CTNNA1基因启动子区异常甲基化与急性髓系白血病的相关性分析

目的研究CTNNA1基因启动子区异常甲基化与急性髓系白血病(AML)发生的相关性。方法选取180例AML患者骨髓标本及24例健康供者骨髓标本,通过甲基化特异性PCR方法(MS-PCR)进行CTNNA1基因启动子区异常甲基化阳性率进行检测,提取基因组DNA,设计硫化测序PCR(BS-PCR)引物及甲基化特异性PCR(MS-PCR)引物,进行PCR扩增及测序分析。结果 5例健康者标本及5例AML患者标本DNA经硫化且BS-PCR测序分析,其健康者标本甲基化率分别为1.5%、1.0%、1.0%、1.5%、1.0%,而AML患者CTNNA1甲基化率分别为92.0%、78.5%、86.0%、56.0%、90.0%,远远高于健康供者。MS-PCR分析结果显示,CTNNA1基因在24例健康人中呈完全非甲基化状态,在180例AML患者中其甲基化阳性率为37.2%(P<0.05)。结论 CTNNA1基因启动子区异常甲基化可能参与AML的发生,为疾病的早期监测提供分子理论依据。

miR-9-5P靶向CDH1、CTNNA1和ITGA6对脉络膜黑色素瘤细胞功能的影响

目的探讨miR-9-5P对脉络膜黑色素瘤细胞增殖、凋亡及迁移、侵袭的调控作用,并探寻其发挥作用的靶基因CDH1,CTNNA1和ITGA6。方法体外给予脉络膜黑色素瘤细胞转染阴性对照(NC)和miR-9-5p mimics。采用RTCA实时增殖实验检测细胞的增殖情况;EDU实验进一步检测细胞的增殖变化;流式细胞技术检测细胞的凋亡变化;Transwell实验观察细胞迁移和侵袭能力的变化情况;Targetscan生物信息学网站分析预测miR-9-5P的靶基因,并通过GO分类和KEGG富集分析找到与迁移和侵袭相关靶基因;Western blotting检测CDH1蛋白表达情况。结果RTCA实时增殖实验结果显示,miR-9-5p能够抑制脉络膜黑色素瘤细胞MUM-2B(t=6.925,P<0.05)及MUM-2C(t=4.762,P<0.05)的增殖;EDU实验结果显示,miR-9-5p能够抑制脉络膜黑色素瘤细胞MUM-2B(t=4.779,P<0.05)和MUM-2C(t=4.514,P<0.05)的增殖;流式细胞检测结果表明,miR-9-5p能够促进脉络膜黑色素瘤细胞MUM-2B(t=7.385,P<0.05)和MUM-2C(t=4.893,P<0.05)的凋亡;Transwell实验结果显示,miR-9-5p能够抑制细胞的迁移和侵袭;生物信息学预测及分析得出与细胞迁移侵袭相关的3个miR-9-5p的靶基因CDH1,CTNNA1和ITGA6;Western blotting证明CDH1是miR-9-5p的靶基因。结论miR-9-5p能抑制脉络膜黑色素瘤细胞的增殖、迁移和侵袭,促进脉络膜黑色素瘤细胞的凋亡,并通过调节CDH1,CTNNA1和ITGA6表达发挥调控作用。

急性髓系白血病CTNNA1基因的表达及临床意义

目的:研究急性髓系白血病(acute myeloid leukemia,AML)患者钙黏蛋白相关蛋白(α-catenin,CTNNA1)基因的表达情况并探讨其临床意义。方法:应用实时定量PCR(qRT-PCR)法检测25例非恶性血液病患者(对照组)和92例初诊AML患者(观察组)中CTNNA1基因的表达水平,并分析其与临床参数的相关性。结果:观察组中51例(51/92,55.43%)存在CTNNA1低表达,而对照组未发现CTNNA1低表达(0.00%),两组比较,差异有统计学意义(χ2=17.152,P<0.001);CTNNA1表达水平与AML患者的年龄、性别、外周血白细胞计数、血小板计数、血红蛋白含量、FAB分型、核型分组之间均无相关性(P均>0.05)。ROC曲线分析表明CTNNA1表达水平能有效鉴别观察组和对照组,AUC=0.806。观察组中CTNNA1低表达的患者化疗后完全缓解率高于对照组,但差异无统计学意义(χ2=2.476,P=0.142)。4例初诊AML患者在获得完全缓解后CTNNA1表达水平均较治疗前有所升高。结论:CTNNA1基因低表达可能是AML中的一个常见分子事件,检测其表达可用于AML患者的辅助诊断和疾病状态监测。

Hereditary diffuse gastric cancer: What the clinician should know

Hereditary diffuse gastric cancer(HDGC) is an inherited autosomal dominant syndrome with a penetrance of up to 80% affecting diverse geographic populations. While it has been shown to be caused mainly by germline alterations in the E-cadherin gene(CDH1), problematically, the genetic diagnosis remains unknown in up to 60% of patients. Given the important knowledge gaps regarding the syndrome, asymptomatic carriers of CDH1 mutations are advised for a prophylactic total gastrectomy. Intensive annual endoscopic surveillance is the alternative for carriers who decline gastrectomy. As HDGCs have a prolonged indolent phase, this provides a window of opportunity for surveillance and treatment. Recent findings of other gene defects in CTNNA1 and MAP3K6, as well as further characterization of CDH1 mutations and their pathogenicity will change the way HDGC patients are counselled for screening, surveillance and treatment. This review will bring the reader up to date with these changes and discuss future directions for research; namely more accurate risk stratification and surveillance methods to improve clinical care of HDGC patients.

Germline mutations in hereditary diffuse gastric cancer

Gastric cancer is one of the leading causes of cancer-related deaths worldwide. Among which, about 1%–3% of gastric cancer patients were characterized by inherited gastric cancer predisposition syndromes, knowing as hereditary diffuse gastric cancer（HDGC）. Studies reported that CDH1 germline mutations are the main cause of HDGC. With the help of rapid development of genetic testing technologies and data analysis tools, more and more researchers focus on seeking candidate susceptibility genes for hereditary cancer syndromes. In addition, National Comprehensive Cancer Network（NCCN） guidelines recommend that the patients of HDGC carrying CDH1 mutations should undergo prophylactic gastrectomy or routine endoscopic surveillances. Therefore, genetic counseling plays a key role in helping individuals with pathogenic mutations make appropriate risk management plans. Moreover, experienced and professional genetic counselors as well as a systematic multidisciplinary team（MDT） are also required to facilitate the development of genetic counseling and benefit pathogenic mutation carriers who are in need of regular and standardized risk management solutions. In this review, we provided an overview about the germline mutations of several genes identified in HDGC, suggesting that these genes may potentially act as susceptibility genes for this malignant cancer syndrome. Furthermore, we introduced information for prevention, diagnosis and risk management of HDGC. Investigations on key factors that may have effect on risk management decision-making and genetic data collection of more cancer syndrome family pedigrees are required for the development of HDGC therapeutic strategies.

急性髓系白血病中医分型与多项基因突变关系

目的探讨急性髓系白血病(AML)中医分型与CTNNA1基因启动子区异常甲基化、TET2、FLT3-ITD、CEBPA、DNMT3A、NPM1和TP53基因突变的关系。方法选择64例AML初诊患者,按中医证型诊断标准分为气阴两虚型、毒热炽盛型、瘀血痰结型。同时选择10名健康志愿者。收集患者年龄、性别、外周血情况、细胞遗传学危险分层、核型及预后等临床资料。采用甲基化特异性PCR方法检测CTNNA1基因启动子区异常甲基化,采用PCR扩增产物测序法检测TET2、FLT3-ITD、CEBPA、DNMT3A、NPM1和TP53基因突变情况。结果各证型性别、年龄、初诊外周血HGB、PLT、原始细胞比例及亚型比较,差异均无统计学意义(P>0.05)。各证型患者外周血WBC比较,差异有统计学意义(P<0.05)。与健康人比较,AML患者CTNNA1基因异常甲基化差异有统计学意义(P<0.05),但AML不同中医证型间CTNNA1基因异常甲基化差异无统计学意义(P>0.05)。AML不同中医证型间,相关突变基因数量、预后良好基因比例和预后危险分组情况等差异有统计学意义(P<0.05)。结论不同AML中医证型存在突变基因差异,相关突变基因可以辅助AML的中医辨证分型诊断。