摘要
目的 观察串联质谱技术在乳腺癌血清蛋白表达中的差异。方法 选取2020年2月~2021年2月黑龙江省牡丹江医学院附属二院收治的乳腺癌新辅助化疗患者20例作为研究组,包含10例化疗有效患者及10例化疗无效患者,同时选取8名健康者作为对照组。采集所有患者的肘静脉血,分别采用高通量miRNA、iTRAQ表达谱测序技术,从“组学”的角度对乳腺癌新辅助化疗不同疗效患者miRNA差异表达谱、外周血单个核细胞蛋白组进行研究。随后,先进行Pathway通路分析,该过程在蛋白质层面进行,然后找出sRNA的成熟体序列,在此过程中严格根据miRbase18数据库。将已知的cDNA序列整理为一行,对得到的sRNA对应靶基因进行预测,Pathway分析靶基因,最后将同一Pathway通路中的sRNA靶基因及蛋白质寻找出来。结果 研究组和对照组共筛选出19个差异表达蛋白,选取其中具有较大表达差异的肝细胞生长因子(HGF)进行验证。iTRAQ标记图谱显示,研究组和对照组的血清HGF表达水平比较,差异有统计学意义(P<0.05)。研究组Ⅰ期、Ⅱ期、Ⅲ期患者的血清HGF表达水平逐渐升高(P<0.05),均高于对照组(均P<0.05),符合i TRAQ标记图谱鉴定结果。结论 串联质谱技术用于筛选发现乳腺癌细胞HGF表达水平升高具有显著效果,可有效指导临床诊治乳腺癌的工作,将有效参考提供给靶向治疗药物的开发。
Objective To analyze the effectiveness of isobaric tags for relative and absolute quantitation(iTRAQ)-based tandem mass spectrometry for systematic screening of serum protein expression differences in breast cancer patients. Methods A total of 20 patients with neoadjuvant chemotherapy for breast cancer were selected as the study group, which contained 10 patients with effective chemotherapy and 10 patients with ineffective chemotherapy, and 8healthy subjects were selected as the control group at the same time. Elbow venous blood was collected from all patients, and high-throughput sequencing in micro-ribonucleic acid(miRNA) expression profiling and iTRAQ technique were used to study the differential miRNA expression profiles and peripheral blood mononuclear cell proteome of patients with different efficacy of neoadjuvant chemotherapy for breast cancer from the perspective of “omics”. Subsequently, the Pathway analysis was performed at the protein level, and then the mature sequences of small RNA(sRNA)were identified, in which the miRbase18 database was strictly followed. The known complementary deoxyribonucleic acid(cDNA) sequences were organized into one line, the target genes corresponding to the obtained sRNA were predicted and analyzed by the Pathway analysis, and finally the s RNA target genes and proteins in the same Pathway were screened out, and the correlation between differential proteins and miRNA target genes was analyzed. Results A total of19 differentially expressed proteins in the study and control groups were screened out, and hepatocyte growth factor(HGF) with great expression differences was selected for validation. i TRAQ labeling-based profiles showed that there was a statistically significant difference between study group and the control group in serum HGF expression levels(P<0.05). The Western Blot validated and proved that serum HGF expression levels in patients of stage Ⅰ, stage Ⅱ and stage Ⅲ in the study group were progressively increased(P<0.05), and were higher than those in
作者
刘艳翠
张欣
李文媛
孙成
孙平
赵微
LIU Yancui;ZHANG Xin;LI Wenyuan;SUN Cheng;SUN Ping;ZHAO Wei(Teaching and Research Room of Anatomy,Mudanjiang Medical University of Heilongjiang Province,Mudanjiang 157011,China;Department of Ultrasound,the Second Affiliated Hospital of Mudanjiang Medical University of Heilongjiang Province,Mudanjiang 157011,China)
出处
《中国现代医生》
2022年第16期33-35,共3页
China Modern Doctor
基金
黑龙江省省属高等学校基本科研业务费科研项目(2020-KYYWFMY-0040
2020-KYYWFMY-0042)。
