Kim Hyeon Woo, Jeong Dawoon, Ham Juyeon, Kim Heejoo, Ji Hwee Won, Choi Eun Ha, Kim Sun Jung
Department of Life Science, Dongguk University-Seoul, Goyang 10326, Republic of Korea.
Plasma Bioscience Research Center, Kwangwoon University, Seoul 01897, Republic of Korea.
Oxid Med Cell Longev. 2020 May 5;2020:9490567. doi: 10.1155/2020/9490567. eCollection 2020.
Cold atmospheric plasma (CAP) has been recognized as a potential alternative or supplementary cancer treatment tool, which is attributed by its selective antiproliferation effect on cancer cells over normal cells. Standardization of the CAP treatment in terms of biological outputs such as cell growth inhibition and gene expression change is essential for its clinical application. This study aims at identifying genes that show consistent expression profiles at a specific CAP condition, which could be used to monitor whether CAP is an appropriate treatment to biological targets. To do this, genes showing differential expression by two different CAP treatment conditions were screened in the MCF-7 breast cancer cells. As a result, ZNRD1 was identified as a potential marker with being consistently upregulated by 600 s but downregulated by the 10 × 30 s CAP treatment scheme. Expression of ZNRD1 was increased in breast cancer tissues compared to normal tissues, judged by cancer tissue database analysis, and supported by the antiproliferation after siRNA-induced downregulation in MCF-7. Interestingly, the antisense long noncoding RNA (lncRNA) of ZNRD1, ZNRD1-AS1, was regulated to the opposite direction of ZNRD1 by CAP. The siRNA-based qPCR analysis indicates that ZNRD1 downregulates ZNRD1-AS1, but not . ZNRD1-AS1 was shown to increase a few cis-genes such as HLA-A, HCG9, and PPP1R11 that were also regulated by CAP. Altogether, this study identified a pair of gene and its antisense lncRNA of which expression is precisely controlled by CAP in a dose-dependent manner. These genes could help elucidate the molecular mechanism how CAP regulates lncRNAs in cancer cells.
冷大气等离子体（CAP）已被公认为一种潜在的替代或辅助癌症治疗工具，这归因于其对癌细胞具有比正常细胞更强的选择性抗增殖作用。就细胞生长抑制和基因表达变化等生物学效应而言，CAP治疗的标准化对于其临床应用至关重要。本研究旨在鉴定在特定CAP条件下表现出一致表达谱的基因，这些基因可用于监测CAP对生物靶点是否为合适的治疗方法。为此，在MCF-7乳腺癌细胞中筛选了在两种不同CAP治疗条件下表现出差异表达的基因。结果，ZNRD1被鉴定为一个潜在标志物，在600秒的CAP处理下持续上调，但在10×30秒的CAP处理方案下下调。通过癌症组织数据库分析判断，与正常组织相比，ZNRD1在乳腺癌组织中的表达增加，并在MCF-7细胞中经siRNA诱导下调后出现抗增殖现象得到了证实。有趣的是，ZNRD1的反义长链非编码RNA（lncRNA），即ZNRD1-AS1，其表达受CAP调控的方向与ZNRD1相反。基于siRNA的qPCR分析表明，ZNRD1下调ZNRD1-AS1，但反之不然。结果显示ZNRD1-AS1增加了一些同样受CAP调控的顺式作用基因，如HLA-A、HCG9和PPP1R11。总之，本研究鉴定出一对基因及其反义lncRNA，它们的表达受CAP以剂量依赖方式精确调控。这些基因有助于阐明CAP在癌细胞中调控lncRNAs的分子机制。
