Evaluation of DNA/RNA quality from cell block of malignant mesothelioma and lung adenocarcinoma.
Evaluation of DNA/RNA quality from cell block of malignant mesothelioma and lung adenocarcinoma.
复制标题
恶性间皮瘤和肺腺癌细胞块 DNA/RNA 质量评价。
DOI：
10.1002/dc.24960
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Ohbayashi C.
中科院分区：
文献类型：
--
作者：
Tatsumi S;Takeuchi M;Fujii T;Sugimoto A;Sekita-Hatakeyama Y;Nishikawa T;Uchiyama T;Morita K;Itami H;Takeda M;Yamazaki M;Ohbayashi C.
恶性间皮瘤(MM)是一种罕见的高度致命性肿瘤，起源于胸部和腹部表面的间皮组织。对包括胸水和腹水在内的体液进行细胞学检查，对于区分恶性间皮瘤和其他癌症，如肺癌、胃肠癌和转移性肿瘤是必不可少的。为了评估细胞块制备程序的有效性，该程序用于免疫细胞化学染色和肿瘤特异性基因突变的遗传小组分析，我们使用了各种固定剂。方法用恶性间皮瘤细胞系MESO4、H2 6和非小细胞肺癌细胞系HCC78制备细胞块。用10%中性缓冲福尔马林和四种不同的液体细胞学固定剂固定细胞。结果苏木精-伊红染色和免疫细胞学染色鉴定细胞块的形态，DNA/RNA抽提、定量聚合酶链式反应和下一代序列分析鉴定细胞块的核酸质量。D2-40和WT1染色因固定液和细胞团形成方法的不同而不同，但核酸降解程度不受任何方法的影响。结论尽管细胞学标本的形态评价受到细胞块制备方法的影响，但只要有足够的肿瘤细胞，D2-40和WT1染色仍可用于核酸提取和基因板分析。
Malignant mesothelioma (MM) is a rare and highly lethal tumor that arises from mesothelial tissue on the surface of the chest and abdominal cavity. Cytological examination of body fluids, including pleural fluid and ascites, is essential for the differentiation of malignant mesothelioma from other carcinomas, such as lung and gastrointestinal carcinomas and metastatic tumors. To evaluate the effectiveness of cell block preparation procedures, which are used for immunocytochemical staining and genetic panel analysis of tumor‐specific gene mutations, we used various fixatives. We also evaluated the effects of immunostaining, and the quality of nucleic acids for genetic analysis.MethodsCell blocks were prepared using the malignant mesothelioma cell lines MESO4 and H226 and non‐small cell lung carcinoma cell line HCC78. The cells were fixed using 10% neutral buffered formalin and four different fixatives for liquid cytology. Fixed cells were formed into cell clusters using sodium alginate or centrifugation, and paraffin‐embedded cell blocks were prepared.ResultsCell blocks were morphologically evaluated by hematoxylin and eosin and immunocytological staining, and the nucleic acid quality was evaluated by DNA/RNA extraction, qPCR, and next‐generation sequence analysis. D2‐40 and WT1 staining differed depending on the fixation solution and the cell cluster formation method; however, the degree of nucleic acid degradation was not impaired by any method.ConclusionAlthough the morphological evaluation of cytology specimens is affected by the method of cell block preparation, it is still useful for nucleic acid extraction and gene panel analysis, as long as there are sufficient amounts of tumor cells.