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Objective: To observe the expression of valosin-containing protein CVCP) in the epithelial ovarian cancer (EOC) tissue and its relationships with the clinicopathological features of the EOC patients∗ and to provide the basis for the molecular treatment of EOC. Methods: The expressions of VCP in 94 EOC tissue samples, 13 ovarian borderline tumor tissue samples, 36 ovarian benign tumor tissue samples and 8 normal ovarian tissue samples were measured by immunohistochemical method. The relationships between the VCP expression and the clinicopathological parameters (age, FIGO stage, pathological type, histological grade, lymph node metastasis or not, ascites or not, preoperative CA125 level) of the EOC patients were analyzed. The expressions of VCP in human ovarian epithelial HOSEPIC cells and human EOC SKOV-3 cells were detected by immunofluorescence technique and Western blotting method. The SKOV-3 cells were divided into control group (without CB-5083) and treatment group (with CB-5083)∗ the migration rates of the SKOV-3 cells in two groups were analyzed by scratch experiment, and the clone formation rates of the SKOV-3 cells in two groups were analyzed by plate clone formation experiment. The expressions of VCP in the cells in two groups were analyzed by immunofluorescence technique. The expression levels of VCP, NF-kB/P65» IieBa∗ and p-Iiefta proteins in the SKOV-3 cells in two groups were detected by Western blotting method. Results: The positive expression rates of VCP in EOC, borderline ovarian tumor, benign ovarian tumor and normal ovarian tissues had significant difference ( P0. 05). The VCP expression level in the SKOV-3 cells was higher than that in HOSEpiC cells ( P<0. 05). The migration rate of the SKOV-3 cells in treatment group was lower than that in control group ( P<0.05), the cloning formation rate of the SKOV-3 cells in treatment group was lower than that in control group ( P<0. 05). The expression levels of VCP and NF-kB/P65 proteins of the SKOV-3 cells in treatment group were lower than those in control group (P'<0. 05) ∗ and the expression level of p-IieBa protein was higher than that in control group ( P< 0. 05). Conclusion: VCP is highly expressed in the EOC tissue and cells∗ and high-expression VCP can promot the tumor migration and proliferation.
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Objective To prokaryotically express the valosin-containing protein(VCP)of Schistosoma japonicum,and ana-lyze its VCP mRNA expressions in the cercaria,schistosomulum,adult worm(female and male worms)and egg. Methods RNA of S. japonicum eggs were extracted,and reversely transcribed into cDNA. The VCP gene of S. japonicum was amplified by using polymerase chain reaction(PCR),and subcloned into the prokaryotically expressed vector pET15b. The recombined plasmid was transformed into BL21 cells,and the expression of the target gene was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG). The recombinant protein was yielded through the purification of inclusion body,and identified by using sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The RNA(s)of cercaria,schistosomulum,female adult worm,male adult worm,and egg of S. japonicum were extracted,digested with DNase,purified,and reversely transcribed into cDNA. The mRNA expressions of the VCP gene in various developmental stages of S. japonicum were determined by using fluorescence-based quantitative real-time PCR. Results The VCP gene of S. japonicum was yielded by PCR amplification,and the recombinant pro-tein was obtained through recombinant plasmid expression and purification of inclusion body. The highest VCP mRNA expression in S. japonicum cercaria was detected by the fluorescence-based quantitative real-time PCR,while low expressions were found in the schistosomulum,egg,female and male adult worms. Conclusion The recombinant protein encoded by the VCP gene of S. ja-ponicum is successfully obtained,and the VCP mRNA expression is determined in various developmental stages of S. japonicum.
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Objective High expression of the valosin-containing protein ( VCP) gene can enhance the metastasis of osteosar-coma via the AKT/PI3K/NF-KappaB/MMP-9 signaling pathway, but the molecular mechanisms underlying the up-regulation of VCP in osteosarcoma cells remains unknown .This study aimed to determine whether miRNA-129-5p can regulate the VCP expression and its targets in human osteosarcoma cells . Methods The microRNA target-predicting software TargetScanhuman 6.2 ( http://www.tar-getscan.org/) was used to predict the possible targets of miRNA-129-5p on the VCP gene.Then, two recombinant gene report vectors containing the wild VCP gene 3′UTR ( psi-VCP vector ) and mutant VCP gene 3′UTR ( psi-VCPmut vector ) were constructed , se-quenced, and identified.The human osteosarcoma U2-OS cells were co-transfected with miRNA-129-5p mimic and psi-VCP vector or psi-VCPmut vector, respectively.A non-specificity mimic transfection served as negative control , and the luciferase activity was detec-ted in each group. Results The software prediction showed only one conserved function site of miRNA-129-5p on the VCP gene 3′UTR163-169 bp.Luciferase activity was significantly lower in the psi-VCP vector +miRNA-129-5p transfection group (15.529 ± 1.902) than in the VCP control group (21.781 ±0.854), VCP mutation experimental group (19.978 ±1.377), and VCP mutation control group (21.952 ±1.516) (P<0.05), with no remarkable difference between the VCP mutation control and VCP control groups (P=0.276). Conclusion miRNA-129-5p can probably regulate the targets of the VCP gene in human osteosarcoma U 2-OS cells.
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BACKGROUND: Inclusion-body myopathy with Paget's disease of the bone and frontotemporal dementia (IBMPFD) is a rare, late-onset autosomal disorder arising from missense mutations in a gene coding for valosin-containing protein. CASE REPORT: We report the case of a man carrying the previously described p.Arg159His mutation, who had an unusual axonal sensorimotor neuropathy as the first clinical manifestation of IBMPFD, and for whom diagnosis only became clear 8 years later when the patient developed frontotemporal dementia. CONCLUSIONS: Peripheral neuropathy is a rare manifestation of IBMPFD. This underdiagnosed disorder should be considered when a patient develops dementia or has signs of Paget's disease.
Subject(s)
Humans , Axons , Central Nervous System , Clinical Coding , Dementia , Diagnosis , Frontotemporal Dementia , Genes, vif , Muscular Diseases , Mutation, Missense , Peripheral Nervous System DiseasesABSTRACT
Background and Aim: The strategies of targeting valosin-containing protein (VCP) may have therapeutic potential for treating cancer metastasis. In this study, we aim to investigate the correlation of VCP protein expression in osteosarcoma (OS) tissues with pulmonary metastasis and its possible molecular mechanism. Materials and Methods: Expression of VCP in 60 OS specimens was detected by immunohistochemistry (IHC) and the relationship with metastasis was analyzed. An artifi cial micro ribonucleic acid, targeting VCP, was performed to silence the expression of VCP in U2-OS cells. Cell mobility was detected by wound healing and Transwell assays. Western blot and real-time polymerase chain reaction were performed to investigate the expression of VCP in U2-OS cells. Furthermore, the protein of pAKT (phosphorylated serine/threonine protein kinase) and nuclear factor of kappa B protein65 were measured by western blot to evaluate the effect of silencing VCP on AKT/nuclear factor of kappa B (NF-B) signaling pathway. Results: The results showed that the expression level of VCP protein in cases with pulmonary metastases was signifi cantly higher than that in those without metastasis (P = 0.004). The invasion and migration of U2-OS cells were suppressed by silencing VCP. Furthermore, silencing VCP could down-regulate the phosphorylation of AKT and nuclear transfer of NF-B. Conclusions: Our fi ndings suggested that inhibition of VCP could suppress OS cells invasion and migration through down-regulating AKT/NF-B signaling pathway.