Differentiation Of Mesenchymal Stem Cells In Tumor Microenvironment & The Application Of Intravesical Prostatic Protrusion Measured By Ultrasound

OBJECTIVE To investigate the tropism of mesenchymal stem cells(MSCs) to tumor microenvironment and the feasibility of bone marrow mesenchymal stem cells differentiating into myofibroblast in vitro.METHODS 1ml bone marrow was taken from greater trochanter of male New Zealand rabbit and MSCs were obtained by density gradient centrifugation and cultured routinely;the surface markers were tested by flow cytometry.VX2 tumor was aseptically excised and primary cultured.The tropism of MSCs for control group, 30%and 50%VX2 conditioned medium were determined by using Transwell migration assay.MSCs were incubated in 30%VX2 conditioned medium for 7 or 14 days.The mRNA levels and protein expression ofα-SMA and Vimentin were measured by RT-PCR and Westernblot methods.RESULTS MSCs presented a spindle shape.The cultured MSCs were CD29(+), CD44(+),CD45(-),CD106(+).VX2 cells showed spindle or polygon shape.In the Transwell test,we found that the migrated cells appeared more in 30%VX2 conditioned medium group than other groups under microscope,which was further confirmed by the results of colorimetric assay.The mRNA levels and protein expression ofα-SMA and Vimentin both significantly increased in the 7 days group than the control group(P＜0.05),which further increasd in the 14 days group(P＜0.05).CONCLUSION MSCs show tropism for tumor microenvironment and can differentiate into myofibroblast in tumor microenvironment in vitro.SIGNIFICANCE This experiment suggests that MSCs could migrate to tumor and then differentiate into myofibroblast under tumor microenvironment,which might be a pathway of MSCs promoting the growth of tumor.It also suggestes that MSCs may be the precursors of the tumor/carcinoma associated myofibroblasts. OBJECTIVE To investigate the effect of mesenchymal stem cells(MSCs) in the process of tumor development and the possibility of MSCs differentiating into vascular endothelial cells in tumor microenvironment.METHODS 20 male New Zealand rabbits were randomly divided into two groups: test group and control group.MSCs were isolated and cultured by bone marrow cell adherence.The bladder tumor models were built by embedding vx2 mass in swelled bladder mucosa in all rabbits.One week later,4',6-diamidino-2-phenylindole labeling MSCs were transplanted into tumor tissue in test group(n=10).Culture medium was injected in the tumor tissue of control group(n=10).The maximum diameter of tumor mass was measured by ultrasound at 2,4 weeks after vx-2 tumor mass was embedded. All animals were sacrificed at 4 weeks.The double labeling immunofluorescence for CD146 was performed to reveal whether engrafted cells can differentiate into vascular endothelial cells.Vascular density was compared between two groups.VX2 cells were primary cultured also.MSCs were incubated in 30%VX2 conditioned medium for 7 or 14 days.The protein expression of CD146 were measured by Westernblot methods.RESULTS There was no significant difference in maximum diameter of tumor mass between two groups at 2 weeks(test group 0.77±0.15cm vs control group 0.71±0.15cm,P＞0.05).The maximum diameter appear larger in test group at 4weeks (test group 3.82±0.94cm vs control group 2.28±0.54cm,P＜0.05). Immunofluorescence studies revealed some engrafted MSCs expressing a vascular endothelial cell phenotype(CD146).Furthermore,vascular density was augmented in test group compared with control group(10.1±0.70/0.2mm~2 vs 8.24±0.81/0.2mm~2, P＜0.05).In vitro,the protein expression of CD146 significantly increased in the 7 days group than the control group(P＜0.05),which further increasd in the 14 days group(P＜0.05).CONCLUSION Engrafted MSCs can differentiate into vascular endothelial cells and contribute to angiogenesis in tumor microenvironment,which may be the major pathway of promoting tumor growth. OBJECTIVE To evaluate a noninvasive method to predict bladder outlet obstruction(BOO) and bladder function in patients with benign prostatic enlargement (BPE) based on intravesical prostatic protrusion(IPP) using transabdominal ultrasound.METHODS The records of 206 first visit patients with BPE were analyzed. Patients were divided into two groups based on the degree of IPP:the significant IPP group-greater than 10 mm and no significant IPP group-10 mm or less.Clinical data and urodynamic findings of the two groups were analyzed to define the clinical significance of IPP.RESULTS Increased prostate volume(73.7±35.9cm~3 vs 62.8±36.5cm~3),serum prostate specific antigen(1.81±0.67ng/ml vs 1.64±0.36ng/ml),post-voiding residual urine volume(290.2±217.2ml vs 228.2±167.9ml),incidence of acute urine residual (33.3%vs 18.0%) and bladder trabeculation(23.1%vs 11.7%) appeared more often in the significant IPP group(P＜0.05).In the urodynamic findings,significantly lower peak flow rate(Qmax)(7.6±4.1 ml/s vs 9.1±3.6ml/s ) and higher incidence of detrusor overactivity(82.1%vs 17.2%) and low bladder compliance(35.9%vs 12.5%) both existed in the significant IPP group(P＜0.01).In addition,maximum detrusor pressure (109.8±84.9cmH20 vs 84.9±44.1 cmH2O) and BOO index(75.2±27.1 vs 65.9±34.6) were significantly higher in the significant IPP group(p＜0.05).The incidence of recurrence of acute urinary intention was higher in the significant IPP group(64.3% vs 23.5%)(P＜0.05).CONCLUSION IPP is a useful predictor for evaluating BOO and detrusor function. BOO and impaired detrusor function in the significant IPP patients are more severe. The significant IPP patients,especially those presenting with AUR,may benefit from early surgical intervention.