achaete-scute complex homologue 1 (ASCL1) 在肺神经内分泌癌中的意义;使用小细胞肺癌细胞和 Ascl1 诱导的肺神经内分泌癌细胞进行 RNA 序列分析。

Significance of achaete-scute complex homologue 1 (ASCL1) in pulmonary neuroendocrine carcinomas; RNA sequence analyses using small cell lung cancer cells and Ascl1-induced pulmonary neuroendocrine carcinoma cells.

机构信息

Department of Pathology and Experimental Medicine, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Chuo-ku, Kumamoto, 860-8556, Japan.

Department of Pathology, Kumamoto Health Science University, Kumamoto, Japan.

出版信息

Histochem Cell Biol. 2020 Jun;153(6):443-456. doi: 10.1007/s00418-020-01863-z. Epub 2020 Mar 13.

Abstract

ASCL1 is one of the master transcription factors of small cell lung carcinoma (SCLC). To investigate the significance of ASCL1 in pulmonary neuroendocrine carcinoma, we performed 2 comparative RNA-seq studies between H69 (ASCL1-positive, classical type SCLC) and H69AR (ASCL1-negative, variant type SCLC) and between ASCL1-transfected A549 adenocarcinoma cell lines (A549(ASCL1+) cell lines) and A549(control) cell lines. RNA-seq analyses revealed that 940 genes were significantly different between the H69 and H69AR cell lines, and 728 between the A549(ASCL1+) and A549(control) cell lines. In total, 120 common genes between these analyses were selected as candidate ASCL1-related genes, and included genes with various cellular functions, such as neural development, secretion, growth, and morphology. Their expression degrees in three classical and two variant SCLC cell lines, two A549(ASCL1+) and two A549(control) cell lines were subjected to quantitative PCR analyses. Since the candidate ASCL1-related genes were strongly expressed in the classical SCLC and A549(ASCL1+) cell lines and more weakly expressed in the variant SCLC and A549(control) cell lines, the ASCL1-related 7 molecules INSM1, ISL1, SYT4, KCTD16, SEZ6, MS4A8, and COBL were further selected. These molecules suggested diverse functions for A549(ASCL1+): INSM1 and ISL1 are transcription factors associated with neuroendocrine differentiation, while SYT4, KTCD16, and SEZ6 may be related to neurosecretory functions and MS4A8 and COBL to cell growth and morphology. An immunohistochemistry of these seven molecules was performed on lung carcinoma tissues and the xenotransplanted tumors of A549(ASCL1+), and they were preferentially and positively stained in ASCL1-postive tumor tissues.

摘要

ASCL1 是小细胞肺癌(SCLC)的主要转录因子之一。为了研究 ASCL1 在肺神经内分泌癌中的意义,我们进行了 2 项比较 RNA-seq 研究,分别在 H69(ASCL1 阳性,经典型 SCLC)和 H69AR(ASCL1 阴性,变异型 SCLC)以及 ASCL1 转染的 A549 腺癌细胞系(A549(ASCL1+)细胞系)和 A549(对照)细胞系之间进行。RNA-seq 分析显示,H69 和 H69AR 细胞系之间有 940 个基因显著不同,A549(ASCL1+)和 A549(对照)细胞系之间有 728 个基因显著不同。总共,在这些分析中选择了 120 个常见基因作为候选 ASCL1 相关基因,这些基因具有各种细胞功能,如神经发育、分泌、生长和形态。对三种经典和两种变异 SCLC 细胞系、两种 A549(ASCL1+)和两种 A549(对照)细胞系中的这些候选基因的表达程度进行了定量 PCR 分析。由于候选 ASCL1 相关基因在经典 SCLC 和 A549(ASCL1+)细胞系中强烈表达,而在变异 SCLC 和 A549(对照)细胞系中表达较弱,因此进一步选择了 ASCL1 相关的 7 个分子 INSM1、ISL1、SYT4、KCTD16、SEZ6、MS4A8 和 COBL。这些分子提示 A549(ASCL1+)具有多种功能:INSM1 和 ISL1 是与神经内分泌分化相关的转录因子,而 SYT4、KCTD16 和 SEZ6 可能与神经分泌功能有关,MS4A8 和 COBL 与细胞生长和形态有关。对这些 7 个分子进行了肺癌组织和 A549(ASCL1+)的异种移植瘤的免疫组织化学染色,它们在 ASCL1 阳性肿瘤组织中优先呈阳性染色。

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