小儿肾母细胞瘤中SOCS-1和SOCS-3的表达及其临床意义

目的 探讨小儿肾母细胞瘤中细胞因子信号转导抑制因子-1(SOCS-1)和细胞因子信号转导抑制因子-3(SOCS-3)的表达,以及与小儿肾母细胞瘤不同病理类型之间的内在关系.方法 应用RT-PCR法及Western-Blot法测定10例小儿正常肾脏组织、30例小儿肾母细胞瘤及瘤旁组织中sOCS-1和SOCS-3的表达.结果 与正常小儿肾脏组织(mRNA表达:0.721±0.183、0.694±0.148,蛋白表达:0.746±0.168、0.652±0.183)相比,肾母细胞瘤组织中SOCS-1和SOCS-3 mRNA表达(0.156±0.027、0.103±0.021)及蛋白的表达(0.205±0.079、0.185±0.041)均明显降低(P＜0.01).在肿瘤不同病理类型之间SOCS-1和SOCS-3mRNA的表达(预后良好型:0.175±0.086、0.149±0.100,预后不良型:0.128±0.074、0.091±0.067)有差异(P＜0.05).瘤旁组织SOCS-1 和SOCS-3mRNA的表达(0.572±0.154、0.315±0.137)及蛋白的表达(0.628±0.183、0.439±0.137)与正常肾脏组织有差异(P＜0.05).结论 SOCS-1和SOCS-3的表达下降可能与小儿肾母细胞瘤的发生、发展密切相关,与病理分期也有一定联系.

Abstract：

Objective To investigate the expression levels of suppressor of cytokine signaling-1 and suppressor of cytokine signaling-3 in the Wilms' tumor and to study the inherent relationship between the different pathological types. Methods The expression levels of SOCS-1 and SOCS-3 were assessed by RT-PCR and Western-Blot in ten normal kidney tissues, thirty nephroblastoma specimens and adjacent tissues. Results Compared to the normal kidney tissues(mRNA expressions: 0.721±0.183、0.694±0.148,protein expressions:0.746±0.168、0.652±0.183), the expressions of mRNA (0.156±0.027、0.103±0.021)and protein(0.205±0.079、0.185±0.041)of the SOCS-1 and SOCS-3 in nephroblastoma were significantly lower (P＜0.01). There were statistically significant between mRNA expressions and the different pathological types(FH:0.175±0.086,0、149±0.100,UH:0. 128±0.074、0.091±0.067) (P＜0.05).The difference betweenneuroblastoma and the normal tissues in expressions of mRNA (0. 572 ± 0. 154、0. 315 ± 0. 137)and protein(0. 628± 0. 183、0. 439 ± 0. 137)of the SOCS-1 and SOCS-3 were statistically significant (P＜0. 05). Conclusions The low SOCS-1 and SOCS-3 expressions may have a role in the development and progress of nephroblastoma, and the different pathological types.     

叉头样转录因子3及转化生长因子β在小儿肾母细胞瘤中的表达和意义

目的 探讨叉头样转录因子3(FOXP3)及转化生长因子β(TGF-β)mRNA在小儿肾母细胞瘤中的表达和临床意义.方法 采用RT-PCR方法检测30例肾母细胞瘤组织、30例瘤旁组织和3例正常肾组织中FOXP3及TGF-βmRNA的表达情况,观察两者表达与临床分期的关系.结果 与瘤旁组织和正常组织比较,FOXP3及TGF-βmRNA在肿瘤组织中表达均上升,为0.706±0.107、0.667±0.121,差异具有统计学意义(P＜0.001);二者mRNA在Ⅰ期肿瘤中的表达强度为0.535±0.086、0.564±0.050均低于Ⅱ期的0.741±0.074、0.704±0.067和Ⅲ期的0.843±0.053、0.734±0.085,差异具有统计学意义(P＜0.001);FOXP3和TGF-βmRNA表达强度呈明显正相关(r=0.749,P＜0.001).结论 小儿肾母细胞瘤组织中,FOXP3及TGF-βmRNA表达增高,可能与肾母细胞瘤的发展和转移有关.

Abstract：

Objective To investigate the expression levels and the clinical significance of FOXP3and TGF-β in nephroblastoma. Methods The expression levels of FOXP3 and TGF-β were evaluated using RT-RCR in 30 nephroblastoma,30 adjacent tissues and 3 normal tissues. Results The expression levels of FOXP3 and TGF-β in nephroblastoma tissues were higher than adjacent and normal tissues(P＜0. 001). Compared to the stage Ⅱ and Ⅲ the difference of the expression levels of FOXP3and TGF-β in stage Ⅰ were significant in nephroblastoma tissues(P＜0. 001). There was a positive correlation between FOXP3 and TGF-β. Conclusions The expression levels of FOXP3 and TGF-β were higher in tumor tisssues, suggesting that it may be related to metastasis and progression of the nephroblastoma.     

小儿肾母细胞瘤中SOCS-1和SOCS-3的表达及其临床意义

Objective To investigate the expression levels of suppressor of cytokine signaling-1 and suppressor of cytokine signaling-3 in the Wilms' tumor and to study the inherent relationship between the different pathological types. Methods The expression levels of SOCS-1 and SOCS-3 were assessed by RT-PCR and Western-Blot in ten normal kidney tissues, thirty nephroblastoma specimens and adjacent tissues. Results Compared to the normal kidney tissues(mRNA expressions: 0.721±0.183、0.694±0.148,protein expressions:0.746±0.168、0.652±0.183), the expressions of mRNA (0.156±0.027、0.103±0.021)and protein(0.205±0.079、0.185±0.041)of the SOCS-1 and SOCS-3 in nephroblastoma were significantly lower (P＜0.01). There were statistically significant between mRNA expressions and the different pathological types(FH:0.175±0.086,0、149±0.100,UH:0. 128±0.074、0.091±0.067) (P＜0.05).The difference betweenneuroblastoma and the normal tissues in expressions of mRNA (0. 572 ± 0. 154、0. 315 ± 0. 137)and protein(0. 628± 0. 183、0. 439 ± 0. 137)of the SOCS-1 and SOCS-3 were statistically significant (P＜0. 05). Conclusions The low SOCS-1 and SOCS-3 expressions may have a role in the development and progress of nephroblastoma, and the different pathological types.     

High-mobility group box-1 and receptor for advanced glycation end products in preterm infants with brain injury

Background:High-mobility group box-1 (HMGB1) protein acts as an important pro-inflammatory mediator,which is capable of activating inflammation and tissue repair.HMGB1 can bind to its receptor such as advanced glycation end products (RAGE).RAGE,in turn,can promote the production of pro-inflammatory cytokines.Soluble RAGE (sRAGE) is a truncated form of the receptor comprising the extracellular domain of RAGE and can inhibit RAGE-activation.The objective of this study was to investigate whether HMGB1 and RAGE are involved in the development of brain injury in preterm infants.Methods:In total,108 infants ≤34 weeks gestation at birth were divided into 3 groups according to cranial altrasound scan:mild brain damage (n=33),severe brain damage (n=8) and no brain damage (n=67).All the placentas were submitted for pathologic evaluation.Histological chorioamnionitis (HCA) was defined as neutrophil infiltration of amniotic membranes,umbilical cord or chorionic plate.Expressions of HMGB1 and RAGE proteins were assessed by immunohistochemical analysis.The concentration of HMGB1 and sRAGE in umbilical cord blood were measured by enzyme-linked immunosorbent assay.Results:The frequency of HCA was 30.12％.HCA was associated with elevated concentrations of HMGB1 and decreased sRAGE in umbilical cord blood.The severe brain injury group demonstrated higher cord blood HMGB1 concentrations (P＜0.001) and lower sRAGE concentrations (P＜0.001) than both other groups.Brain injury in the premature infants was linked to intense staining for HMGB1/RAGE,particularly in inflammatory cells.Conclusions:Changes of cord blood HMGB1 and sRAGE of premature infants had direct relationship with the degree of inflammation and severity of brain damage.Monitoring sRAGE and HMGB1 levels may be helpful to predict intrauterine infection and brain injury in premature infants.     

裸鼠皮下移植人神经母细胞瘤模型的建立和HIF-1α及其相关蛋白的表达

目的 探讨应用人神经母细胞瘤细胞株建立裸鼠皮下神经母细胞瘤模型,测定缺氧诱导因子(hypoxia-inducible factor-1,HIF-1α)及其相关蛋白在该模型中的表达,了解HIF-1α及其相关蛋白在神经母细胞瘤发生发展中的作用,为神经母细胞瘤(neuroblastoma NB)的发病机制研究和诊治提供新的思路与方法.方法 应用KP-N-NS人肾上腺神经母细胞瘤细胞株建立模型.采用S-P免疫组织化学方法检测裸小鼠皮下移植瘤、手术切除的神经母细胞瘤肿瘤标本、正常人肾上腺髓质组织中HIF-1α、VEGF及TGF-α的表达情况.结合免疫组化结果对HIF-1α阳性组与HIF-1α阴性组裸鼠的体重变化、瘤体重量、存活天数进行比较,并对实验结果进行统计学处理.结果 4周后解剖结果证实成模率80%;HIF-1α、VEGF及TGF-α在肿瘤组织中的阳性表达率显著高于正常组织(P＜0.01),肿瘤组与阳性对照组HIF-1α、VEGF、TGF-α三种蛋白的阳性染色的积分光密度(integrating optical density,IOD)值明显高于阴性对照组(P＜0.01).HIF-1α在肿瘤组与阳性对照组中的表达与VEGF的表达成正相关(P＜0.05),与TGF-α的表达无明显相关性(P＞0.05);HIF-1α阳性组与HIF-1α阴性组体重变化、瘤体重量及存活天数比较,差异有统计学意义(P＜0.01);HIF-1α阳性组较HIF-1α阴性组裸鼠生存时间短(P＜0.05).结论 该模型用于研究人类神经母细胞瘤,造模时间短,成瘤率高,生物学行为与人类神经母细胞瘤接近.HIF-1α、VEGF及TGF-α蛋白的阳性表达与神经母细胞瘤的浸润及转移有关.HIF-1α与VEGF在神经母细胞瘤的发生发展过程中可能存在一定的协同作用.HIF-1α的高表达是预后不良的一个重要指标.

Abstract：

Objective To study the HIF-1αand its related proteins' expression in an athymic mouse human neuroblastoma xenograft model. Methods The was established by subcutaneous injection of human adrenal neuroblastoma cells. The expressions of HIF-1α, VEGF and TGF-α were detected by S-P immunohistochemical techniques. The tumor group was divided into HIF-1α positive group and the HIF-1α negative group according to the immunohistochemistry results. The body weight,tumor weight and survival days of the HIF-1αpositive mice and HIF-1αnegative mice were statistically analyzed. Results Four weeks later, significant neuroblastoma growth occurred in 40 mice (80%).The positive HIF-1α, VEGF and TGF-α rate of the tumor group was (73. 70 ± 10. 68) %, (69. 80 ±9. 91) %, and (71.43 ± 8. 52) 0% , respectively. It is significantly increased compared with the control group (P＜0. 01 ), which were (9. 67 ± 4. 53) %, (6. 80 ± 3. 40) %, and (6. 50 ± 4. 44) %, respectively. The integrated optical density (IOD) of HIF-1α, VEGF and TGF-α was 141.97 ± 7. 98,151.85 ±14. 35, and 139. 94 ± 4. 50 in tumor group, and 141.34 ± 6. 44, 144. 06 ± 8. 51 and 149. 00 ± 13. 63 in positive control group. They were significantly higher than those of the negative control group (P＜0. 01 ). In the tumor group and positive control group, the expression of HIF-1α was positively correlated with the expression of VEGF (x2 = 7. 778,r= 0. 504,P＜0. 01). However, no correlation between the expression of HIF-1α and TGF-α was found(x2 = 0. 208,r= 0. 934,P＞0. 05). The body weight,tumor weight and survival days of HIF-1α positive mice were significantly different with those of HIF-1α negative mice (P＜0. 01 ). The mice with positive HIF-1α expression had shorter survival time than the mice with negative HIF-1α (x2 = 19. 013,P＜0. 05). Conclusions The athymic mouse human neuroblastoma xenograft model is a good model to research the biology behavior of human neuroblastoma.The overexpression of HIF-1α, VEGF and TGF-αwere found in neuroblastoma which may play important roles in tumor invasion and metastasis. HIF-1αis synergistic with VEGF to promote carcinogenesis. The overexpression of HIF-1αis correlated with poor prognosis.     

Notch配体、受体在小儿胆道畸形肝组织中的表达及意义

Objective To investigate the expressions of Notch ligands and its receptors in the liver tissues of the pediatric patients with bile duct malformations. Methods Twenty three patients including 12 patients with biliary atresia and 11 with choledochal cyst were enrolled in this study. The patients' liver specimens were harvested during surgery. Immunohistochemistry and real-time fluorescent quantitative RT-PCR were performed to examine the expression of Notch ligands and its receptors in the liver tissues. Nine health liver tissues served as controls. The clinical data of these patients were also collected and analyzed. Results The expression of Jag1 significantly increased in the proliferating ductules of the patients with biliary atresia. Jag2 was negative in the portal area of all patients. The mRNA of Jag1 of the patients with bile duct malformations was higher than that of controls (P＜0. 01). No difference of the mRNA of Jag2 was found between the bile duct malformation patients and the controls (P＞0. 05). The expression of Notch 1 and Notch 2 was mostly found in hepatocytes and bile ductules of the controls' and the choledochal cyst patients' liver tissues. No Notch 1 and Notch 2 expression was found in proliferating ductules of the biliary atresia patients. Notch 3 was expressed in the neovascularization and mesenchyme of the biliary atresia patients. There was no significant difference of Notch1 and Notch 2 mRNA between the biliary atresia patients, choledochal cyst patients and controls (P＞0. 05). Notch 3 mRNA of biliary atresia patients was higher than that of the controls (P＜0. 01). Notch 4 expression was undetectable in all patients. Conclusions The expression patterns of Notch ligands and its receptors are changed in the patients with biliary atresia, which may contribute to the pathogenesis of biliary atresia.     

胎羊单侧输尿管不完全性梗阻后肾脏足细胞表型转化及PAX2、VEGF的表达

Objective To investigate the phenotypic change of the podocytes and paired box gene 2 (PAX2) and vascular endothelial growth factor (VEGF) expressions in fetal lambs' kidneys with unilateral partial ureteral obstruction. Methods Sixteen fetal lambs were randomly assigned to the obstruction and control group. The obstruction group included 12 lambs, whose superior segment ureters were tied with splited silastic tube to induce unilateral partial ureteral obstruction between 75and 85 gestational days. The control group had 4 fetal lambs subjected to sham operation. After birth,the kidneys of the lambs were harvested to study the phenotypic change of the podocytes as well as expressions of PAX2 and VEGF in the kidneys. Results Nine out of the 12 lambs of the obstruction group were delivered alive. All 4 sham operated lambs were delivered alive. Renal cortex cysts of varying sizes, interstitial fibrosis, a decrease in glomeruli number, and severe podocyte foot process fusion (4. 20± 1.08% vs 86. 79 ± 1.66%) were found in the obstructed kidneys compared with the control kidneys. The PAX2 expression in obstructed kidneys was significantly higher than that of the control kidneys (1.43 ± 0. 09 vs 2. 44 ± 0. 09, P＜0. 05). The VEGF expression was significantly decreased compared with the control kidneys (0. 80 ± 0. 15 vs 0. 33 ± 0. 14, P＜0. 05). Conclusions The changes of the phenotype and PAX2 and VEGF expressions may play roles in the pathogenesis of obstructive nephropathy.     

Dysregulated Hepatic Expression of Glucose Transporter Type-1,Toll-Like Receptor 4,and Nuclear Factor Kappa B in Estrogen-Induced Cholestasis Pregnant Rats with Placental Ischemia-Reperfusion Stress

Objective:This study aimed at investigating the expression of nuclear factor kappa B(NF-κB)and mammalian target of rapamycin(mTOR)related signal pathways in liver tissues of intrahepatic cholestasis of pregnancy animal models.Methods:Estrogen(EE)-induced cholestasis and a placental ischemia-reperfusion(IR)model were established in pregnant rats.All pregnant rats were divided into four groups by random number table:EE-IR group(n=6),EE-sham group(n=6),control-IR group(n=6)and control-sham group(n=6).Liver expression of mTOR,its upstream regulator DNA damage response-1(REDD1),and downstream factor glucose transporter type-1(GLUT1),accompanied by NF-κB(p65 is the most important component),its activator toll-like receptor 4(TLR4),and inhibitor IκBα,were detected by western blot analysis and real-time polymerase chain reaction.The intergroup comparisons were performed with a one-way analysis of variance,the comparisons among groups were analyzed with the nonparametric Kruskal-Wallis test.Results:Giving pregnant rats EE alone reduced the hepatic expression of IκBα(0.72±0.20vs.1.01±0.07,P=0.008).Meanwhile,giving pregnant rats placental IR alone increased liver levels of REDD1(3.24±0.98vs.1.06±0.24,P=0.025),GLUT1(2.37±0.82vs.1.09±0.10,P=0.039),TLR4(2.12±0.29vs.1.20±0.28,P=0.010),and p65(2.09±0.85vs.1.04±0.06,P=0.023),and decreased hepatic mTOR(0.50±0.07vs.1.01±0.03,P=0.001)and IκBα(0.61±0.08vs.1.01±0.07,P=0.014)expression.Subjecting EE-treated rats to placental IR did not further alter liver levels of GLUT1(2.02±0.45vs.1.79±0.39,P=0.240),TLR4(2.10±0.74vs.1.60±0.36,P=0.129),or p65(2.41±0.83vs.1.65±0.46,P=0.145),whereas it did decrease hepatic mTOR(0.42±0.09vs.0.90±0.14,P=0.008)and IκBα(0.43±0.09vs.0.72±0.20,P=0.004)expression and enhance REDD1 expression(4.46±0.65vs.2.05±0.47,P=0.009).Placental IR stress did impact the hepatic expression of REDD1-mTOR-GLUT1 and TLR4/NF-κB/IκBαin pregnant rats.Conclusion:Placental IR-induced hepatic GLUT1,TLR4,and p65 alternation,which responded efficiently in control rats,were impaired in EE-induced ICP rats.     

小儿肾母细胞瘤组织中VEGF-C的表达对微血管和淋巴管生成的影响及预后意义

目的 探讨小儿肾母细胞瘤组织中VEGF-C的表达对微血管、淋巴管生成的影响及其预后意义。方法 采用免疫组织化学PV9000法检测46例小儿肾母细胞瘤组织、19例瘤旁组织及8例正常肾组织中VEGF-C、CD34和LYVE-1的表达,并进行微血管密度(MVD)和淋巴管密度(LVD)计数。采用单因素分析VEGF-C、MVD和LVD与小儿肾母细胞瘤临床病理特征的关系;采用Kaplan-Meier法和Log-rank检验评价VEGF-C、MVD和LVD与患者预后的关系,并用Cox比例风险模型进行多因素分析。结果 在正常肾组织、瘤旁组织和肾母细胞瘤三种组织中,VEGF-C蛋白高表达率（12.50％、68.42％和73.91％）、MVD(8.25±2.82、13.32±3.94和16.98±3.74)和LVD(2.75±1.58、5.26±2.26和4.72±1.88)三者的表达差异均有统计学意义(P＜0.05);VEGF-C高表达、MVD和LVD与肿瘤临床分期、血管浸润、淋巴结转移有关(P＜0.05),MVD还与肿瘤的大小有关(P=0.029);VEGF-C表达、LVD和MVD三者间相互均有正相关关系(P＜0.05);Cox多因素分析显示,血管浸润、MVD和LVD是影响患儿生存的独立危险因素,化疗程数是影响患儿生存的独立保护因素。结论 VEGF-C高表达促进肾母细胞瘤微血管和淋巴管的生成,并可促进肿瘤转移而影响患儿预后。     

肾母细胞瘤组织中PTEN、PDCD4的表达及意义

目的 探究肾母细胞瘤组织中具有磷酸酶活性的抑癌基因PTEN及肿瘤抑制因子PDCD4 mRNA和蛋白表达及其临床病理联系。方法 采用RT-PCR和免疫组织化学法检测39例肾母细胞瘤和对照组15例癌旁2cm肾组织中PTEN、PDCD4 mRNA和蛋白表达,结合临床病理资料进行分析。结果 39例肾母细胞瘤组织中PTEN和PDCD4 mRNA均发生表达下调,且两种分子的mRNA的光密度在癌旁组织与肿瘤组织中比值均大于2。肾母细胞瘤组PTEN、PDCD4蛋白表达阳性率分别为17.95％、33.33％,远低于癌旁对照组阳性率93.33％、100％,差异有统计学意义(P＜0.05)。肾母细胞瘤组PTEN、PDCD4蛋白表达下调与患儿年龄、组织分级及临床分级、有无淋巴结转移等多种临床病理因素密切相关(P＜0.05)。肾母细胞瘤组与对照组PTEN、PDCD4蛋白表达在不同性别患儿中的差别不具有统计学意义(P＞0.05)。经直线相关分析,在肾母细胞瘤组织中PTEN与PDCD4蛋白表达存在正相关性(r= 0.877629,P＜0.05)。结论 肾母细胞瘤中PTEN及PDCD4 mRNA及蛋白与癌旁对照组织相比表达明显下调,可作为评价肾母细胞瘤生物学行为的参考指标。     

静脉肾盂造影不显影的肾母细胞瘤

对临床怀疑肾母细胞瘤病例结合静脉肾盂造影（ＩＶＰ）及病理检查进行对照研究。ＩＶＰ显影者占４４．８％，Ｘ线诊断为肾母细胞瘤中，有２例病理诊断分别为成神经细胞瘤和畸胎瘤；ｌ例ＩＶＰ无异常者，病理诊断为肾母细胞瘤。ＩＶＰ不显影者占５５．２％，病理诊断均为肾母细胞瘤。对于小儿腹部实质性肿块，如果ＩＶＰ不显影，仍应考虑有肾母细胞瘤可能。     

Wnt通路中相关基因表达在肾母细胞瘤发生中的作用

目的 研究 β-catenin、Wnt-4和sFRP-1基因在肾母细胞瘤、瘤旁肾组织、胚胎肾和正常肾组织中的表达实验对象分组,肾母细胞瘤组20例,瘤旁肾组织20例,胚胎肾组10例,正常肾组织8例.采用实时定量PCR的方法,分别检测上述各组标本中β-catenin、Wnt-4和sFRP-t基因的表达.结果 肾母细胞瘤组织、瘤旁肾组织和胚胎肾组织中β-catenin的表达量分别为4.6800±0.3086、4.5020±0.2385 和4.4960±0.3846,三组间差别无统计学意义.但均高于正常肾组织中β-catenin的表达量2.9220±0.3013,差别具有统计学意义.肾母细胞瘤组织Wnt一4的表达量3.7150±0.060明显高于瘤旁肾组织和胚胎肾组织中Wnt-4的表达(分别为3.1260±0.090和3.1250±0.073),前三者又均明显高于正常肾组织中Wnt-4的表达量2.6680±0.080.肾母细胞瘤组织和瘤旁肾组织中sFRP-1的表达量分别为4.6465±0.2890～f114.8020±0.2943,两组间差别无统计学意义.胚胎肾组织和正常肾组织中sFRP-1的表达量分别为2.7980 ±0.2003和2.9420±0.3867,两组间差别无统计学意义.但是肾母细胞瘤和瘤旁肾组织中sFRP-I的表达均明显高于胚胎肾组织和正常肾组织,差别具有高度的统计学意义.结论 Wnt信号通路中相关分子的表达异常可能是肾母细胞瘤发生的重要原因之一.     

TGFBR1和FGF9蛋白参与miRNA-140-5p调控肾母细胞瘤细胞的增殖过程

目的 本研究拟探讨rniRNA-140-5p在肾母细胞瘤增殖过程中的调控作用以及TGF-BR1和FGF9蛋白参与此过程的作用机制.方法 采用miRNA芯片技术分析肾母细胞瘤患儿肿瘤组织和瘤旁正常组织的miRNA表达谱,结合生物信息学预测和文献报道,选取表达差异显著的miR-NA-140-5p作为研究对象,扩大样本进行Real-time PCR验证;MTS方法检测miRNA-140-5p对细胞增殖率的影响;双荧光素酶实验验证miRNA-140-5p的直接下游靶基因;Western-blot法检测TGF-BR1蛋白表达;ELISA法检测细胞上清中FGF9蛋白表达.结果发现与瘤旁组织相比,miRNA-140-5p在肾母细胞瘤组织和瘤旁正常组织的相对表达量分别为1.69±0.83和6.37±3.25,肾母细胞瘤组miRNA-140-5p的表达量明显降低(P＜0.05).在肾母细胞瘤细胞株G401及SK-NEP-1中,发现过表达miRNA-140-5p可有效降低G401及SK-NEP-1的增殖率,将miRNA无关序列(miRNA-con)转染组细胞增殖率设为100％,与其相比,转染miRNA-140-5p的G401细胞组的增殖率为(87±15)％比(100±20)％,SK-NEP-1细胞组为(91±12)％比(100±9)％,组间比较,差异均有统计学意义(P值均＜0.05).利用生物信息学软件对miRNA-140-5p直接下游靶基因进行预测,结合肾母细胞瘤相关基因,选择TGFBR1和FGF9基因为miRNA-140-5p的直接下游靶基因;通过双荧光素酶实验证实miRNA-140-5p可直接靶向TGFBR1和FGF9基因.将miRNA-con转染组TGFBR1蛋白表达量设为1,与其相比,过表达miRNA-140-5p的G401及SK-NEP-1细胞内,TGFBR1蛋白表达量降低(G401细胞降为0.67;SK-NEP-1细胞降为0.87)(P＜0.05).与miRNA-con转染对照组相比,G401及SK-NEP-1细胞上清中FGF9蛋白表达量下降,G401组为(478.66±66.32) pg/ml比(535.64±78.53) pg/ml,SK-NEP-1细胞组为(486.57±71.01)pg/ml比(601.26±77.68) pg/ml,组间比较,差异均有统计学意义(P值均＜0.05).结论 患儿肾母细胞瘤中miRNA-140-5p表达量低于瘤旁正常组织,miRNA-140-5p影响肾母细胞瘤细胞的增殖,TGFBR1和FGF9蛋白可能参与了miRNA-140-5p调控肾母细胞瘤细胞增殖的过程.     

肾母细胞瘤VEGF及其受体Flk-1的表达及其临床意义

目的 探讨肾母细胞瘤组织中血管内皮生长因子(VEGF)及其受体(Flk-1)的表达和肿瘤微血管密度(MVD)的临床意义.方法 应用免疫组织化学(SABC)法检测33例肾母细胞瘤、瘤旁组织和6例正常肾组织中VEGF、Flk-1表达和MVD计数.观察并分析三个指标之间的相关性,同时分析其临床意义.结果 33例肿瘤组织VEGF阳性表达率81.8%,Flk-1阳性表达率69.7%,瘤旁组织VEGF阳性表达率9.1%,Flk-1阳性表达率6.1%,6例正常肾组织VEGF及Flk-1均为阴性表达,三者之问差异具有统计学意义.肿瘤组织MVD值29.7±12.4,瘤旁组织MVD值10.3±9.6,正常肾组织MVD值9.8±2.3.肿瘤组织的血管密度与后二者比较差异具有统计学意义.VEGF表达与Flk-1、MVD计数正相关,Flk-1与MVD表达正相关,VEGF、Flk-1表达在不同临床分期,及转归之问均具有显著性差异.结论 肾母细胞瘤中VEGF及其受体Flk-1呈高表达,血管生成增多,VEGF、Flk-1表达与MVD密切相关,提示VEGF及其受体Flk-1参与肾母细胞瘤的血管形成过程,促进血管生成,与预后相关.     

血管内皮生长因子在增生性血管瘤组织和血清中表达的意义

目的　探讨血管内皮生长因子 (VEGF)在增生性血管瘤组织、邻近组织和外周血中的表达水平和分布规律 ,进一步了解血管瘤的发生机制。方法　取增生性血管瘤患儿外周血清和术中瘤组织及其邻近组织标本 ,以相同年龄非相关患儿作对照 ,用酶联免疫法测定血清VEGF水平 ;对肿瘤组织及其相邻组织行cDNA合成和PCR扩增 ,提取VEGFmRNA ,并进行定量分析。结果　血管瘤患儿血清VEGF水平明显高于对照组 (P <0 .0 5 ) ;血管瘤组织的VEGFmRNA电泳阳性率为 10 0 % (9/ 9) ,邻近组织为 88.9% (8/ 9) ,对照组为 11.1% (1/9) ,经检验血管瘤及其邻近组织与对照组有显著性差异 (P均 <0 .0 1)。荧光定量测定mRNA在血管瘤及其邻近组织无显著差异 (P <0 .0 5 ) ,而两者与对照组均有显著差异 (P均 <0 .0 1)。结论　增生性血管瘤组织及患儿外周血VEGF明显增高 ,对血管内皮细胞增生和分化成血管瘤细胞有促进作用 ,检测血清VEGF水平有助于血管瘤分型和增生情况的监测