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Objective To investigate the relationship between the expression of CENPF in NSCLC adenocarcinoma (LUAD) and the clinical prognosis of patients and its effect on the metastasis of lung adenocarcinoma cells. Methods The expression of CENPF in LUAD and its relationship with patient prognosis were analyzed by online bioinformatics. The expression of CENPF was verified by LUAD tissue microarray immunohistochemical staining. Kaplan-Meier analysis was performed to analyze the relationship between the expression of CENPF and the prognosis of patients with lung adenocarcinoma. Cox survival hazard ratio was used to analyze the factors affecting the survival of patients. Chi-square analysis was adopted to examine the relationship between CENPF expression and clinicopathological stage and grade of patients. The expression of CENPF in NCI-H2126 cells were knocked out by lentivirus, and then the proliferation, invasion, and migration abilities of the cells were detected. Changes in mRNA expression profiles after CENPF knockout were detected by RNA-seq. Bioinformatics analysis of downstream signaling pathways and the target genes of CENPF was also performed. Western blot was used to verify the target gene. Results CENPF was significantly upregulated in LUAD tumor tissue (P < 0.05) and significantly correlated with pathological stage (P=0.013). The higher expression of CENPF, the worse the prognosis of patients (P=0.01, P=0.027). After the expression was CENPF of knocked out, the cell proliferation, migration, and invasion abilities significantly reduced (P < 0.01). The expression of chemokine pathway genes in cells was enriched significantly (P < 0.001). ACKR3/CXCR7 and CDH2/N-cadherin were significantly downregulated, whereas CDH1/E-cadherin was significantly upregulated. After CENPF was knocked out, ACKR3/CXCR7 and N-cadherin were significantly downregulated, whereas E-cadherin significantly increased. Conclusion The expression of CENPF is negatively correlated with the clinical prognosis of patients with LUAD, and it promotes the occurrence of EMT by regulating the expression levels of N-cadherin and E-cadherin related to EMT through ACKR3/CXCR7.
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OBJECTIVE@#This study aimed to investigate the molecular mechanism of RAB1A in the proliferation, invasion, and metastasis of human tongue squamous cell carcinoma.@*METHODS@#Western blot was used to detect the expression of RAB1A protein in human normal tongue epithelial cells (Hacat) and tongue squamous cell carcinoma Tca8113. The changes in RAB1A after plasmid transfection were also studied. The Tca8113 cells were named SiRAB1A/Tca8113 after RAB1A plasmid transfection. The expression of the epithelial-mesenchymal transition (EMT)-related markers of SiRAB1A/Tca8113 cells was also detected. CCK-8 assay was used to detect the proliferation of SiRAB1A/Tca8113 cells. Transwell and wound healing assays were used to detect the invasive and metastatic abilities of SiRAB1A/Tca8113 cells, respectively.@*RESULTS@#Western blot results showed that the expression of RAB1A in tongue squamous cell carcinoma cells was significantly higher than that in Hacat. RAB1A decreased significantly after SiRAB1A plasmid transfection. CCK-8 proliferation assay showed that the proliferation of SiRAB1A/Tca8113 cells also decreased significantly. Transwell and wound healing assays demonstrated that the invasive and metastatic abilities of SiRAB1A/Tca8113 cells decreased significantly, respectively. In addition, Western blot results demonstrated that RAB1A deletion significantly increased the expression of E-cadherin and inhibited the expression of Vimentin.@*CONCLUSIONS@#RAB1A could promote the proliferation, invasion, and metastasis of tongue squamous cell carcinoma cells.
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Humans , Carcinoma, Squamous Cell , Cell Line, Tumor , Cell Movement , Cell Proliferation , Epithelial-Mesenchymal Transition , Head and Neck Neoplasms , Tongue NeoplasmsABSTRACT
Objective To study the expression of Stat5,E-cadherin and Vimentin in thyroid carcinoma and the correlation among them. Method The expression of Stat5,E-cadherin and Vimentin in 149 cases of thyroid specimens(including 23 cases of nodular goiter and 126 cases of thyroid carcinoma)was detected by immunohisto-chemistry SP method and the correlation analysis was conducted with the main clinical pathological parameters. Results The positive rate of Stat5 and Vimentin in thyroid carcinoma was 85.71% and 77.78% respectively, which was significantly higher than that in nodular goiter(26.09%and 8.70%)(P 0.05),but obviously with lymph node metastasis and pathological type(P<0.05). Stat5 was negatively correlated with the expression of E-cadherin (r=-0.335,P=0.000)but positively with the expression of Vimentin(r=0.218,P<0.05). Conclusion The high expression of Stat5 and EMT in thyroid carcinoma tissues indicate that Stat5 may be involved in the EMT process of thyroid carcinoma ,thus it can promote the invasion and metastasis of thyroid carcinoma.
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Background and purpose:Human epidermal growth factor receptor-2 (HER-2), a member of epidermal growth factor receptor family, initiates a diverse set of signaling pathways that ultimately affect such fun-damental processes as cell proliferation, cell motility and cell apoptosis. It is reported that HER-2 was associated with epithelial-mesenchymal transition (EMT) process. However, the mechanism needs further investigation. The purpose of this study was to investigate the mechanism of HER-2 on regulating EMT process.Methods:Transwell assay was used to determine the motility of breast cancer cells; Real-time lfuorescence quantitative polymerase chain reaction (RT-FQ-PCR) was employed to determine the expression of genes of interest, and reactive oxygen species production was measured by reactive oxygen species detection kit.Results:HER-2 overexpression in breast cancer cells could promote cell migration and invasion. Mechanistic study showed that HER-2 overexpression could upregulate ZEB1 expression. ZEB1 silencing by siRNA reduced cell motility of HER-2-overexpressing breast cancer cells. Furthermore, reactive oxygen species produced in HER-2-overexpressing breast cancer cells were less than those produced in corresponding control cells.Conclusion:Our study demonstrated that HER-2 overexpression endowed breast cancer cells with EMT related properties by upregulating ZEB1 expression. ZEB1 could be a candidate target for further study of the relation-ship between HER-2 and EMT.
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Objective To explore the role of JNK signaling pathway in epithelial mesenchymal transition (EMT)process of human alveolar epithelial cells A549 induced by TGF‐β1 in vitro.Methods Human alveolar epithelial cells (A549)cultured in vitro were divided randomly into three groups :normal group ,TGF‐β1 group ( treated by TGF‐β1 with 10 ng/mL)and inhibitor group (treated by 10 ng/mL TGF‐β1 and 20 μmol/L Sp600125).Morphological observation on the cells was performed under light microscope after culturing in 3% serum medium for 72 h. The expression of E‐cadherin (E‐cad ,a epithelial cell marker) ,α‐smooth muscle actin (α‐SMA ,a mesenchymal cell marker)and Collagen fibers Ⅰ(ColⅠ ,a mesenchymal cell marker)were tested by RT‐PCR.The level of JNK phosphorylation (p‐JNK)was detected through Western blot.All experiments were repeated three times at least.Results The normal human alveolar epithelial cells (A549)cultured invitro were arranged closely like peb‐bles.E‐cad expressed at a certain level ,while the expression ofα‐SMA ,ColⅠand p‐JNK was weakly detected.In TGF‐β1 group , the cells were spindle‐shaped ,the expression of E‐cad was reduced ,while the expression ofα‐SMA ,ColⅠand p‐JNK was signifi‐cantly increased 72 h after treatment of TGF‐β1. However ,compared with TGF‐β1 group ,spindle‐shaped cells in the inhibitor group were recovered after 72 h being treated by TGF‐β1 and Sp600125 ,the expression of E‐cad was increased and the expres‐sion levels of α‐SMA ,ColⅠand p‐JNK were significantly decreased 72 h after treatment with TGF‐β1 and Sp600125 in inhibitor group. As compared with normal group ,the shape of the cells in inhibitor group was prolate ,and the expression of E‐cad ,α‐SMA ,ColⅠand p‐JNK was not significantly different.Conclusion JNK signaling pathway is related to the process of EMT of human alveolar epithelial cells induced by TGF‐β1. Sp600125 ,a special inhibitor of JNK ,could validly restrain the process.