New stem cell based therapies are undergoing intense research and are widely investigated in clinical fields including the urinary system. The urinary bladder performs critical complex functions that rely on its highl...New stem cell based therapies are undergoing intense research and are widely investigated in clinical fields including the urinary system. The urinary bladder performs critical complex functions that rely on its highly coordinated anatomical composition and multiplex of regulatory mechanisms. Bladder pathologies resulting in severe dysfunction are common clinical encounter and often cause significant impairment of patient's quality of life. Current surgical and medical interventions to correct urinary dysfunction or to replace an absent or defective bladder are sub-optimal and are associated with notable complications. As a result, stem cell based therapies for the urinary bladder are hoped to offer new venues that could make up for limitations of existing therapies. In this article, we review research efforts that describe the use of different types of stem cells in bladder reconstruction, urinary incontinence and retention disorders. In particular, stress urinary incontinence has been a popular target for stem cell based therapies in reported clinical trials. Furthermore, we discuss the relevance of the cancer stem cell hypothesis to the development of bladder cancer. A key subject that should not be overlooked is the safety and quality of stem cell based therapies introduced to human subjects either in a research or a clinical context.展开更多
Purpose: The main objectives of this study were to characterize and compare the urothelial stem cells (healthy and cancer cells) and TLRs features in the urinary bladder of men without lesionsand with non-muscle-invas...Purpose: The main objectives of this study were to characterize and compare the urothelial stem cells (healthy and cancer cells) and TLRs features in the urinary bladder of men without lesionsand with non-muscle-invasive and muscle invasive urothelial tumors. Materials and Methods: Thirty samples of the urinary bladder of 50 to 80-year-old men, with and without diagnosis of malignant urothelial lesions were used. The 30 samples were divided into 3 groups (n = 10 per group): Normal Group;Non-Muscle Invasive Bladder Cancer Group;Muscle Invasive Bladder Cancer Group. The samples were histopathologically and immunohistochemically analyzed. The study was conducted at teaching Hospital of the University of Campinas (UNICAMP). Results: The CD44 and CD133 immunoreactivities were significantly intense in the muscle-invasive cancer group when compared to the other groups. The ABCG2 biomarker demonstrated intense immunoreactivities in both non-muscle and muscle invasive groups, and absent immunoreactivity in the normal group. All groups showed weak CD117 immunoreactivity. Putative Healthy Stem Cells (CD44/CD133/ CD117+) occurred in all groups. Putative Cancer Stem Cells (CD44/CD133/ABCG2+) only occurred in the non-muscle and muscle invasive cancer groups. TLR2 immunoreactivity was significantly lower in the non-muscle invasive cancer group and absent in the muscle invasive cancer group. TLR4 immunoreactivity was significantly lower in both cancer groups. Conclusions: This study leads us to the conclusion that putative cancer stem cell occurrence was sensitive to the decreased in TLR2 and TLR4 immunoreactivities. Also, TLR2 and TLR4 demonstrated their involvement in the regulation of the different biomarkers for putative healthy and cancer urothelial stem cells, probably acting as negative regulators of urothelial carcinogenesis. Taken together data obtained suggest that use of TLRs agonists could be a promising alternative for the treatment of non-muscle and muscle invasive bladder tumors.展开更多
In order to detect the expression of Oct4 in bladder cancer tissue and cell line BIU-87, immunohistochemistry was used in 49 bladder cancer biopsy samples and immunofluorescence and reverse transcription-PCR were perf...In order to detect the expression of Oct4 in bladder cancer tissue and cell line BIU-87, immunohistochemistry was used in 49 bladder cancer biopsy samples and immunofluorescence and reverse transcription-PCR were performed on bladder cancer cell line BIU-87. Forty of 49 bladder cancer samples showed the expression of Oct4 in about 0.6% cancer cells, The positive rate and density of Oct4 expression had no obvious relationship with the grade, recurrence or metastasis of bladder cancer （P〉0.05）. A few Oct4 positive cells were found in bladder cancer cell line BIU-87, which was also confirmed by RT-PCR. This study indicated the existence of few Oct4 positive cells in bladder cancer, which may be the bladder cancer stem cells. This study may provide the foundation for isolation and identification of bladder cancer stem cells.展开更多
Urothelial carcinoma of bladder is the second most prevalent genitourinary disease.It is a highly heterogeneous disease as it represents a spectrum of neoplasms,including non-muscle invasive bladder cancer(NMIBC),musc...Urothelial carcinoma of bladder is the second most prevalent genitourinary disease.It is a highly heterogeneous disease as it represents a spectrum of neoplasms,including non-muscle invasive bladder cancer(NMIBC),muscle invasive bladder cancer(MIBC)and metastatic lesions.Genome-wide approaches and candidate gene analysis suggest that malignant transformation of the bladder is multifactorial and a multitude of genes are involved in the development of MIBC or NMIBC phenotypes.Wnt signaling is being examined to control and maintain balance between stemness and differentiation in adult stem cell niches.Owing to its participation in urothelial development and maintenance of adult urothelial tissue homeostasis,the components of Wnt signaling are reported as an important diagnostic and prognostic markers as well as novel therapeutic targets.Mutations/epigenetic alterations in the key molecules of Wnt/β-catenin canonical pathway have been linked with tumorigenesis,development of drug resistance and enhanced survival.Present review extends our understanding on the functions of key regulatory molecules of canonical Wnt/β-catenin pathway in urothelial tumorigenesis by inducing cancer stem cell phenotype(UCSCs).UCSCs may be responsible for tumor heterogeneity,high recurrence rates and complex biological behavior of bladder cancer.Therefore,understanding the role of UCSCs and the regulatory mechanisms that are responsible for high relapse rates and metastasis could help to develop pathway inhibitors and augment current therapies.Potential implications in the treatment of urothelial carcinoma of bladder by targeting this pathway primarily in UCSCs as well as in bulk tumor population that are responsible for high relapse rates and metastasis may facilitate potential therapeutic avenues and better prognosis.展开更多
目的:研究肿瘤相关抗原,黑色素瘤抗原家族A成员3(melanoma antigen family A,3;MAGE-A3),在人类膀胱癌干细胞中的表达情况,并探讨其意义。方法:采用反转录聚合酶链反应(RT-PCR)技术和Western blot技术检测MAGE-A3在人类膀胱癌细胞系T24...目的:研究肿瘤相关抗原,黑色素瘤抗原家族A成员3(melanoma antigen family A,3;MAGE-A3),在人类膀胱癌干细胞中的表达情况,并探讨其意义。方法:采用反转录聚合酶链反应(RT-PCR)技术和Western blot技术检测MAGE-A3在人类膀胱癌细胞系T24细胞中及从其中分离出来的具有干细胞特性的T24侧群细胞中的表达情况;采用免疫荧光双标记技术检测MAGE-A3和膀胱癌干细胞的一个标志物CD133在T24侧群细胞中的共表达情况。结果:在mRNA和蛋白水平,MAGE-A3在具有癌干细胞特性的T24侧群细胞中的表达水平明显高于对应的母系T24细胞;MAGE-A3和膀胱癌干细胞标志物CD133在T24侧群细胞中有阳性共表达。结论:MAGE-A3在人类膀胱癌干细胞中有特异性的高表达,有望成为膀胱癌干细胞一个新的、特异性的标志物,及针对膀胱癌干细胞进行免疫治疗的一个新的靶点。展开更多
目的探讨线粒体核糖体蛋白S5(mitochondrial ribosomal protein S5,MRPS5)对人膀胱癌细胞的增殖能力及其中干细胞干性特征的影响。方法 Western blot检测人膀胱癌及癌旁正常组织MRPS5的表达;构建靶向干扰MRPS5的慢病毒载体,感染膀胱癌...目的探讨线粒体核糖体蛋白S5(mitochondrial ribosomal protein S5,MRPS5)对人膀胱癌细胞的增殖能力及其中干细胞干性特征的影响。方法 Western blot检测人膀胱癌及癌旁正常组织MRPS5的表达;构建靶向干扰MRPS5的慢病毒载体,感染膀胱癌细胞株T24,以干扰Scramble序列作为对照,Western blot检测MRPS5干扰效率;细胞活力实验检测细胞增殖能力;流式细胞术检测细胞周期;细胞成球实验观察成球能力;Western blot检测肿瘤干性转录因子Nanog,Oct4,c-Myc和Sox2的表达;小鼠皮下移植瘤实验观察T24体内成瘤能力。结果 MRPS5在膀胱癌组织中的表达高于癌旁正常组织;慢病毒干扰载体PLKO.1-sh MRPS5有效,且干扰MRPS5表达后T24细胞增殖能力下降(P<0.05),周期阻滞于S期,干性因子表达均下调,成球率无变化(P>0.05)但成球直径明显减小;同时小鼠体内成瘤体积减小[(0.784±0.278)vs(0.500±0.245)cm3,P<0.05],瘤质量减轻[(0.862±0.372)vs(0.412±0.248)g,P<0.05]。结论 MRPS5在膀胱癌中较高表达,且干扰MRPS5表达能抑制T24增殖并抑制T24中的干细胞生物学特征。展开更多
基金funding from the Science Technology Development Fund (STDF), Egypt
文摘New stem cell based therapies are undergoing intense research and are widely investigated in clinical fields including the urinary system. The urinary bladder performs critical complex functions that rely on its highly coordinated anatomical composition and multiplex of regulatory mechanisms. Bladder pathologies resulting in severe dysfunction are common clinical encounter and often cause significant impairment of patient's quality of life. Current surgical and medical interventions to correct urinary dysfunction or to replace an absent or defective bladder are sub-optimal and are associated with notable complications. As a result, stem cell based therapies for the urinary bladder are hoped to offer new venues that could make up for limitations of existing therapies. In this article, we review research efforts that describe the use of different types of stem cells in bladder reconstruction, urinary incontinence and retention disorders. In particular, stress urinary incontinence has been a popular target for stem cell based therapies in reported clinical trials. Furthermore, we discuss the relevance of the cancer stem cell hypothesis to the development of bladder cancer. A key subject that should not be overlooked is the safety and quality of stem cell based therapies introduced to human subjects either in a research or a clinical context.
文摘Purpose: The main objectives of this study were to characterize and compare the urothelial stem cells (healthy and cancer cells) and TLRs features in the urinary bladder of men without lesionsand with non-muscle-invasive and muscle invasive urothelial tumors. Materials and Methods: Thirty samples of the urinary bladder of 50 to 80-year-old men, with and without diagnosis of malignant urothelial lesions were used. The 30 samples were divided into 3 groups (n = 10 per group): Normal Group;Non-Muscle Invasive Bladder Cancer Group;Muscle Invasive Bladder Cancer Group. The samples were histopathologically and immunohistochemically analyzed. The study was conducted at teaching Hospital of the University of Campinas (UNICAMP). Results: The CD44 and CD133 immunoreactivities were significantly intense in the muscle-invasive cancer group when compared to the other groups. The ABCG2 biomarker demonstrated intense immunoreactivities in both non-muscle and muscle invasive groups, and absent immunoreactivity in the normal group. All groups showed weak CD117 immunoreactivity. Putative Healthy Stem Cells (CD44/CD133/ CD117+) occurred in all groups. Putative Cancer Stem Cells (CD44/CD133/ABCG2+) only occurred in the non-muscle and muscle invasive cancer groups. TLR2 immunoreactivity was significantly lower in the non-muscle invasive cancer group and absent in the muscle invasive cancer group. TLR4 immunoreactivity was significantly lower in both cancer groups. Conclusions: This study leads us to the conclusion that putative cancer stem cell occurrence was sensitive to the decreased in TLR2 and TLR4 immunoreactivities. Also, TLR2 and TLR4 demonstrated their involvement in the regulation of the different biomarkers for putative healthy and cancer urothelial stem cells, probably acting as negative regulators of urothelial carcinogenesis. Taken together data obtained suggest that use of TLRs agonists could be a promising alternative for the treatment of non-muscle and muscle invasive bladder tumors.
文摘In order to detect the expression of Oct4 in bladder cancer tissue and cell line BIU-87, immunohistochemistry was used in 49 bladder cancer biopsy samples and immunofluorescence and reverse transcription-PCR were performed on bladder cancer cell line BIU-87. Forty of 49 bladder cancer samples showed the expression of Oct4 in about 0.6% cancer cells, The positive rate and density of Oct4 expression had no obvious relationship with the grade, recurrence or metastasis of bladder cancer （P〉0.05）. A few Oct4 positive cells were found in bladder cancer cell line BIU-87, which was also confirmed by RT-PCR. This study indicated the existence of few Oct4 positive cells in bladder cancer, which may be the bladder cancer stem cells. This study may provide the foundation for isolation and identification of bladder cancer stem cells.
基金University Grants Commission, Govt. of India for providing financial support
文摘Urothelial carcinoma of bladder is the second most prevalent genitourinary disease.It is a highly heterogeneous disease as it represents a spectrum of neoplasms,including non-muscle invasive bladder cancer(NMIBC),muscle invasive bladder cancer(MIBC)and metastatic lesions.Genome-wide approaches and candidate gene analysis suggest that malignant transformation of the bladder is multifactorial and a multitude of genes are involved in the development of MIBC or NMIBC phenotypes.Wnt signaling is being examined to control and maintain balance between stemness and differentiation in adult stem cell niches.Owing to its participation in urothelial development and maintenance of adult urothelial tissue homeostasis,the components of Wnt signaling are reported as an important diagnostic and prognostic markers as well as novel therapeutic targets.Mutations/epigenetic alterations in the key molecules of Wnt/β-catenin canonical pathway have been linked with tumorigenesis,development of drug resistance and enhanced survival.Present review extends our understanding on the functions of key regulatory molecules of canonical Wnt/β-catenin pathway in urothelial tumorigenesis by inducing cancer stem cell phenotype(UCSCs).UCSCs may be responsible for tumor heterogeneity,high recurrence rates and complex biological behavior of bladder cancer.Therefore,understanding the role of UCSCs and the regulatory mechanisms that are responsible for high relapse rates and metastasis could help to develop pathway inhibitors and augment current therapies.Potential implications in the treatment of urothelial carcinoma of bladder by targeting this pathway primarily in UCSCs as well as in bulk tumor population that are responsible for high relapse rates and metastasis may facilitate potential therapeutic avenues and better prognosis.
文摘目的:研究肿瘤相关抗原,黑色素瘤抗原家族A成员3(melanoma antigen family A,3;MAGE-A3),在人类膀胱癌干细胞中的表达情况,并探讨其意义。方法:采用反转录聚合酶链反应(RT-PCR)技术和Western blot技术检测MAGE-A3在人类膀胱癌细胞系T24细胞中及从其中分离出来的具有干细胞特性的T24侧群细胞中的表达情况;采用免疫荧光双标记技术检测MAGE-A3和膀胱癌干细胞的一个标志物CD133在T24侧群细胞中的共表达情况。结果:在mRNA和蛋白水平,MAGE-A3在具有癌干细胞特性的T24侧群细胞中的表达水平明显高于对应的母系T24细胞;MAGE-A3和膀胱癌干细胞标志物CD133在T24侧群细胞中有阳性共表达。结论:MAGE-A3在人类膀胱癌干细胞中有特异性的高表达,有望成为膀胱癌干细胞一个新的、特异性的标志物,及针对膀胱癌干细胞进行免疫治疗的一个新的靶点。
