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Objective To analyze the influence of cyclic RNA homologous domain interacting protein kinase 3(circ_HIPK3)on function and morphology of myloid β-protein(Aβ)induced hippocampal neurons by targeting miR-381-3p/zinc finger protein 217(ZNF217)axis.Methods Hippocampal neurons of neonatal rats were prepared and divided into the control group,the Aβ group,the si NC1 group,the si HIPK3 group,the si HIPK3+inhibitor NC group,the si HIPK3+miR-381-3p inhibitor group,the si HIPK3+miR-381-3p inhibitor+si NC2 group and the si HIPK3+miR-381-3p inhibitor+si ZNF217 group.Except the control group,all the other groups were modeled by 40 μmol/L Aβ1～42.qRT-PCR was used to determine the circ of hippocampal neurons circ_HIPK3,miR-381-3p and ZNF217 mRNA levels.Cell morphology was observed by transmission electron microscope,and the survival rate of hippocampal neurons was measured by CCK-8 method.Hochesst 33342 method was used to measure apoptosis of hippocampal neurons.The intracellular Ca2+ fluorescence intensity of hippocampal neurons was detected by flow cytometry.The expression levels of P-Tau,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),Caspase-3 and ZNF217 proteins in hippocampal neurons were measured by Western blot assay.Double luciferase reporter genes were used to analyze the targeting relationship between miR-381-3p and circ_HIPK3,ZNF217.Results In the control group,the structure of hippocampal neurons was normal,the morphology of nucleus was normal,and there were no pathological changes in mitochondria and endoplasmic reticulum.In the Aβ group,hippocampal neurons showed degenerative changes,abnormal nuclear morphology,membrane invagination,a large number of mitochondria swelling and a large number of lipid droplets vacuoles in cytoplasm.Compared with the Aβ group,the hippocampal neuronal structure was partially restored in the si HIPK3 group.Compared with the si HIPK3 group,the hippocampal neuronal structure was severely damaged in the si HIPK3+miR-381-3p inhibitor group.Compared with the si HIPK3+miR-381-3p inhibitor group,the damage of hippocampal neurons in the si HIPK3+miR-381-3p inhibitor+si ZNF217 group was reduced.Compared with the control group,the circ_HIPK3,ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax,Caspase-3 protein expression of hippocampal neurons were increased in the Aβ group,and the miR-381-3p level,survival rate and Bcl-2 protein expression decreased(P＜0.05).Compared with the Aβ group,the circ_HIPK3,ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax and Caspase-3 protein expression of hippocampal neurons were decreased in the si HIPK3 group,and miR-381-3p level,survival rate and Bcl-2 protein expression increased(P＜0.05).Compared with the si HIPK3 group,the circ_HIPK3,ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax and Caspase-3 protein expression of hippocampal neurons in the si HIPK3+miR-381-3p inhibitor group were increased,and the miR-381-3p level,survival rate and Bcl-2 protein expression decreased(P＜0.05).Compared with the si HIPK3+miR-381-3p inhibitor group,the ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax and Caspase-3 protein expression of hippocampal neurons in the si HIPK3+miR-381-3p inhibitor+si ZNF217 group were decreased,and the survival rate and Bcl-2 protein expression increased(P＜0.05).miR-381-3p targeted and combined with HIPK3 and ZNF217.Conclusion circ_HIPK3 silencing may ameliorate Aβ-induced damage of hippocampal neuronal structure and function by regulating miR-381-3p/ZNF217 axis.
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Objective To determine the expression of ZNF217 protein in bladder cancer tissues and to analyze its clinicopathological characteristics, so as to clarify its clinical significance. Methods The clinicopathological data of 123 patients with bladder cancer who underwent bladder cancer resection were retrospectively analyzed. The expression of ZNF217 protein in bladder cancer tissues and corresponding adjacent tissues was detected by immunohistochemistry, and its correlation with clinical characteristics of patients was analyzed. Results ZNF217 protein was highly expressed in tumor tissues, and the ratio of high expression of tumor tissues was 67.4％, but it was lowly expressed in adjacent tissues. By comparing with clinicopathological features, the expression level of ZNF217 was significantly correlated with tumor T stage and tumor recurrence (all P<0.05), but not correlated with age , gender , tumor differentiation and lymph node metastasis (all P>0.05). The survival analysis showed that patients with high expression of ZNF217 in bladder cancer had significantly shorter overall survival and progression-free survival than those with low ZNF217 expression ( all P<0 . 05 ) . Conclusions ZNF217 is highly expressed in bladder cancer tissues, and its expression level is associated with tumor stage, tumor recurrence and poor prognosis, suggesting that it can be used as a potential therapeutic target for bladder cancer.
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Objective To investigate the expression and clinical significance of zinc finger protein 217 (ZNF217)in human pancreatic cancer.Methods 43 cases with pancreatic cancer undergoing surgery in the PLA 117 Hospital and People's Hospital of Ruian City from Apr .2011 to May.2014 were enrolled in the study . The pancreatic cancer and the corresponding tumor-adjacent tissues were collected .Real-time quantitative PCR (qRT-PCR)was applied to detect ZNF217 mRNA expression in pancreatic cancer (n=43)and the corresponding tumor-adjacent normal tissues.The protein expression of ZNF217 was measured by immunohistochemistry(IHC). The relationship between the expression of ZNF 217 and clinical features was analyzed by pearson chi-square test . Results The expression level of ZNF217 mRNA and protein was significantly higher in pancreatic cancer tissues than in adjacent normal tissues(P<0.05).The high expression of ZNF217 protein was positively correlated with perineural invasion, tumor size, lymphatic metastasis and advanced TNM stage (P<0.05).Conclusions The expression of ZNF217 is significantly higher in pancreatic cancer tissues than in tumor-adjacent normal tissues , and the upregulation of ZNF 217 is associated with clinicopathological features of tumor malignance .ZNF217 may become a new marker and effective therapeutic target in early diagnosis of pancreatic cancer .
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Objective To study the expression of ZNF217 and EF1α gene in the pathological scars and to investigate role and probable mechanism in the pathogenesis of abnormal scar.Methods Quantitative real-time PCR and Western blot were performed to detect the expression and distribution of mRNA and protein of ZNF217 and EF1α in hypertrophic scar (10 cases),keloid (10 cases),normal scar (10 cases),and normal skin (10 cases),and statistics was used to analyze the data.Results The expression of ZNF217 mRNA and protein in the normal skin,normal scar,hypertrophic scar and keloid were 1.46±0.397,1.45±0.265,4.49±0.999,5.47±0.808; 0.276±0.0211,0.299±0.0150,0.743t0.0509 and 0.747±0.0377,respectively.The expression of EF1α mRNA and prorein in the normal skin,normal scar,hypertrophic scar,and keloid were 1.47±0.469,1.47±0.218,5.10±1.68,5.74±1.92; 0.505±0.0371,0.518±0.0153,0.780±0.0369 and 0.792±0.0290,respectively.The positive rate of mRNA and protein of ZNF217 and EF1α was not statistically different between the hypertrophic scar and keloid (P＞0.05),while they were all remarkably significant in comparison between normal scar and abnormal scar (P＜0.01).In pathological scar mRNA and protein of ZNF217 and EF1α showed a strong positive correlation.Conclusions The expression of ZNF217 and EF1α is increased in pathological scar.Therefore,ZNF217 and EF1α overexpression may play an important role in the proliferation of fibroblasts and in the pathogenesis of pathological scar.
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Objective To explore the mRNA and protein expressions of zinc-finger protein 217(ZNF217)in cisplatin(DDP)-sensitive and DDP-resistant human ovarian cancer cell lines.Methods Six strains of ovarian cancer cells,including 3 DDP-sensitive strains(A2780,SKOV-3 and COC1)and 3 DDP-resistant strains(A2780-DDP-R,SKOV-3-DDP-R and COC1-DDP-R)were selected.The relative luminescence unit(RLU)of cells was determined with ATP assay,the IC50 value and resistance index(RI)of DDP-resistant cell lines were calculated.Intracellular localization of ZNF217 protein in the 6 ovarian tumor cell lines was detected by immunofluorescent cytochemistry.The expressions of ZNF217 mRNA and protein were determined by RT-PCR and Western blotting,respectively.Results The IC50 values of DDP to A2780 and A2780-DDP-R were 18.1?2.3mg/L and 47.9?3.8mg/L(P