Objective To explore circulating biomarkers for screening the invasiveness of non-functioning pituitary adenomas(NF-PAs).Methods The exosomal RNAs were extracted from serum of patients with invasive NF-PA(INF-PA)or no...
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Objective To explore circulating biomarkers for screening the invasiveness of non-functioning pituitary adenomas(NF-PAs).Methods The exosomal RNAs were extracted from serum of patients with invasive NF-PA(INF-PA)or noninvasive NF-PA(NNF-PA).Droplet digital PCR was adapted to detect the mRNA expression of candidate genes related to tumor progression or invasion,such as cyclin dependent kinase 6(CDK6),ras homolog family member U(RHOU),and spire type actin nucleation factor 2(SPIRE2).Student’s t-test was used to analyze the statistical difference in the mRNA expression of candidate genes between the two *** operating characteristic(ROC)curve was used to establish a model for predicting the invasiveness of *** accuracy,sensitivity,specificity and precision of the model were then obtained to evaluate the diagnostic *** CDK6(0.2600±0.0912 vs.0.1789±0.0628,t=3.431,P=0.0013)and RHOU mRNA expressions(0.2696±0.1118 vs.0.1788±0.0857,t=2.946,P=0.0052)were upregulated in INF-PAs patients’serum exosomes as compared to *** CDK6,the area under the ROC curve(AUC)was 0.772(95%CI:0.600-0.943,P=0.005),the accuracy,sensitivity,specificity and precision were 77.27%,83.33%,75.00%and 55.56%to predict the invasiveness of *** RHOU,the AUC was 0.757(95%CI:0.599-0.915,P=0.007),the accuracy,sensitivity,specificity and precision were 72.73%,83.33%,68.75%and 50.00%.In addition,the mRNA levels of CDK6 and RHOU in serum exosomes were significantly positively correlated(r=0.935,P<0.001).After combination of the cut-off scores of the two genes,the accuracy,sensitivity,specificity and precision were 81.82%,83.33%,81.25%and 62.50%.Conclusions CDK6 and RHOU mRNA in serum exosomes can be used as markers for predicting invasiveness of *** of the two genes performs better in distinguishing INF-PAs from *** results indicate CDK6 and RHOU play important roles in the invasiveness of NF-PAs,and the established diagnostic method is valua
目的 应用Affymetrix全基因组芯片结合荧光定量PCR（quantitative real-timePCR，qPCR）技术，进行致病性DNA拷贝数变异的精细定位研究。方法以一个定位于染色体7q36的中国人遗传性三节拇指多并指综合征伴随Ⅳ型并指家系中的一例患者为研究对象。收集外周血标本，常规提取基因组DNA。应用Affymetrix Genome-Wide Human SNP Array6．0芯片，将基因组DNA纯化，经过酶切、连接、扩增、标记、杂交、染色和扫描等步骤后得到原始数据，应用Affymetrix Genotyping Console3．0软件进行拷贝数分析。在经芯片分析所确定的重复范围内设计引物，采用qPCR方法进行验证，并进一步缩小断端范周、精确重复区域范围。结果将患者重复区域两断端范围由原来的113kb和33kb分别缩小到5．4kb和1．8kb，致病性DNA重复范围由原来的291～437kb精确至379～387kb。结论应用Affymetrix全基因组芯片联合qPCR技术可以实现对DNA拷贝数突变的精确、可靠的检测。
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