 348 newly diagnosed NPC patients, CT scan was performed in 186 patients (group A) and the remaining 162 patients received MRI scan (group B). Clinical doctors then made clinical management decisions according to the CT or MRI results. After that, all patients underwent 99Tcm-MDP SPECT/CT examination for nasopharyngeal local tomography, and the results were provided to the clinical doctors to make clinical management decisions again. The changes between the two clinical management decisions were scored according to diagnosis, range of lesion, staging, treatment regimens, and auxiliary examination. The diagnostic value of CT scan, MRI scan, SPECT/CT and MRI-SPECT for skull base bone invasion was then evaluated and compared.@*Results@#In terms of changes in scores of clinical management decisions, the score of group A was 1.387 and group B was 0.951, showing a significant difference between the two groups by Wilcoxon test (Z=6.570, P<0.001). By χ2 test, there were correlations between CT and SPECT/CT (χ2 =98.495, P<0.001), and between MRI and SPECT/CT (χ2 =32.662, P<0.001). The consistency of CT and SPECT/CT (Kappa=0.713) was greater than MRI and SPECT (Kappa=0.449). The sensitivity of CT, MRI, SPECT/CT and MRI-SPECT was 67.1%, 84.5%, 90.8% and 100%, the specificity was 73.3%, 92.3%, 85.6% and 84.6%, and the area under the ROC curve was 0.702, 0.884, 0.882 and 0.923, respectively.@*Conclusions@#SPECT/CT has important impact on clinical management decision for NPC. In the judgement of skull base invasion, the diagnostic value of SPECT/CT is significantly higher than CT and approximately equal to MRI. SPECT/CT should be one of the routine examination methods of nasopharyngeal carcinoma. In addition, in view of its greater diagnostic value, MRI combined with SPECT should be the focus of future imaging studies.
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Objective To study the effect of AG490 on the invasion and metastasis of gallbladder SGC -996 cells regulated by JAK/STAT3,and discuss the related mechanism.Methods Cell viability treated with different concentrations of AG490(50,100,200μmol/L)was detected by MTT method.The ability of invasion and metastasis of gallbladder cells was evaluated by Transwell membrane count.The SGC -996 cell apoptosis was detected by flow cytometry.The protein expression of JAK/STAT3 pathway was detected by Western blot.Results The viability inhi-bition rates of different concentrations of AG490 for SGC -996 cells were (17.49 ±3.41)%,(38.66 ±4.57)%, (79.15 ±6.29)% respectively,and with the increasing of concentration,cell viability decreased obviously.Compared with the control group[(1.39 ±0.21)%],the differences were statistically significant(t =8.162,14.111,21.401, all P <0.01 ).The transfer ability inhibition rate of different concentrations of AG490 for SGC -996 cell were (23.18 ±4.53)%,(51.75 ±6.46)%,(81.32 ±7.13)% respectively,and with the increasing of concentration of AG490,the inhibition rate of invasion and metastasis enhanced.Compared with the control group [(1.46 ± 0.42)%],the differences were statistically significant(t =8.269,13.455,19.366,all P <0.01).The apoptosis rate for SGC -996 cells of different concentrations of AG490 were (13.34 ±4.33)%,(28.16 ±6.23)%,(53.61 ± 8.74)% respectively,and cell apoptosis increased with the increasing of concentration.Compared with control group [(0.97 ±0.52)%],the differences were statistically significant(t =4.913,7.533,10.414,all P <0.01).Different concentrations of AG490 can reduce expression of ZFX,STAT3 and Smad1 protein of JAK/STAT3 pathway of SGC -996 cells,compared with the control group,the differences were statistically significant(tZFX =2.154,3.041,4.185, tSTAT3 =7.348,14.892,17.774 and tSmad1 =3.474,5.241,7.718,all P <0.05).Conclusion AG490 can inhibit the invasion and metastasis of gallbladder SGC -996 cells,and the effect depends on dosage.Its mechanism may be relat-ed to the reduction of cell apoptosis and the protein expression of JAK/STAT3 pathway.
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AIM: To analyze the difference of endonuclease domain containing 1 (ENDOD1) expression be-tween benign prostatic hyperplasia ( BPH ) tissues and prostate cancer ( PCa ) tissues and to investigate the effect of ENDOD1 on the biological function of human prostate cancer cells .METHODS: The BPH samples ( n=20 ) and PCa samples (n=21) were processed and analyzed according to the instruction of immunohistochemical (IHC) staining.The mRNA and protein levels of ENDOD 1 in the normal prostate epithelial cells and prostate cancer cells were evaluated by RT -qPCR and Western blot , respectively .The recombinant plasmids pCMV-N-Flag-ENDOD1 was constructed and was trans-fected into the human prostate cancer cells .The proliferation , apoptosis , migration and invasion abilities of the prostate cancer cells were evaluated by MTT assay , flow cytometry, Transwell migration and Matrigel invasion assays , respectively. RESULTS:The analysis of variance of the immunoreactivity score showed that PCa tissues with high Gleason score dis -played significantly lower ENDOD1expression than that with low Gleason score and BPH (P<0.05).The expression of ENDOD1 at mRNA and protein levels in PC3 cells and DU145 cells was significantly lower than that in the LNCap cells (P<0.05).The proliferation of DU145 transfected with ENDOD1 was inhibited.The flow cytometry indicated that ENDOD1 over-expression in the DU145 cells resulted in a notable increase in G0/G1 phase arrest (P<0.05), but the ap-optotic rates showed no statistical difference .The results of Transwell assay showed that migration and invasion abilities of the cells were also inhibited after transfection with over-expressing ENDOD1 plasmid (P<0.05).CONCLUSION: The expression of ENDOD1 significantly decreased in prostate cancer with high Gleaon score .Meanwhile, the ENDOD1 is spe-cifically down-regulated in androgen independent prostate cancer (AIPC) cell lines.Over-expression of ENDOD1 remark-ably inhibits the proliferation , migration and invasion abilities of AIPC .
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AIM:To investigate the expression of up-regulated gene 11 ( URG11 ) in prostate cancer cell line and the effect of URG11 siNRA on the proliferation and invasion of human prostate cancer LNCaP cells.METHODS:The mRNA and protein levels of URG11 in prostate cancer cell lines and normal prostate epithelial cell line were evaluated by real-time PCR and Western blot.LNCaP cells were transfected with designed siRNA using the liposome method.The prolif-eration, apoptosis, migration and invasion abilities of the LNCaP cells were evaluated by MTS assay, flow cytometry, wound-healing assay and Transwell assay.RESULTS:The expression of URG11 at mRNA and protein levels in the DU145, PC3, LNCaP cell lines was significantly higher than that in RWPE-1 cell line.Compared with the control group, the proliferation of LNCaP cells with URG11 siRNA was stagnant in G1/S phase and induced apoptosis.The proliferation of LNCaP cells at 0 h, 24 h, 48 h and 72 h was inhibited after URG11 expression was down-regulated (P<0.05).Transwell assay showed that migration (P<0.05) and invasion (P<0.05) were also inhibited.CONCLUSION:URG11 is highly expressed in prostate cancer.Silencing of URG11 significantly inhibits the proliferation and invasion and induces apoptosis of LNCaP cells.
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Objective To explore whether hypoxia could promote epithelial-mesenchymal transition (EMT) in various differentiated colorectal cancer cells, and analyse the effect of hypoxia on invasion and migration of colorectal cancer cells. Methods HCT116 (poorly differentiated) and HT-29 (highly differentiated) colorectal adenocarcinoma cells were selected respectively. The morphological changes of two cell lines were observed after 0,10,25,50,100 and 150 mg/L cobalt chloride (CoCl2) treatment for 48 h. The expression of hypoxia-inducible factor-1α(HIF-1α) protein was analysed after 0, 10,25,50,100 and 150 mg/L CoCl2 treatment for 48 h. An optimal concentration of CoCl2 was then selected. Methylthiazolyl tetrazolium (MTT) assay was used to detect the proliferation of two kinds of colorectal cancer cells induced by CoCl 2 at different time points (0, 24, 48, 72 and 96 h), and to select an optimal time. Under the optimal concentration and time conditions, the HCT116 and HT-29 cells were processed by hypoxia (hypoxia group) and normoxia (normoxic group). Transwell invasion assay and Wound healing assay were used to detect cell invasion and migration in two groups. Western blot assay and RT-PCR were used to detect protein and mRNA expression levels of HIF-1α, E-cadherin and Vimentin in two groups. Results Two kinds of cells showed obvious morphological changes after 50 mg/L CoCl2 treatment for 48 h. HIF-1αprotein level first increased and then decreased in two groups of cells with the increased concentration of CoCl 2, and 50 mg/L CoCl2 was the optimal concentration (P<0.05). The cell proliferation showed a tendency to decrease after the increase in both kinds of cells with or without hypoxia for 0-96 h (P<0.05), and 48 h was the optimal time. The transmembrane number and cell migration rate were significantly more in hypoxia group than those of normoxic group (P<0.05). The protein and mRNA levels of HIF-1α and Vimentin were significantly higher in hypoxia group than those of normoxic group in HCT116 and HT-29 cell lines (P<0.05). E-cadherin protein and mRNA levels were significantly lower in hypoxia group than those of normoxic group (P<0.05). Conclusion Hypoxia can promote EMT in different differentiated colorectal cancer cells, and can enhance invasion and migration of two kinds of colorectal cancer cells.
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Objective To explore the expression of urothelial carcinoma-associated 1 ( UCA1 ) in pancreatic cancer cell lines and its influence on the invasion and metastasis of the pancreatic cancer cells .Methods The expression of UCA1 in pancreatic cancer tissues and paired adjacent normal tissues ( 11 cases ) and 5 pancreatic cancer cell lines was analyzed by real-time PCR.The level of UCA1 in BxPC-3 was knocked down by small interfering RNA . The ability of invasion and migration in vitro of transfected BxPC-3 was detected by Transwell invasion assay and wound healing assay .The protein levels of MMP-2 and MMP-9 were measured by Western blot experiment .Results The expression level of UCA1 in pancreatic cancer tissues was higher than that in paired adjacent normal tissues , and UCA1 differentially expressed in 5 pancreatic cancer cell lines .Down-regulation of UCA1 by siRNA suppressed the expression of MMP-2 and MMP-9 in BxPC3, and dramatically impaired the ability of invasion and migration of BxPC-3.Conclusions UCA1 is over-expressed in pancreatic cancer , and down-regulation of UCA1 attenuates the capacity of invasion and metastasis in vitro of BxPC-3 by decreasing MMP-2 and MMP-9.
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[ ABSTRACT] AIM:To compare the expression of SIRT2 in ovarian surface epithelial ( OSE) cell line and serous ovarian carcinoma ( SOC) cell lines, and to investigate the effects of SIRT2 on the cell proliferation, migration and inva-sion.METHODS:The expression levels of SIRT2 in the OSE cell line and the SOC cell lines were determined by Western blot.The SIRT2 siRNAs and overexpression construct were designed and verified.Transient transfection of SIRT2 siRNAs or overexpression construct was performed, and the effect of SIRT2 on the cell proliferation, migration and invasion was e-valuated.RESULTS:SIRT2 levels in the 5 strains of SOC cell lines were significantly lower than that in the OSE cell line.SIRT2 knockdown in HOSEpiC cells significantly enhanced the ability of cell colony formation and accelerated the cell growth rate.On the contrary, overexpression of SIRT2 in HO8910 cells dramatically repressed the number of cell colonies and cell activity.SIRT2 significantly changed the ability of ovarian cell migration.Knockdown of SIRT2 facilitated the cell invasion.CONCLUSION:The expression of SIRT2 in the SOC cells is significantly down-regulated.In the OSE cells, SIRT2 acts as a tumor suppressor and mediates the inhibition of cell proliferation, migration and invasion.
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[ ABSTRACT] AIM:To explore the expression pattern of microRNA-205 ( miR-205) in glioma tissues and its role in the invasion of glioma cells.METHODS:The expression of miR-205 and TBX18 was detected by real-time PCR and immunohistochemical observation, respectively.Transwell assay was used to examine the invasion change of U251 glioma cells after miR-205 overexpression via miR-205 mimics or decrease in miR-205 expression by miR-205 inhibitor.The target of miR-205 was searched by bioinformatics analysis combined with experimental analysis.The protein level of TBX18 was determined by Western blotting after siRNA transfection and Transwell assay was conducted.RESULTS:miR-205 expres-sion was downregulated in 82.6%of detected glioma tissues and TBX18 was significantly overexpressed in glioma tissues compared with normal tissues.miR-205 overexpression remarkably inhibited the invasion potential of U251 glioma cells with a decrease in the invasive cells (P<0.01), while inhibition of miR-205 significantly enhanced the invasion ability of U251 cells.Mechanically, miR-205 directly targeted TBX18 and downregulation of TBX18 also significantly inhibited the invasion potential of U251 cells with a decrease in the invasive cells ( P<0.01 ) .CONCLUSION: miR-205 expression is de-creased in glioma, and miR-205 inhibits glioma cell invasion via targeting TBX18.Our research contributes to the mecha-nisms responsible for glioma invasion and provides theoretical base for developing new therapeutic strategy to treat glioma.