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.

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，我们在H69（ASCL1阳性，经典型SCLC）和H69AR（ASCL1阴性，变异型SCLC）之间以及在ASCL1之间进行了2个比较RNA-seq研究-转染的A549腺癌细胞系（A549（ASCL1 +）细胞系）和A549（对照）细胞系。RNA-seq分析显示，H69和H69AR细胞系之间有940个基因显着不同，而A549（ASCL1 +）和A549（对照）细胞系之间有728个基因显着不同。总共选择了这些分析之间的120个共同基因作为与ASCL1相关的候选基因，并包括具有多种细胞功能（如神经发育，分泌，生长和形态）的基因。对它们在三种经典和两种变异SCLC细胞系，两种A549（ASCL1 +）和两种A549（对照）细胞系中的表达程度进行定量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，而在变体SCLC和A549（对照）细胞系中较弱表达，还进一步选择了与ASCL1相关的7个分子，分别是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，KTCD16和SEZ6可能与神经分泌功能有关，而MS4A8和COBL可能与细胞生长和形态有关。这七个分子的免疫组织化学在肺癌组织和异种移植的A549（ASCL1 +）肿瘤上进行，并在ASCL1阳性肿瘤组织中被优先染色和阳性染色。SEZ6和SEZ6可能与神经分泌功能有关，MS4A8和COBL可能与细胞生长和形态有关。这七个分子的免疫组织化学在肺癌组织和异种移植的A549（ASCL1 +）肿瘤上进行，并在ASCL1阳性肿瘤组织中被优先染色和阳性染色。SEZ6和SEZ6可能与神经分泌功能有关，MS4A8和COBL可能与细胞生长和形态有关。这七个分子的免疫组织化学在肺癌组织和异种移植的A549（ASCL1 +）肿瘤上进行，并在ASCL1阳性肿瘤组织中被优先染色和阳性染色。