实用医学杂志 ›› 2021, Vol. 37 ›› Issue (18): 2326-2331.doi: 10.3969/j.issn.1006⁃5725.2021.18.005

• 基础研究 • 上一篇    下一篇

PRKAA2基因在肿瘤细胞与血细胞中表达差异研究

孙春霞 ,陈书坤, 郑磊   

  1. 南方医科大学南方医院检验科(广州 510515)

  • 出版日期:2021-09-25 发布日期:2021-09-25
  • 通讯作者: 郑磊 E⁃mail:nfyyzhenglei@smu.edu.cn
  • 基金资助:
    国家杰出青年科学基金(编号:82025024);国家自然科学基金(编号:81902163);国家博士后科学基金(编号:2019M652972)

Differential expression of PRKAA2 gene in multiple tumor cell lines versus WBC

SUN Chunxia,CHEN Shukun,ZHENG Lei.   

  1. Department of Laboratory Medicine,Nanfang Hospital,Southern Medical University,Guang⁃ zhou 510515,China

  • Online:2021-09-25 Published:2021-09-25
  • Contact: ZHENG Lei E⁃mail:nfyyzhenglei@smu.edu.cn

摘要:

目的 研究常见肿瘤细胞与白细胞(WBC)中的脂代谢相关基因 PRKAA2 的表达差异,为 PRKAA2 用于循环肿瘤细胞(circulating tumor cells,CTCs)的鉴定提供理论基础。方法 根据 THPA(The human protein atlas)数据库的 RNA 测序数据,分析 WBC PRKAA2 基因表达丰度,应用实时荧光定量聚合 酶链反应(qRT⁃PCR)比较多种肿瘤细胞与 WBC PRKAA2 基因的差异表达。采用原位锁式探针杂交技术在细胞原位可视化检测 HepG2、U87、Hela、PC⁃3 和健康人外周血 WBC 的分子表达。最后,掺入实验模 拟临床真实样本验证 PRKAA2 用于鉴定肿瘤细胞的准确性。结果 与健康人外周血 WBC 比较,HepG2 U87、Hela PC⁃3 PRAKK2 的表达显著上调,差异有统计学意义(均 P < 0.05)。原位锁式探针杂交实验 结果显示 HepG2、U87、Hela、PC⁃3(均为 PRKAA2+)和 WBC(PRKAA2-)的分子表达有显著区别。掺入实验表明,利用 PRKAA2 mRNA 荧光信号能准确识别肿瘤细胞。结论 PRKAA2 HepG2、U87、Hela、PC⁃3 瘤细胞系中的表达量显著高于 WBC,且通过检测 PRKAA2 mRNA 的荧光信号可从大量的 WBC 背景细胞中准确识别肿瘤细胞。

关键词:

循环肿瘤细胞, 脂质代谢, 生物标志物, PRKAA2

Abstract:

Objective To study the differential expression of the lipid metabolism⁃related gene PRKAA2 in several common tumor cells and white blood cells(WBC),in order to provide a theoretical basis for the identifi⁃ cation of circulating tumor cells(CTCs)using PRKAA2. Methods RNA⁃sequencing data from The Human Protein Atlas(THPA)were used to infer the expression abundance of PRKAA2 in WBCs. The expression of PRKAA2 in multiple cancer cells and WBCs were evaluated with real ⁃time fluorescent quantitative polymerase chain reaction (qRT⁃PCR). In situ padlock probe hybridization technology was used to visually detect the PRKAA2 expression of HepG2,U87,Hela,PC⁃3 and healthy individuals WBCs. We finally spiked tumor cells in WBCs to mimic the clinical samples to verify the accuracy of PRKAA2 used to identify tumor cells. Results Compared with WBC of healthy individuals,the expression of PRAKK2 in HepG2,U87,Hela,and PC⁃3 cells was significantly up⁃regulated and the difference was statistically significant(all P < 0.05). In situ padlock probe hybridization experiments showed a clear distinction of the molecular expression between HepG2,U87,Hela,PC⁃3 cells(all PRKAA2+)and WBC(PRKAA2⁃). The PRKAA2 signal and CFSE fluorescence co⁃localized supporting the accuracy of PRKAA2 for the detection of tumor cells from Peripheral blood,which was confirmed using confocal microscopy. Conclusions The expression level of PRKAA2 in tumor cell lines HepG2,U87,Hela,PC⁃3 was significantly higher than that of WBC,and the difference in the fluorescence signal of PRKAA2 mRNA can be used to accurately identify tumor cells from a large number of WBCs.

Key words:

circulating tumor cells, lipid metabolism, biomarker, PRKAA2