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研究发现结肠癌的新靶蛋白

By LabMedica International staff writers
Posted on 27 Aug 2018
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图片:Sensiscript逆转录试剂盒非常适合使用少量RNA的高灵敏度应用场合(图片蒙凯杰公司惠赐)。
图片:Sensiscript逆转录试剂盒非常适合使用少量RNA的高灵敏度应用场合(图片蒙凯杰公司惠赐)。

大肠癌(CRC)是美国诊断出来的第三大癌症暨美国男女性癌症致死的第三大病因。调控大肠癌形成过程的介导病原体是预测预后情况、疾病复发及治疗效果的理想的生物标志物,可改进大肠癌患者的诊疗。

 

大肠癌的治疗正变得越来越个性化,需要综合考虑个人的临床特征、肿瘤特征和分子特征。原癌基因β-连环蛋白会刺激大肠癌形成。卡西塔斯B种系淋巴瘤(c-Cbl)通过不为人知的机制靶向细胞核内的β-连环蛋白,从而抑制大肠肿瘤的生长。此外,c-Cbl对于人大肠癌的作用仍有许多未解之谜。

 

美国马萨诸塞州波士顿大学医学院(www.bumc.bu.edu)的科学家评估了一项单中心、回顾性观察队列研究,涉及72名四期大肠癌患者,他们有供分析用的肿瘤组织样本,在200411日至20141231日期间接受了治疗。科研小组获取了6微米厚的未染色石蜡包埋切片,这些切片已经过处理和检查。科学家专门开发了一种基于颜色的图像分割算法,估计人大肠癌样本表达c-Cbl的数量。

 

科学家的研究使用了蛋白质印迹、细胞培养、抗体、体外血管生成测定、泛素化测定、血管内皮生长因子(VEGF)酶联免疫吸附测定和成球试验。用德国希尔登凯杰公司(www.qiagen.com)RNeasy微型试剂盒提取HT-29细胞中表达各种不同构造的总RNA。研究人员用了凯杰公司的Sensiscript逆转录试剂盒,然后用美国加利福尼亚州福斯特市应用生物系统公司(www.appliedbiosystems.com)的预先确证过的人类AXIN2MYCVEGF引物及SYBRgreen进行RT-PCR

 

科研小组使用了新颖的定量组织病理技术证明,高度表达c-Cbl的肿瘤患者生存期中位数为3.7年,显著好于低度表达c-Cbl的肿瘤患者(1.8年),且存活三年的机会是后者的两倍多。他们还证明,c-Cbl调控细胞核内的β-连环蛋白需要c-Cbl Tyr371的磷酸化,因为其突变削弱了它靶向β-连环蛋白的能力。酪氨酸371 (Y371H)变体与细胞核β-连环蛋白相互作用,却不能使之泛素化。c-CblY371H变体局限在细胞核内,致使其对细胞核β-连环蛋白产生巨大的负面效应。

 

该研究的论文发表于2018717日的《美国病理学杂志》(American Journal of Pathology)。作者总结说,肿瘤的c-Cbl水平与大肠癌转移患者的总生存期有关联,进一步证实c-Cbl是大肠肿瘤生长的负向调控因子。这些成果表明Wnt/β-连环蛋白的信号可能在下游调控CBLc.1111T>C (p.371Y>H)处突变的影响,并确证了c-Cbl的分子特征,这些特征对于它如何影响细胞核β-连环蛋白至关重要。

Related Links:

波士顿大学医学院>>> www.bumc.bu.edu

凯杰 >>> www.qiagen.com

应用生物系统公司>>> www.appliedbiosystems.com



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