ABSTRACT
Fibroblast growth factor receptor 1 (FGFR1) has been reported in gastric cancer to be a prognostic factor. However, miR-497-targeted FGFR1 has not been explored in the carcinogenesis of gastric cancer. The present study intended to revalidate the prognostic significance of FGFR1 in patients with gastric cancer, and the mechanism of miR-497-regulated FGFR1 was investigated in gastric cancer cell proliferation and apoptosis. The messenger RNA (mRNA) and protein levels were assayed by RT-qPCR and western blotting, respectively. The targeted genes were predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. Cell proliferation was analyzed by CCK-8 assay. Annexin V-FITC/PI staining was used to evaluate the apoptosis in AGS and SGC-7901 cells. FGFR1 was frequently up-regulated in gastric cancer tissues and associated with poor overall survival in patients with gastric cancer. Interestingly, FGFR1 loss-of-function resulted in a significant growth inhibition and apoptosis in AGS and SGC-7901 cells. In addition, we found that miR-497 was inhibited in gastric cancer tissues and cell lines, while overexpression of miR-497 could suppress proliferation and induce apoptosis in AGS and SGC-7901 cells. Importantly, bioinformatics analysis and experimental data suggested that FGFR1 was a direct target of miR-497, which could inhibit FGFR1 expression when transfected with miR-497 mimics. Furthermore, we found that overexpression of FGFR1 reversed the growth inhibition and apoptosis of miR-497 mimics in AGS and SGC-7901 cells. These findings suggested that overexpression of miR-497 inhibited proliferation and induced apoptosis in gastric cancer through the suppression of FGFR1.
Subject(s)
Humans , Stomach Neoplasms/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/metabolism , Receptor, Fibroblast Growth Factor, Type 1/genetics , Prognosis , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Immunohistochemistry , Signal Transduction , Blotting, Western , Apoptosis , Disease Progression , Cell Line, Tumor , Cell Proliferation , Receptor, Fibroblast Growth Factor, Type 1/metabolism , Real-Time Polymerase Chain ReactionABSTRACT
In this study, we investigated the feasibility of using autologous vein graft and platelet-derived growth factors to bridge transected cavernous nerve in a rat model. A short defect in the bilateral cavernous nerve was created and repaired with vein graft from the right jugular vein or vein graft plus platelet-derived growth factors. The 32 rats were divided into four groups, namely Group 1 - no repair as a negative control, Group 2 - vein graft alone, Group 3 - vein graft plus platelet-derived growth factors, and Group 4 - sham operation as a positive control. We evaluated nerve regeneration and functional recovery using retrograde tracing study with FluoroGold, Toluidine blue staining of cavernous nerve, and the intracavernous pressure at 3 months. Three months after surgery, rich FluoroGold-positive cells were observed in the sham and vein graft plus platelet-derived growth factors group, but very few were found in the no repair group. The number of myelinated axons of regenerated cavernous nerve and intracavernous pressure were increased obviously in the two vein graft groups, especially in the vein graft plus platelet-derived growth factors group. These findings confirm the feasibility of using autologous vein as guides for cavernous nerve regeneration, and the regeneration can be further enhanced when the vein is filled with platelet-derived growth factors.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of estrogen receptor alpha (ERa) and insulin-like growth factor 1 (IGF1) on the proliferation of prostatic smooth muscle cells (PSMCs) in vitro.</p><p><b>METHODS</b>The ERalpha shRNA expression frame was subcloned to the pGSadeno adenovirus vector by homologous recombination technology to construct the pGSaaeno-ERalpha vector. After the mouse PSMCs were transfected in vitro by pGSaaeno-ERalpha, the mRNA and protein expression levels of ERalpha were detected by RT-PCR and Western blot respectively. The expression of IGF1 in the ERa-reduced cells was determined by Western blot 6 hours after treatment with 17beta-estradiol (E2) at 10(-8) mol/L. The post-transfection activity of estrogen or exogenous IGF1 in the proliferation of PSMCs was evaluated by MTT chlormetric analysis.</p><p><b>RESULTS</b>After treatment with E2, the proliferation of PSMCs and the expression of the IGF1 gene were significantly increased in the normal control group (P <0.05), but not obviously changed in the ERalpha-siRNA group (P> 0.05). And exogenous IGF1 failed to induce the proliferation of the ERalpha-reduced PSMCs.</p><p><b>CONCLUSION</b>E2 induces the expression of IGF1 via ERalpha, and IGFl, with the interaction of ERalpha, promotes the proliferation of PSMCs.</p>
Subject(s)
Animals , Male , Mice , Cell Proliferation , Cells, Cultured , Estradiol , Pharmacology , Estrogen Receptor alpha , Metabolism , Insulin-Like Growth Factor I , Metabolism , Myocytes, Smooth Muscle , Cell Biology , Prostate , Cell Biology , RNA, Messenger , GeneticsABSTRACT
The aim of this study was to investigate the effect of platelet-rich plasma (PRP) on cavernous nerve (CN) regeneration and functional status in a nerve-crush rat model. Twenty-four Sprague-Dawley male rats were randomly divided into three equal groups: eight had a sham operation, eight underwent bilateral nerve crushing with no further intervention and eight underwent bilateral nerve crushing with an immediate application of PRP on the site of injury. Erectile function was assessed by CN electrostimulation at 3 months and nerve regeneration was assessed by toluidine blue staining of CN and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining of penile tissue. Three months after surgery, in the group that underwent bilateral nerve crushing with no further intervention, the functional evaluation showed a lower mean maximal intracavernous pressure (ICP) and maximal ICP per mean arterial pressure (MAP) with CN stimulation than those in the sham group. In the group with an immediate application of PRP, the mean maximal ICP and maximal ICP/MAP were significantly higher than those in the injured control group. Histologically, the group with the application of PRP had more myelinated axons of CNs and more NADPH-diaphorase-positive nerve fibres than the injured control group but fewer than the sham group. These results show that the application of PRP to the site of CN-crush injury facilitates nerve regeneration and recovery of erectile function. Our research indicates that clinical application of PRP has potential repairing effect on CN and peripheral nerves.
Subject(s)
Animals , Male , Rats , Disease Models, Animal , Electric Stimulation , Erectile Dysfunction , Pathology , Therapeutics , NADPH Dehydrogenase , Metabolism , Nerve Regeneration , Physiology , Penile Erection , Physiology , Penis , Peripheral Nerve Injuries , Peripheral Nerves , Metabolism , Pathology , Platelet Transfusion , Platelet-Rich Plasma , Radiculopathy , Pathology , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To investigate the levels of pro-inflammatory cytokine IFN-gamma and anti-inflammatory cytokine TGF-beta1, in the expressed prostatic secretion (EPS) of men with chronic abacterial prostatitis and their clinical significance.</p><p><b>METHODS</b>The levels of IFN-gamma and TGF-beta1, in the EPS of 20 patients with inflammatory chronic pelvic pain syndrome (type III A), 20 patients with non-inflammatory chronic pelvic pain syndrome (type Ill B) and 10 healthy men were measured using enzyme-linked immunosorbent assay (ELISA). The results were analysed comparatively with NIH-chronic prostatitis symptom index (NIH-CPSI).</p><p><b>RESULTS</b>IFN--gamma and TGF-beta1 levels were higher in III ([14.92 +/- 7. 85)], [8477.50 +/- 4612.45] ng/L) and III B ([13.74 +/- 5.96], [7946.50 +/- 5044.06] ng/L) prostatitis patients than those in the controls ([7.47 +/- 1.49], [2462.50 +/- 985.31] ng/L), P < 0.05 and P < 0.001 respectively. There was no statistically significant difference in cytokine levels between III A and Il B prostatitis patients. No correlation was found between NIH-CPSI and cytokine levels, r = 0.02, P = 0.86, r = 0.31, P = 0.76.</p><p><b>CONCLUSION</b>IFN-gamma and TGF-beta1, play a very important role in the etiology of chronic abacterial prostatitis and can be the objective parameters in the diagnosis of chronic abacterial prostatitis.</p>