ABSTRACT
BACKGROUND@#In our previous paper, we demonstrated that Connexin 43 (CX43) was highly expressed in bladder cancer (BC) tissues. But the molecular mechanism about microRNAs (miRNAs) regulation upstream of CX43 in BC has not been well elucidated and remains to be further studied. MicroRNA-139-5p (miR-139-5p) is a tumor suppressor in progression of multifarious cancers including BC. Nevertheless, the underlying mechanisms of CX43/miR-139-5p in tumorigenesis of BC are still not well illustrated. The specific objective of our study was to inquiry the effect of CX43/miR-139-5p on BC progression and its underlying mechanism.@*METHODS@#The bioinformatics analysis softwares were applied to predict the miRNAs in the upstream of CX43. First, the expression levels of miR-139-5p in BC tissues (tumor) and paracancer tissues (normal) were investigated using the data from The Cancer Genome Atlas database. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression level of miR-139-5p in three human BC cell lines 5637, T24, ECV-304 and a human bladder epithelial immortalized cell line SV-HUC-1 (normal control). Then si-CX43, si-control, miR-139-5p mimic, and its negative control (NC) were transfected into BC cell line ECV-304. The relationship of miR-139-5p and CX43 was analyzed by dual-luciferase reporter assay. The qRT-PCR and Western blotting were used to test the mRNA and protein expression level of CX43. The proliferation of ECV-304 and T24 cells were examined by cell counting kit-8. The migration and invasion of ECV-304 cells were tested by transwell assay. To determine whether miR-139-5p would affect cell proliferation, migration and invasion by targeting CX43, we executed the rescue assay. The comparison between two groups was analyzed by Student's t test, and comparisons among multiple samples were performed by one-way analysis of variance and a Bonferroni post hoc test.@*RESULTS@#The expression of miR-139-5p was remarkably down-regulated in BC tissues (tumor vs. normal, 2.286 ± 0.017 vs. 3.211 ± 0.034, t = 11.540, P < 0.0001) and cell lines (P < 0.01 in all BC cell lines). Besides, we also indicated that over-expression of miR-139-5p reduced the proliferation of ECV-304 (P = 0.001) and T24 cells (P = 0.005). Moreover, miR-139-5p over-expression weakened the invasion (P = 0.001) and migration (P = 0.001) of ECV-304 cells. Furthermore, the relative luciferase activity of CX43-wild type construct was distinctly lessened by up-regulation of miR-139-5p (miR-139-5p mimic NC vs. miR-139-5p mimic, 0.916 ± 0.063 vs. 0.356 ± 0.048, t = 7.085, P = 0.002), nevertheless the activity of CX43-mutant type construct was untouched (miR-139-5p mimic NC vs. miR-139-5p mimic, 0.918 ± 0.057 vs. 0.878 ± 0.039, t = 0.577, P = 0.595). Finally, the rescue assay revealed that CX43 deletion enhanced the depressor effect of miR-139-5p on ECV-304 cell proliferation (P < 0.01), invasion (P = 0.028), and migration (P = 0.014).@*CONCLUSION@#MiR-139-5p, as a tumor-suppressor, repressed cell proliferation, invasion, and migration in BC, which might be achieved by regulating CX43.
ABSTRACT
Background@#In our previous paper, we demonstrated that Connexin 43 (CX43) was highly expressed in bladder cancer (BC) tissues. But the molecular mechanism about microRNAs (miRNAs) regulation upstream of CX43 in BC has not been well elucidated and remains to be further studied. MicroRNA-139-5p (miR-139-5p) is a tumor suppressor in progression of multifarious cancers including BC. Nevertheless, the underlying mechanisms of CX43/miR-139-5p in tumorigenesis of BC are still not well illustrated. The specific objective of our study was to inquiry the effect of CX43/miR-139-5p on BC progression and its underlying mechanism.@*Methods@#The bioinformatics analysis softwares were applied to predict the miRNAs in the upstream of CX43. First, the expression levels of miR-139-5p in BC tissues (tumor) and paracancer tissues (normal) were investigated using the data from The Cancer Genome Atlas database. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression level of miR-139-5p in three human BC cell lines 5637, T24, ECV-304 and a human bladder epithelial immortalized cell line SV-HUC-1 (normal control). Then si-CX43, si-control, miR-139-5p mimic, and its negative control (NC) were transfected into BC cell line ECV-304. The relationship of miR-139-5p and CX43 was analyzed by dual-luciferase reporter assay. The qRT-PCR and Western blotting were used to test the mRNA and protein expression level of CX43. The proliferation of ECV-304 and T24 cells were examined by cell counting kit-8. The migration and invasion of ECV-304 cells were tested by transwell assay. To determine whether miR-139-5p would affect cell proliferation, migration and invasion by targeting CX43, we executed the rescue assay. The comparison between two groups was analyzed by Student’s t test, and comparisons among multiple samples were performed by oneway analysis of variance and a Bonferroni post hoc test.@*Results@#The expression of miR-139-5p was remarkably down-regulated in BC tissues (tumor vs. normal, 2.286 ± 0.017 vs. 3.211 ± 0.034, t= 11.540, P < 0.0001) and cell lines (P < 0.01 in all BC cell lines). Besides, we also indicated that over-expression of miR-139-5p reduced the proliferation of ECV-304 (P = 0.001) and T24 cells (P = 0.005). Moreover, miR-139-5p over-expression weakened the invasion (P = 0.001) and migration (P = 0.001) of ECV-304 cells. Furthermore, the relative luciferase activity of CX43-wild type construct was distinctly lessened by up-regulation of miR-139-5p (miR-139-5p mimic NC vs. miR-139-5p mimic, 0.916 ± 0.063 vs. 0.356 ± 0.048, t = 7.085, P = 0.002), nevertheless the activity of CX43-mutant type construct was untouched (miR-139-5p mimic NC vs. miR-139-5p mimic, 0.918 ± 0.057 vs. 0.878 ± 0.039, t= 0.577, P = 0.595). Finally, the rescue assay revealed that CX43 deletion enhanced the depressor effect of miR-139-5p on ECV-304 cell proliferation (P < 0.01), invasion (P = 0.028), and migration (P = 0.014).@*Conclusion@#MiR-139-5p, as a tumor-suppressor, repressed cell proliferation, invasion, and migration in BC, which might be achieved by regulating CX43.
ABSTRACT
Objective:To study the liquefaction in the cystic cavity of cerebral infarction and the blood supply in the wall to explore whether it is suitable for neural stem cell transplantation. Methods:A total of 20 male Sprague-Dawley rats were established middle cerebral artery occlusion (MCAO) model. They were observed the formation of cystic cavity three, ten and 21 days after modeling with MRI, and the coordinates of cystic cavity and lateral ventricle were recorded. Intracystic fluid and cerebrospinal fluid were extracted from the rats with cavities for mass spectrometry. Frozen sections of the brains were stained with HE and immunofluorescence agglutinin and observed. Results:The cystic cavity in the infarction area became stable 21 days after modeling, which was composed of the brain tissue and the pia matter. There was no cellular structure in the center of the cystic cavity. The wall of the cystic cavity was partly composed of the pia matter. There were a large number of cells at the junction between the center of the cystic cavity and the meninges. Blood vessels distributed around the cystic cavity, similar with those in unaffected side. A total of 36 different molecules were screened out, in which 31 increased and five decreased. For the top ten molecules, most of them positively benefited stem cell transplantation, including anti-inflammatory, anti-tumor and promoting the survival of nerve cells. However, they did not liked to promote neural stem cells differentiating into neurons, but glial cells. Conclusion:As a relatively closed fluid space, the cystic cavity formed in the chronic cerebral infarction can provide basic conditions for neural stem cell transplantation. However, relevant molecular components in the microenvironment mostly promote the differentiation of neural stem cells into glial cells.
ABSTRACT
Objective To investigate the variation of renal pelvic pressure during percutaneous nephrolithotomy (PCNL) via standard nephrostomy tract and explore its influence on renal function. Methods 156 patients with renal calculi were selected for PCNL in standard-tract. The patients were divided into normal, mild hydronephrosis, moderate hydronephrosis groups according to the image by color Doppler ultrasonograph. A transurethral 6F ureteral catheter was inserted into renal pelvis and connected to the pressure monitering system before PCNL. During the operations, all the nephrostomy tracts were dilated to F24 size after successful puncture. Energy used was pneumatic and ultrasound lithotripsy. Renal function of the patients was evaluated with glomerular filtration rate (GFR) determined by 99mTc-DTPA dynamic renal imaging before and one week after PCNL. Data were analyzed by SPSS 19.0 software. Results The stone clearance rate was 75.0％ in one-session procedure. Severe complications did not occur during the operation, such as hemorrhage needing nephrectomy and abdominal organ injury or pneumothorax. There were no statistically significant differences between normal and mild hydronephrosis groups for the variation of renal pelvic pressure during preoperative versus intraoperative PCNL (P > 0.05). The renal pelvic pressure was significantly higher during operation than those of preoperation in moderate hydronephrosis group (P < 0.05), and it was greater than those of normal and mild hydronephrosis groups during operation (P < 0.05). Renal pelvic pressure generally remained lower than a level to 30.00 mmHg. There were no significant differences of preoperative and postoperative glomerular filtration rate in all the groups (P > 0.05). Conclusions There were no significant differences on the renal pelvic pressure in normal group and mild hydronephrosis group during operation via standard nephrostomy tract. It should be careful to maintain the lower intrapelvic pressure in order to avoid reflux and infection in moderate hydronephrosis group. Percutaneous nephrolithotomy via standard- tract does not cause significant effects on glomerular filtration rate during the perioperative period of PCNL .