甲磺酸伊马替尼调控Telocytes对下颌下腺唾液分泌功能的影响

目的探讨甲磺酸伊马替尼调控Telocytes(TCs)对下颌下腺唾液分泌功能的影响。方法采用免疫荧光染色技术对TCs免疫标记物(C-kit/CD117、CD34)进行双标记染色,进而对小鼠下颌下腺内TCs进行定位。采用透射电子显微镜(TEM)显示小鼠下颌下腺内TCs的超微结构及其与周围细胞之间的关系。建立甲磺酸伊马替尼干预模型,实验小鼠分5组:正常组(共24只),用药1周、2周、3周、4周组(共24只),药物以80 mg/(kg·d)每日灌胃,免疫荧光显示,用药前后下颌下腺内TCs的变化;免疫印迹法(Western blotting)观察用药前后CD117、CD34及下颌下腺内α唾液淀粉酶(α-Amy)蛋白表达水平的变化。结果免疫荧光染色显示,TCs广泛分布于下颌下腺结缔组织内,胞体较小有突起(Tps),Tps相互连接成网络样结构,包绕着腺泡及导管,随甲磺酸伊马替尼干预时间延长,用药组Tps构成的网络样结构变稀疏。超微结构显示,Tps呈念珠状,与邻近组织紧密相连,周围可见胞外囊泡。随用药时间的增加Tps减少,TCs胞内囊泡增多,胞内细胞器减少。免疫印迹法显示,用药后CD117、CD34、α-Amy蛋白表达水平相应减少且互为正相关。结论小鼠下颌下腺内存在TCs,甲磺酸伊马替尼对TCs的干预可能会通过影响TCs的结构、免疫表型和细胞间通讯降低下颌下腺唾液分泌功能。     

小肠的恶性胃肠道间质瘤伴淋巴结转移病理分析

目的 探讨伴淋巴结转移的小肠恶性胃肠道间质瘤的临床病理特征、c-kit基因突变情况,以及对甲磺酸伊马替尼(imatinib mesylate,Glivec)的治疗反应.方法 对2例发生在小肠伴发淋巴结转移的胃肠道间质瘤进行光镜观察、免疫组织化学标志及基因突变分析并随访甲磺酸伊马替尼的治疗效果.结果 2例均为小肠浆膜面肿块,镜下观察肿瘤均以梭形细胞为主,伴有少量上皮样细胞,呈多结节状,并出现大片凝固性坏死;免疫组织化学标志肿瘤细胞CD117阳性,基因突变检测发现均存在c-kit基因第11号外显子的突变.例1显示第11号外显子559～569位点杂合性缺失,伴570、571位点TACATA杂合性突变为GACAGA;例2显示第11号外显子559～565杂合性缺失.结论 小肠胃肠道间质瘤伴淋巴结转移是一种少见病变,需要同发生在此处的其他恶性软组织肿瘤鉴别;该肿瘤对甲磺酸伊马替尼的治疗效果取决于c-kit基因的具体突变类型.     

小肠的恶性胃肠道间质瘤伴淋巴结转移病理分析

目的 探讨伴淋巴结转移的小肠恶性胃肠道间质瘤的临床病理特征、c-kit基因突变情况,以及对甲磺酸伊马替尼(imatinib mesylate,Glivec)的治疗反应.方法 对2例发生在小肠伴发淋巴结转移的胃肠道间质瘤进行光镜观察、免疫组织化学标志及基因突变分析并随访甲磺酸伊马替尼的治疗效果.结果 2例均为小肠浆膜面肿块,镜下观察肿瘤均以梭形细胞为主,伴有少量上皮样细胞,呈多结节状,并出现大片凝固性坏死;免疫组织化学标志肿瘤细胞CD117阳性,基因突变检测发现均存在c-kit基因第11号外显子的突变.例1显示第11号外显子559～569位点杂合性缺失,伴570、571位点TACATA杂合性突变为GACAGA;例2显示第11号外显子559～565杂合性缺失.结论 小肠胃肠道间质瘤伴淋巴结转移是一种少见病变,需要同发生在此处的其他恶性软组织肿瘤鉴别;该肿瘤对甲磺酸伊马替尼的治疗效果取决于c-kit基因的具体突变类型.     

胃肠道间质瘤患者伊马替尼血药浓度的监测及意义

胃肠间质瘤（GIST）是消化道常见的间叶源性肿瘤,多发生于胃部,其免疫组化CD117及DOG-1常表达阳性,介导Kit基因突变是导致该病的主要机制。伊马替尼是一种酪氨酸酶抑制剂,可以很好的抑制c-Kit的活性,作为复发、不可切除或者晚期的GIGT患者一线用药甲磺酸伊马替尼,它已被证实是治疗中晚期GIST患者最有效的药物,可以显著改善预后。在常规的治疗过程中,伊马替尼的血药浓度在不同的GIST患者中存在很大差异,既有血药浓度不足导致疗效较差,也有血药浓度过高导致不良反应太大,血药浓度的监测对于提高患者的依从性及预后具有重要的意义。同时我们也可以通过马替尼血药浓度的检测去探究疾病的原因,然而对于甲磺酸伊马替尼血药浓度的监测主要集中在国外,国内相关研究甚少。本文从伊马替尼的药代动力学、伊马替尼血药浓度的现状及问题、监测的意义等方面进行综述,探讨伊马替尼血药浓度的监测对GIST患者的意义。     

中国胃肠道间质瘤诊断治疗共识

胃肠道间质瘤(gastrointestinal stromal tumors,GIST)是胃肠道最常见的间叶源性肿瘤,免疫组织化学染色大多数呈CD117阳性.传统放疗和化疗对GIST几乎无效.酪氨酸激酶抑制剂甲磺酸伊马替尼的临床应用使GIST的治疗发生了重大的改变,为部分晚期GIST患者带来了延长生存期的疗效.同时由于对GIST基因突变认识的进一步提高,使GIST的诊断率显著提高.     

酪氨酸激酶抑制剂甲磺酸伊马替尼对PTEN介导的K562细胞侵袭的影响

目的:研究酪氨酸激酶抑制剂甲磺酸伊马替尼对K562细胞PTEN信号转导的调控,以及对细胞侵袭功能的影响.方法:不同浓度甲磺酸伊马替尼作用K562细胞不同时间后,通过荧光定量PCR检测BCR/ABL、PTEN、FAK水平变化及相互关系,免疫细胞化学染色检测FAK蛋白水平,Transwell小室检测K562细胞侵袭功能.结果:2μg/mL甲磺酸伊马替尼作用K562细胞在36 h内,随着BCR/ABL融合基因表达减低,PTEN mRNA表达上调,FAK mRNA及蛋白表达下调,K562细胞侵袭功能明显减弱.作用48 h后,随着BCR/ABL融合基因的抑制减弱,PrEN表达进而减低,而FAK表达升高.BCR/ABL mRNA与PTEN mRNA呈负相关趋势,与FAK mRNA呈正相关趋势.结论:甲磺酸伊马替尼通过抑制BCR/ABL融合基因调控PTEN/FAK信号转导通路,参与抑制白血病K562细胞侵袭作用.     

212例胃肠间质瘤的临床病理特征及预后分析

目的探讨胃肠间质瘤（GIST）的临床特征和分子病理学特点,分析影响GIST预后的相关因素。方法回顾性分析2004年4月至2011年8月南方医院收治的212例GIST患者的临床病理和随访资料,应用生存分析比较不同因素对预后的影响。对接受甲磺酸伊马替尼治疗的53例患者,采用基质辅助激光解析/电离-飞行时间质谱方法检测KIT和PDGFRa基因相关位点的突变情况。结果单因素生存分析显示肿瘤大小、核分裂数、美国国立卫生研究所（NIH）危险度分级、转移、手术及甲磺酸伊马替尼影响GIST患者的生存率。多因素生存分析提示,NIH危险度分级和甲磺酸伊马替尼是影响预后的独立因素。53例GIST患者中,KIT基因突变39例（73.6%）,其中外显子11突变21例（53.8%）,外显子9突变13例（33.3%）。KIT外显子11突变形式主要为5’端第557-558密码子缺失最常见;外显子9突变均为插入串联重复。未检测到PDGFRa基因突变的病例。结论 NIH危险度分级和甲磺酸伊马替尼治疗与GIST患者的生存密切相关,基因突变检测对指导生物靶向治疗和预测其疗效具有重要意义。     

吉非替尼通过增加Foxo3a活性抑制肺纤维化小鼠上皮-间质转分化

 目的：研究吉非替尼对肺纤维化小鼠转录因子叉头框蛋白O3a(Foxo3a)活性、α-平滑肌肌动蛋白（α-SMA）水平及相关通路的影响,探讨吉非替尼抑制肺上皮-间质转分化的可能机制。方法：将30只SPF级雌性昆明小鼠随机分为3组：对照组（生理盐水气管内雾化）、博来霉素组（博来霉素3 mg/kg溶于100 μL生理盐水气管内雾化）和吉非替尼处理组（博来霉素气管内雾化后,每天吉非替尼20 mg/kg溶于100 μL生理盐水灌胃）。实验第14天收集样本,将小鼠肺组织置于10%中性甲醛固定,石蜡包埋切片后行HE与Masson染色;RT-PCR法检测Foxo3a和α-SMA mRNA表达水平;Western blotting法检测表皮生长因子受体（EGFR）、Akt、Foxo3a和α-SMA蛋白表达水平。结果：吉非替尼处理组小鼠肺组织病理损伤较博来霉素组明显减轻,胶原沉积明显减少,炎症损伤评分及纤维化评分明显下降（均P<0.01）,Foxo3a mRNA表达水平明显升高（P<0.05）,α-SMA mRNA表达水平明显下降（P<0.05）,总Foxo3a蛋白表达增加,但Foxo3a磷酸化水平显著下降(P<0.01),胞核Foxo3a蛋白明显增加（P<0.05）;同时,EGFR和Akt磷酸化水平也显著下降（P<0.01,P<0.05）,上皮-间质转分化标志蛋白α-SMA表达水平明显降低（P<0.05）。结论:吉非替尼抑制博来霉素诱导的肺纤维化,其机制可能与抑制EGFR/Akt通路活化、增强转录因子Foxo3a活性、从而抑制上皮-间质转分化密切相关。     

小鼠下颌下腺内ncNOS的表达

目的对小鼠下颌下腺内ncNOS进行定性和定位研究。方法取8只昆明种小鼠下颌下腺,进行HE染色和免疫组织化学染色。结果小鼠下颌下腺纹状管及小叶间导管上皮细胞呈ncNOS免疫反应阳性,而腺泡细胞则为阳性。结论小鼠下颌下腺导管上皮中有ncNOS的表达,提示下颌下腺具有复杂的分泌功能。     

新生大鼠心telocytes的形态学和蛋白标记

目的探讨新生大鼠心telocytes(TCs)的形态学特征并验证其是否表达神经细胞、神经胶质细胞的特异性标记。方法酶消化法培养新生大鼠原代细胞,免疫细胞化学、扫描电子显微镜鉴定TCs,免疫荧光技术检测TCs的神经细胞和神经胶质细胞的蛋白表达。新生大鼠大脑组织免疫组织化学染色作为阳性对照。结果免疫组织化学检测显示波形蛋白(vimentin)表达阳性,免疫荧光检测vimentin和CD34共表达,但TCs内神经元特异性烯醇化酶(NSE)、GFAP、OX42、2’3’-环腺苷酸-3’-磷酸二酯酶(CNpase)表达阴性。扫描电子显微镜下TCs形态清晰典型,特征形态明显。结论大鼠心TCs虽然具有与神经细胞类似的形态,但其并不具有神经细胞的特性。     

Changing phenotype of gastrointestinal stromal tumours under imatinib mesylate treatment: a potential diagnostic pitfall

AIMS: The diagnosis of gastrointesinal stromal tumours (GISTs) is widely based on morphological features and KIT (CD117) immunoreactivity. Most patients with advanced GISTs show a major clinical response after treatment with imatinib mesylate. The histopathological features of GISTs in patients on prolonged imatinib treatment have, thus far, not been addressed in detail. In this report, we present three patients with metastatic GISTs, who received more than 1 year of therapy with imatinib, and whose tumours changed their morphological and immunohistochemical characteristics during continued treatment with the drug. METHODS AND RESULTS: All three primary GISTs from these patients were classical spindle-type tumours, showing diffuse, strong CD117, CD34, and focal alpha-smooth muscle actin expression. During treatment, two clinically progressive and one clinically stable GIST revealed a diffuse epithelioid, or pseudopapillary epithelioid growth pattern, characterized by rounded cells with eosinophilic cytoplasm and uniform round-to-ovoid nuclei. In addition, GIST specimens from patients on therapy showed complete loss of CD117 immunoreactivity. Remarkably, two of these tumours also became CD34 immunonegative and in one case the progression was accompanied by desmin expression. KIT mutational analysis revealed the presence of distinct exon 11 mutant isoforms in all cases examined, while the same genotype was sustained in the base line and on-therapy tumour specimens, proving the common origin of analysed specimens. CONCLUSIONS: GISTs subject to imatinib treatment can undergo striking (immuno)phenotypic changes, which are not necessarily corroborated by new genotypic modifications. Because these may mimic other tumour types, this feature creates a differential diagnostic challenge, of which the pathologist should be aware.     

异丙肾上腺素影响下颌下腺干/祖细胞增殖的数量还是能力？

背景：异丙肾上腺素注射能够促进啮齿类动物涎腺腺泡细胞增殖、肥大。然而,异丙肾上腺素注射后涎腺干/祖细胞的增殖能力目前仍不清楚。 目的：研究异丙肾上腺素腹腔注射对SD大鼠下颌下腺干/祖细胞激活、增殖能力。 方法：SD大鼠随机分为2组,分别腹腔注射异丙肾上腺素与生理盐水,连续注射5 d后取下颌下腺,酶消化法制取下颌下腺细胞悬液,体外培养,获得下颌下腺类上皮细胞集落并计数。挑取增殖较快的集落,进行CD90.1、层粘连蛋白、α6β1等免疫细胞化学检测。 结果与结论：与对照组相比,异丙肾上腺素注射组中、低增殖集落数量较少,高增殖集落数量较多,差异有显著性意义(P < 0.05),而集落总数无明显区别(P > 0.05)。高增殖下颌下腺类上皮细胞集落CD90.1、层粘连蛋白、α6β1抗体均为阳性表达。实验结果显示,异丙肾上腺素注射不能明显增加下颌下腺中的干/祖细胞数量,但是能提高腺体中干/祖细胞的增殖能力。     

BCR/ABL amplification in chronic myelocytic leukemia blast crisis following imatinib mesylate administration

The onset of accelerated phase or blast crisis of chronic myelocytic leukemia (CML) is usually associated with the acquisition of new chromosome abnormalities in addition to the t(9;22)(q34;q11) that is characteristic of the chronic phase CML. We describe the cytogenetic and molecular genetic findings in two cases of myelocytic blast crisis of CML, one occurring 6 months after commencing treatment with the ABL-specific tyrosine kinase inhibitor imatinib mesylate (STI571, Glivec, or Gleevec) and the second treated with imatinib mesylate for established blast crisis. In both cases, multiple secondary cytogenetic abnormalities were observed at transformation, with homogeneously staining regions that were shown to contain BCR/ABL amplification by fluorescence in situ hybridization appearing after imatinib mesylate administration. BCR/ABL amplification is emerging as an important mechanism of acquired resistance to imatinib mesylate.     

Epithelioid gastrointestinal stromal tumor with PDGFRA activating mutation and immunoreactivity.

The authors report a unique case of an intra-abdominal, epithelioid mesenchymal tumor that had an activating mutation of PDGFRA and a strong PDGFRA immunoreactivity but lacked both c-kit mutation and c-kit protein (CD117) expression. IHC study showed that the tumor cells were diffusely and strongly positive for PDGFRA, vimentin, CD34, and Bcl-2 but completely negative for CD117 as well as for muscle, epithelial, endothelial, endocrine, mesothelial, neural, and melanocytic cell markers. Molecular study revealed a mutation at the juxtamembrane domain of exon 12 in PDGFRA gene with GTC to GAC transition at codon 561 (V561D), as shown in the previous mutational studies on gastrointestinal stromal tumor (GIST). This case likely represents an example of GIST with PDGFRA activating mutation and PDGFRA immunoreactivity without CD117 positivity, which has not been documented in the literature. STI 571 (imatinib mesylate [Gleevec]) might be an effective therapy in this case, since Gleevec targets both PDGFRA and c-kit oncoproteins.     

Dedifferentiated gastrointestinal stromal tumor arising de novo from the small intestine

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract and usually display monotonous cytologic features and immunoactivity for CD117. Anaplastic GIST, with pleomorphic cells and loss of CD117, until recently have only been reported in patients with chronic imatinib mesylate treatment. Dedifferentiated GISTs arising de novo is a newly identified entity that may prove to be difficult to diagnose. We present the case of a 52-year-old female found to have a dedifferentiated GIST without prior imatinib mesylate therapy. This case is the first reported dedifferentiated GIST arising de novo from the small bowel, and at 30 cm in greatest diameter, the largest reported to date. Additionally, we demonstrate for the first time the loss of DOG1 in the anaplastic component of the tumor. De novo dedifferentiated GIST is a rare and diagnostically challenging tumor that may be mischaracterized unless considered in the differential diagnosis.     

Ligamentum arteriosum and its telocytes: An ultrastructure description

Results from my study have shown in 2022 the existence of telocytes (TCs) in mice ligamentum arteriosum (LA). Telocytes (TCs) are unique interstitial or stromal cells of mesodermal origin, defined by long cellular extensions called telopodes (Tps) which form a network, connecting them to surrounding cells. These Tps have dilated portions named podoms (usually containing mitochondria, endoplasmic reticulum and caveolae) and very thin segments (below resolving power of light microscopy), called podomers. Generally, transmission electron microscope revealed the existence of Tps with various conformations: (a) long, flattened irregular veils (ribbon-like segments) with knobs, corresponding to podoms, and (b) tubular structures (podomers) with uneven caliber because of irregular dilations (knobs)—the podoms. Also shown were numerous extracellular vesicles and exosomes released by the TCs which sometimes made direct contact with telopodes. Telopodes were observed communicating with each other through adherens junctions. Telopodes sandwiched between myocytes or in close proximity (0.01 μm) from nerve terminals were also observed. These data might be useful for understanding the role(s) of TCs in intercellular signaling and communication, neuromodulation as well as comprehension of pathologies like structural remodeling within the LA.     

Advanced gastrointestinal stromal tumors successfully treated with imatinib mesylate: a report of two cases

Activation of kit-receptor tyrosine kinase occurs in all cases of gastrointestinal stromal tumors, regardless of the mutation status of kit. Imatinib mesylate (STI 571,Gleevec) is a selective inhibitor of certain protein tyrosine kinases. It has been shown in preclinical models and clinical studies to have activity against such tumors. The aim of the present study was to report the efficacy of imatinib mesylate in the treatment of advanced gastrointestinal stromal tumors. Two adults with histologically confirmed, unresectable, and metastatic gastrointestinal stromal tumors that expressed CD117 (a marker of kit-receptor tyrosine kinase) were identified at our institution during 2000-2002. As the diseases were advanced and not amenable to surgery, chemotherapy, or radiation therapy, imatinib mesylate was used, because this targeted inhibitor has been shown to be active against advanced gastrointestinal stromal tumors and has a mild toxicity profile. Imatinib mesylate induced a sustained response in both patients with advanced unresectable or metastatic gastrointestinal stromal tumors. Inhibition of the KIT signal-transduction pathway is a promising treatment for advanced gastrointestinal stromal tumors, which resist conventional chemotherapy.     

A lethal mesenteric gastrointestinal stromal tumor: a case report and review of the literature

Gastrointestinal stromal tumors (GISTs) arising from the mesentery are very rare. Here, we report a 53-year old man with a huge lobulated cystic-solid tumor in the left lower quadrant of the abdomen, which had been proved clinically and radiographically. Surgical resection showed that the large mass was noted at the mesentery of small intestine. Grossly, the largest diameter of the mass were measured up to 23 cm, and poorly circumscribed. Histological observation demonstrated it as a malignant GIST with positive CD117 (c-kit) staining. Mitotic figures were frequently observed up to 110 per 50 high power fields. Soon after the surgery, the patient experienced local recurrence with quick growth. The patient received targeted therapy (imatinib mesylate) but had no ideal effect. The patient died nine months after the operation because of rapid disease progression.     

Sequence analysis and high-throughput immunhistochemical profiling of KIT (CD 117) expression in uveal melanoma using tissue microarrays

We aimed to immunohistochemically examine the expression of KIT (CD 117) in human posterior uveal melanoma and to analyze KIT-positive tumors for gene mutations. Brought into a tissue microarray (TMA) format were 101 formalin-fixed, paraffin-embedded posterior uveal melanomas. Immunhistochemistry was performed using the polyclonal anti-CD117 antibody from Dako (A4502). In ten selected KIT-positive tumors, exons 2, 8, 9, 11, 13 and 17 were sequenced. Of the 101 cases, 89 (88%) could be evaluated on the TMAs. Immunohistochemistry for CD 117 was weakly positive in 5 cases (6%), moderately positive in 10 cases (12%) and strongly positive in 57 cases (69%). No KIT mutations were detected in the analyzed exons. In conclusion, human posterior uveal melanoma frequently expresses CD117 at high levels. Although KIT mutations could not be found, it appears justified to investigate the utility of imatinib mesylate in the treatment of these patients.