MTS1失活与膀胱癌发生的关系研究进展

多肿瘤抑制基因１（ＭＴＳ１）位于９ｐ２１，编码一个１６ＫＤ的蛋白质（ｐ１６），ｐ１６蛋白是周期素依赖性激酶４（ＣＤＫ４）的抑制因子，能抑制细胞的生长和分裂。ＭＴＳ１主要通过缺失、点突变和５’ＣｐＧ岛异常甲基化而失活，已经证实该基因的失活与多种组织的肿瘤形成有关，其中包括膀胱癌。本文就有关ＭＴＳ１失活与膀胱癌发生的研究进展作一综述。     

MTS1基因在人BPH中突变缺失的初步研究

目的探讨ＭＴＳ１基因在人前列腺增生症（ＢＰＨ）发病机制中的作用和变异情况。方法用ＰＣＲ银染ＳＳＣＰ技术检测１８例正常前列腺和３８例ＢＰＨ中抑癌基因ＭＴＳ１各外显子的纯合性缺失和突变。结果１２例ＢＰＨ出现ＭＴＳ１基因缺失,总缺失率为３２％;正常前列腺组织中有１例出现ＭＴＳ１基因缺失,缺失率为６％,两者之间差异有显著性（Ｐ＜００５）;无１例有ＭＴＳ１基因的突变。结论ＢＰＨ的发病机制可能与ＭＴＳ１的纯合性缺失有关,未见有突变发生。应对ＭＴＳ１基因在人ＢＰＨ中的作用作更深入的研究     

MTS1失活与膀胱癌发生的关系研究进展

多肿瘤抑制基因１（ＭＴＳ１）位于９ｐ２１，编码一个１６ＫＤ的蛋白质（ｐ１６），ｐ１６蛋白是周期素依赖性激酶４（ＣＤＫ４）的抑制因子，能抑制细胞的生长和分裂。ＭＴＳ１主要通过缺失、点突变和５’ＣｐＧ岛异常甲基化而失活，已经证实该基因的失活与多种组织的肿瘤形成有关，其中包括膀胱癌。本文就有关ＭＴＳ１失活与膀胱癌发生的研究进展作一综述。     

原发性膀胱癌多肿瘤抑制基因1改变及其意义

为探讨多肿瘤抑制基因１（ＭＴＳ１）在原发性膀胱癌发生发展中的作用，采用ＰＣＲＳＳＣＰ和ＤＮＡ直接测序方法，对１８例膀胱移行细胞癌进行ＭＴＳ１分析。结果１８例原发性膀胱癌中没有发现ＭＴＳ１纯合性缺失，但发现２例点突变。表明原发性膀胱癌的发生与ＭＴＳ１的失活相关     

直肠癌癌旁移行粘膜MTS1基因产物表达的意义

目的 探讨直肠癌癌旁移行粘膜（ＴＭ）的性质及多种种瘤抑制基因（ＭＴＳ１）基因缺失的临床意义。方法 应用粘液组化方法观察了直肠癌旁ＴＭ的范围,免疫组化方法观察了ＭＴＳ１基因产物表达情况,并与正常粘膜及癌组织进行对照 结果 ＭＴＳ１基因产物在正常直肠膜阳性表达率最高,在癌旁ＴＭ、癌组织表达逐渐减少,三者差异有显著意义（Ｐ〈０．０５）;ＭＴＳ１阴性的直肠癌病例,癌旁ＴＭ的明显高于ＭＴＳ１阳性的病例（Ｐ〉     

MTS1基因在膀胱癌中存在状态的研究

目的 观察人膀胱癌中ＭＴＳ１基因的存在状态。方法 运用双重对照ＰＣＲ及单链构象多态分析技术检测１７例原发性膀胱癌。结果 １７例膀胱癌中，２例表现为ＭＴＳ１基因纯合性缺失，２例表现为杂合性缺失，缺失频率为２３．５％（４／１７）未见异常泳动带，结论 ＭＴＳ１基因缺失与原发性膀胱癌的发生发展有关。     

MTS1逆转录病毒表达载体的构建与鉴定

ＭＴＳ１逆转录病毒表达载体的构建与鉴定袁建林，柴玉波，于茂生，惠宏襄多肿瘤抑制基因１（ＭｕｌｔｉｐｌｅＴｕｍｏｒＳｕｐｐｒｅｓｓＯｒ１，ＭＴＳ１）是１９９４年４月由美国盐湖城的Ｋａｍｂ与圣地亚哥的Ｎｏｂｏｒｌ首先报道的一种新型抑癌基因，其编码蛋白的分...     

MTS1基因产物P16蛋白在膀胱移行细胞癌中的表达

ＭＴＳ１基因产物Ｐ１６蛋白在膀胱移行细胞癌中的表达陈强王新生董胜国李玉军孙显露申东亮ＭＴＳ１（ｍｕｌｔｉｐｌｅｔｕｍｏｒｓｕｐｐｒｅｓｓｏｒ１）因子是新近发现的一种多肿瘤抑制基因，能负性调节细胞周期，并与多种人类肿瘤的发生、发展密切相关［１］。为探讨...     

大肠癌组织中MTS1／p16基因产物的表达

探讨ＭＴＳ１／ｐ１６基因失活与大肠癌发生的关系。方法 应用免疫组化方法观察了ＭＴＳ１／ｐ１６基因产物在１１０例大鼠癌组织的表达情况，并与非典型增生，部旁移行粘膜及正常粘膜进行了比较。结果 癌组织ｐ１６阳性分级表达低于正常粘膜及移行粘膜，粘膜液癌低于管状腺癌。     

P16基因与肺癌

ｐ１６基因与肺癌田辉陈明菊张超综述王善政审校肺癌是最常见的恶性肿瘤之一。随着分子生物学和基因工程的迅速发展，对肺癌的病因、演变和生长过程的研究已深入到分子水平。近年研究又发现了一个重要的新型抑癌基因，即ｐ１６基因又称ＭＴＳ１（ｍｕｌｔｉｐｌｅｔｕｍｏ...     

Growth Plate Borderline Chondrocytes Behave as Transient Mesenchymal Precursor Cells

The growth plate provides a substantial source of mesenchymal cells in the endosteal marrow space during endochondral ossification. The current model postulates that a group of chondrocytes in the hypertrophic zone can escape from apoptosis and transform into cells that eventually become osteoblasts in an area beneath the growth plate. The growth plate is composed of cells with various morphologies; particularly at the periphery of the growth plate immediately adjacent to the perichondrium are “borderline” chondrocytes, which align perpendicularly to other chondrocytes. However, in vivo cell fates of these special chondrocytes have not been revealed. Here we show that borderline chondrocytes in growth plates behave as transient mesenchymal precursor cells for osteoblasts and marrow stromal cells. A single-cell RNA-seq analysis revealed subpopulations of Col2a1-creER-marked neonatal chondrocytes and their cell type–specific markers. A tamoxifen pulse to Pthrp-creER mice in the neonatal stage (before the resting zone was formed) preferentially marked borderline chondrocytes. Following the chase, these cells marched into the nascent marrow space, expanded in the metaphyseal marrow, and became Col(2.3 kb)-GFP⁺ osteoblasts and Cxcl12-GFP^high reticular stromal “CAR” cells. Interestingly, these borderline chondrocyte-derived marrow cells were short-lived, as they were significantly reduced during adulthood. These findings demonstrate based on in vivo lineage-tracing experiments that borderline chondrocytes in the peripheral growth plate are a particularly important route for producing osteoblasts and marrow stromal cells in growing murine endochondral bones. A special microenvironment neighboring the osteogenic perichondrium might endow these chondrocytes with an enhanced potential to differentiate into marrow mesenchymal cells. © 2019 American Society for Bone and Mineral Research.     

Fibroblast growth factor 2 regulation of mitral valve interstitial cell repair in vitro

OBJECTIVE: Because elongated mitral valve interstitial cells have features of myofibroblasts, it is likely that these cells are essential for the repair of injured valve leaflets. We characterized the cellular morphology and pattern of repair of these interstitial cells in wounds produced in vitro and tested the hypothesis that fibroblast growth factor 2 enhances interstitial cell repair. METHODS: Mitral valve interstitial cells were plated onto glass coverslips, reached confluence after 1 week, and were wounded by passage of a spatula along the center of a monolayer, which created a linear wound with two edges. The wounds were observed from 0 to 96 hours by phase-contrast microscopy. Wounds were also fixed at 0, 2, and 24 hours and stained for fibroblast growth factor 2 and fibroblast growth factor receptor 1 by means of immunofluorescence and laser confocal microscopy. RESULTS: Cells in confluent monolayers and in the monolayer behind the wound edge formed a multilayered orthogonal pattern of elongated cells similar to fibroblasts. Cells along the wound edge migrated into the wound after 4 hours, and at 24 hours single cells with prominent lamellipodia and tails were present within the wound. There was overlapping of cells as well, similar to smooth muscle cells. Fibroblast growth factor 2 and fibroblast growth factor receptor 1 were present in the cells of the undisturbed confluent monolayer. They were upwardly regulated relative to the unwounded monolayer in the cells along the wound edge at 2 hours and in the monolayer behind the wound edge at 24 hours. In single cells that migrated into the wound, fibroblast growth factor 2 and fibroblast growth factor receptor 1 were prominent. Fibroblast growth factor 2 showed a 6-fold increase in concentration relative to unwounded cultures in conditioned medium from wounded cultures at 2 hours after wounding. Addition of a neutralizing antibody to fibroblast growth factor 2 significantly delayed wound closure at 24 to 96 hours. Addition of exogenous fibroblast growth factor 2 to cultures at the time of wounding did not enhance wound repair. CONCLUSION: Mitral valve interstitial cells have the ability to repair wounds, and fibroblast growth factor 2 is a modulator of these repair processes.     

体外培养版纳微型猪近交系骨髓基质干细胞的生物学特性

目的　分离培养成年版纳微型猪近交系骨髓基质干细胞并探讨其体外生物学特性。方法　成年版纳微型猪骨髓基质干细胞通过密度梯度离心法分离获得 ,用含 15 %小牛血清的 DMEM在 37℃ ,5 % CO2 条件下培养 ;通过克隆生长特性、特异抗原表达和成骨分化能力鉴定其干细胞特性。结果　版纳微型猪近交系骨髓分离获得的基质干细胞具有多种形状 ,但主要为梭形。其具有很强的增殖能力 ,在体外培养条件下可以观察到明显的克隆样生长 ;表达 SH2、SH3、SH4、SB10和 SB2 1等骨髓基质干细胞的特异标记 ;经成骨添加剂诱导培养之后能分化为成骨细胞 ,碱性磷酸酶活性增强并具有细胞外基质矿化能力。结论　成年版纳微型猪近交系骨髓来源的基质干细胞具有干细胞的一般生物学特性     

Adult kidney tubular cell population showing phenotypic plasticity, tubulogenic capacity, and integration capability into developing kidney

Using in vivo bromodeoxyuridine (BrdU) labeling, a tubular cell population (label-retaining tubular cells [LRTC]) was identified recently in normal adult kidneys, which contributes actively to the regeneration process of the kidney after injury. Here, these LRTC are characterized in vitro. The LRTC population was isolated from BrdU-treated rat kidney by FACS. Both LRTC and non-LRTC underwent proliferation and maintained an epithelial phenotype in the presence of tubulogenic growth factors such as EGF, TGF-alpha, IGF-I, and hepatocyte growth factor. It is interesting that LRTC also proliferated without epithelial markers expression in the presence of soluble factors derived from an embryonic kidney metanephric mesenchyme cell line. The type of extracellular matrix strongly influenced the phenotype of LRTC. Furthermore, in three-dimensional collagen gel culture, LRTC formed tubule-like or tubulocystic structures in response to growth factors (hepatocyte growth factor and fibroblast growth factor) that are known to induce kidney cell tubulogenesis in vitro and/or participate in renal regeneration in vivo. In contrast, non-LRTC did not form these structures. When transplanted into the metanephric kidney, LRTC but not non-LRTC were integrated into epithelial components of nephron, including the proximal tubular cells and the ureteric bud. They also differentiated into fibroblast-like cells. Collectively, these findings suggest that LRTC are an adult kidney tubular cell population that shows phenotypic plasticity, tubulogenic capacity, and integration capability into the developing kidney.     

Transforming growth factor-α with insulin induces proliferation in rat utricular extrasensory epithelia

Hair cell loss in the human inner ear leads to sensorineural hearing loss and vestibular dysfunction. Recent studies suggest that exogenous addition of growth factors, for example, transforming growth factor-α with insulin, may stimulate the production of new supporting cells and hair cells in the mature mammalian vestibular sensory epithelium. Before any growth factor can be seriously considered for the treatment of clinical problems related to hair cell loss, its effects on the extrasensory epithelia must also be fully explored. The aim of this study was to determine whether transforming growth factor-α and insulin stimulate cell proliferation in rodent vestibular extrasensory epithelia. The cell proliferation marker, tritiated thymidine, was infused along with transforming growth factor-α, insulin, or transforming growth factor-α plus insulin into the inner ears of adult rats via osmotic pumps. Effects of the test agents were assessed on normal and drug-damaged utricles. Drug damage was produced by delivering gentamicin directly into the inner ear before the infusion of test agent. Animals were killed 4 or 10 days after pump placement. Utricles were sectioned, processed for autoradiography, and examined for labeled cells within the extrasensory epithelia. In normal animals, transforming growth factor-α plus insulin stimulated DNA synthesis in all regions of the extrasensory epithelia, suggesting that these agents are mitogenic for these tissues. (Otolaryngol Head Neck Surg 1998;118:816-24.)     

Skeletal growth factor and other growth factors known to be present in bone matrix stimulate proliferation and protein synthesis in human bone cells

The purpose of the study was to investigate the effect of skeletal growth factor/insulinlike growth factor II and other growth factors known to be present in bone matrix on the proliferation and differentiation of human bone cells. Cells were isolated by collagenase digestion from femoral heads obtained during hip replacement operations. Cells were cultured in DMEM medium with 10% calf serum. Third to fifth passage cells were plated in multiwell plates and the medium changed to low serum (0.1%) for 2 days. The medium was changed to serum-free medium prior to addition of growth factors. Cell proliferation was measured by the incorporation of [3H]thymidine into DNA and by the percentage of cells that incorporate bromodeoxyuridine. Protein synthesis was measured by the incorporation of [3H]proline into trichloroacetic acid-precipitable material. Skeletal growth factor/insulinlike growth factor II and insulinlike growth factor I stimulated cell proliferation and protein synthesis in a dose-dependent manner. Alkaline phosphatase-specific activity was not increased by these factors. Transforming growth factor beta 1 did not affect cell proliferation but stimulated protein synthesis and increased the specific activity of alkaline phosphatase. Fibroblast growth factor did not affect any of the cell parameters. These studies suggest that skeletal growth factor/insulinlike growth factor II, insulinlike growth factor I, and transforming growth factor beta 1 may play a role in the local control of the proliferation and differentiation of human osteoblasts.     

RB1抑癌基因对人膀胱癌细胞抑癌作用的研究

采用电穿孔方法将组装于ＤＮＡ质粒载体ｐＣＭＶ－ｎｅｏ的ＲＢ１基因导入该基因功能丧失的膀胱癌细胞系ＨＴＢ９－５６３７。结果表明ＲＢ１抑癌基因对人类膀胱癌细胞具有明显抑癌作用。     

腺病毒载体介导的PML生长抑制因子对前列腺癌细胞生长和致瘤能力的抑制效果

为探讨用腺病毒载体携带ＰＭＬ（ＰｒｏｍｙｅｌｏｃｙｔｉｃＬｅｕｋｅｍｉａ）基因作为前列腺癌基因治疗的可能性，应用重组人携带ＰＭＬ基因腺病毒（ＡｄＰＭＬ）感染培养的前列腺癌细胞，观察表达ＰＭＬ蛋白的癌细胞与对照组癌细胞的体外生长和裸鼠体内致瘤能力变化，对荷瘤裸鼠瘤体周围注射ＡｄＰＭＬ，观察治疗组和对照组肿瘤生长的变化。结果显示，感染ＡｄＰＭＬ的前列腺癌细胞体外生长和裸鼠体内致瘤能力明显下降，荷瘤裸鼠瘤体周围注射ＡｄＰＭＬ后肿瘤生长速度明显减慢。证实了ＰＭＬ是一种生长抑制因子，提示其可能被应用于前列腺癌的基因治疗研究     

Adenovirus mediated gelsolin gene therapy for orthotopic human bladder cancer in nude mice

PURPOSE: Gelsolin is an actin regulatory protein that is undetectable or reduced in human bladder tumors compared with normal epithelial cells. Whether the over expression of gelsolin could inhibit tumor growth was investigated in an orthotopic bladder cancer nude mouse model using recombinant adenovirus encoding wild-type gelsolin (Ad-GSN). MATERIALS AND METHODS: The 2 human bladder cancer cell lines KU-7 and UMUC-2 were transduced with Ad-GSN in vitro. Flow cytometric analysis was done to examine the cell cycle after transducing the adenovirus. Cell growth was compared with control groups of these cells transduced with adenovirus containing the Escherichia coli beta-galactosidase gene Ad-betagal. In vivo KU-7 cells were introduced into the bladder of nude mice (day 0), followed by 3 injections into the urethra (days 2 to 4) with Ad-GSN or Ad-betagal (1 x 10 pfu). At 8 days after initial adenovirus exposure (day 10) each bladder was sectioned and stained, and the mass of the tumor was digitally determined. RESULTS: Bladder cancer cell growth (KU-7 and UMUC-2) was inhibited after these cells were transduced with Ad-GSN in vitro. Based on flow cytometric analysis over expression of gelsolin may cause these cells to arrest or delay at the G2/M phase of the cell cycle. In the orthotopic bladder cancer model the mass of the tumor was approximately 90% less in Ad-GSN treated animals than in controls. CONCLUSIONS: Ad-GSN provides a significant tumor suppressive effect on human bladder cancer cells in this orthotopic nude mouse model. Adenovirus mediated over expression of gelsolin may be useful therapy for human bladder cancer.