ABSTRACT
OBJECTIVE: To verify whether micro ribonucleic acid 34-a can exert its negative effects in human neuroblastoma cells. METHODS: The study was conducted at The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China during 15 months (from March 2015 to about June 2016). Quantitative reverse transcription polymerase chain reaction was used to find the differences of micro ribonucleic acid 34-a between metastatic neuroblastoma and primary tumours. We transfected micro ribonucleic acid 34-a mimics and antisense oligonucleotides into neuroblastoma cell line to explore its function in vitro through the variations. Additionally, fluorescent reporter assay was used to clear the targeting site of micro ribonucleic acid 34-a and CD44. Furthermore, protein levels of CD44, the putative target gene of micro ribonucleic acid 34-a, was assessed after transfection by Western blot. RESULTS: Compared to the primary neuroblastoma tumours, micro ribonucleic acid 34-a was lower in metastatic neuroblastoma using quantitative reverse transcription polymerase chain reaction (p<0.05). Transfection of micro ribonucleic acid 34-a mimics and antisense oligonucleotides into a neuroblastoma cell line significantly affected cellular activity, migration and invasion (p<0.05_. Fluorescent reporter assays proved that CD44 acts as the target spot of micro ribonucleic acid 34-a for repression in post-transcription level. Micro ribonucleic acid 34-a inhibited the expression of CD44, and increased concentration of micro ribonucleic acid 34-a mimics resulted in a greater decrease in the expression of CD44. CONCLUSIONS: Micro ribonucleic acid 34-a might suppress the progression of neuroblastoma through inhibiting the expression of the potential target gene CD44.
Subject(s)
Hyaluronan Receptors/metabolism , MicroRNAs/metabolism , Neuroblastoma/metabolism , Neuroblastoma/pathology , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Humans , Hyaluronan Receptors/analysis , Hyaluronan Receptors/genetics , MicroRNAs/pharmacologyABSTRACT
In this study, we found that the expression of miR-15a was positively correlated with neuroblastoma (NB) clinical pathological stage and was negatively correlated with reversion-inducing cysteine-rich protein with Kazal motifs (RECK) expression. Using the enhanced green fluorescent protein (EGFP) reporter construct carrying the 3'-UTR of RECK, we identified RECK as a direct target of miR-15a. Suppression of miR-15a significantly decreased the migration ability of GI-LA-N and SK-N-SH cell lines, whereas overexpression of miR-15a increased the migration ability; these effects could be partly reversed by RECK inhibition or ectopic expression. Moreover, inhibition of miR-15a significantly increased secreted matrix metalloproteinase-9 expression in culture medium through regulating the expression of RECK. These findings provide new insights into the characteristics of the miR-15a-RECK-matrix metalloproteinase-9 axis in NB progression, especially in NB migration and invasion.
Subject(s)
GPI-Linked Proteins/genetics , Matrix Metalloproteinase 9/genetics , MicroRNAs/genetics , Neuroblastoma/genetics , 3' Untranslated Regions/genetics , Blotting, Western , Cell Line, Tumor , Cell Movement/genetics , GPI-Linked Proteins/metabolism , Gene Expression Regulation, Neoplastic , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , HEK293 Cells , HeLa Cells , Humans , Matrix Metalloproteinase 9/metabolism , Models, Genetic , Neoplasm Invasiveness , Neoplasm Metastasis , Neoplasm Staging , Neuroblastoma/metabolism , Neuroblastoma/pathology , RNA Interference , Reverse Transcriptase Polymerase Chain Reaction , Spectrometry, FluorescenceABSTRACT
The purpose of this study was to investigate and discuss imaging methods and management strategies for congenital choledochal cyst with co-existing intrahepatic dilation and aberrant bile duct as well as other complicated biliary anomalies. In this study we reviewed and analyzed 72 patients with congenital choledochal cyst, ranging in age from 15 days to 12 years old and who were seen at our hospital during the past 12 years, from January 1993 to October 2005. The image manifestation and clinical significance of patients with co- xisting intrahepatic biliary dilation and aberrant bile duct were carefully examined during operation via MRCP, cholangiography and choledochoscope. Twenty-two cases (30.1%) presented with intrahepatic bile duct dilation and 12 of these were of the cystic type. That is, the orifice of the dilated intrahepatic tract that converged into the common hepatic duct showed membrane or septum-like stenosis. In 10 cases the dilation tapered off from the porta hepatis to the initiating terminals of the intra-hepatic bile ducts and was not accompanied by stenosis. An aberrant bile duct was observed in 2 of the cases. In 3 cases, the right and left hepatic ducts converged at the choledochal cyst. In conclusion, the imaging methods for intrahepatic bile duct dilation possess important clinical significance. Further, for hepatojejunostomy with radical excision of a choledochal cyst, additional operative procedures for intrahepatic stenosis, possible bile duct malformation and pancreaticobiliary common duct calculi can potentially reduce postoperative complications.
Subject(s)
Male , Infant, Newborn , Infant , Humans , Female , Child, Preschool , Child , Tomography, X-Ray Computed , Postoperative Complications/diagnostic imaging , Liver Diseases/complications , Choledochal Cyst/complications , Cholangiography , Bile Ducts/abnormalitiesABSTRACT
The purpose of this study was to investigate and discuss imaging methods and management strategies for congenital choledochal cyst with co-existing intrahepatic dilation and aberrant bile duct as well as other complicated biliary anomalies. In this study we reviewed and analyzed 72 patients with congenital choledochal cyst, ranging in age from 15 days to 12 years old and who were seen at our hospital during the past 12 years, from January 1993 to October 2005. The image manifestation and clinical significance of patients with co- xisting intrahepatic biliary dilation and aberrant bile duct were carefully examined during operation via MRCP, cholangiography and choledochoscope. Twenty-two cases (30.1%) presented with intrahepatic bile duct dilation and 12 of these were of the cystic type. That is, the orifice of the dilated intrahepatic tract that converged into the common hepatic duct showed membrane or septum-like stenosis. In 10 cases the dilation tapered off from the porta hepatis to the initiating terminals of the intra-hepatic bile ducts and was not accompanied by stenosis. An aberrant bile duct was observed in 2 of the cases. In 3 cases, the right and left hepatic ducts converged at the choledochal cyst. In conclusion, the imaging methods for intrahepatic bile duct dilation possess important clinical significance. Further, for hepatojejunostomy with radical excision of a choledochal cyst, additional operative procedures for intrahepatic stenosis, possible bile duct malformation and pancreaticobiliary common duct calculi can potentially reduce postoperative complications.
