Establishment and Characterization of a Novel Pleuropulmonary Blastoma Cell Line

IF 3.1 2区医学 Q2 GENETICS & HEREDITY

Genes, Chromosomes & Cancer Pub Date : 2024-10-14 DOI:10.1002/gcc.23276

Keisuke Kato, Hiroaki Goto, Mio Tanaka, Tetsuomi Suzuki, Yasunori Toyoda, Masato Shinkai, Norihiko Kitagawa, Toshiji Nishi, Hisato Kigasawa, Kenji Kurosawa, Noriko Aida, Ai Yoshimi, Asami Noda, Yumi Ito, Masafumi Seki, Junko Takita, Noriyuki Nagahara, Masahiro Tsuchida, Yukichi Tanaka

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引用次数: 0

Abstract

Purpose

Pleuropulmonary blastoma (PPB) is an infrequently encountered childhood malignant intrathoracic neoplasm associated with unfavorable clinical behavior. Since a well-characterized preclinical model is essential for developing competent agents for PPB, we aim to establish and characterize the world's first cell line of PPB, and attempt to perform the cytotoxicity assay on the PPB cell line.

Experimental Design

The index case is a 2-year-old female who developed a right thoracic tumor that was surgically removed and treated with multi-agent chemotherapy. The patient is free from recurrence, although it was 9 years after the diagnosis when she developed a thyroid tumor. We performed in vitro cultivation of the isolated neoplastic cells from the tumor, cytogenetic findings and molecular analysis, and tetrazolium colorimetric assay.

Result

The histology was consistent with PPB. Serial passage of cultivation produced a continuously growing cell line, KCMC-PPB-1. Conventional cytogenetic analysis of the established cell line revealed complex numerical and structural chromosomal abnormalities, including add(17)(p11). Mutation analysis on the cultured cells revealed amino-acid substitution mutation on exon 4 of TP53 (NM_001276760.3:c.212_213delTG; NP_001263689.1:p.Leu72ArgfsTer37) and compound heterozygous mutations of DICER1 (NM_177438.3:c. 4910C>A; NP_803187.1:Ser1637* and NM_177438.3:c. 5114A>T; NP_803187.1:Glu1705Val). The cultivated cells demonstrated vulnerability to bortezomib on cytotoxicity assay.

Conclusion

Our KCMC-PPB-1 is the first genuine, molecularly characterized PPB cell line. The cell line is transplantable to nu/nu mice; therefore, it is suitable for a preclinical model for new drug development. The cytotoxicity assay demonstrated that bortezomib is active in the current PPB model.

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新型胸膜肺母细胞瘤细胞系的建立与特征描述

目的胸膜肺泡瘤（PPB）是一种罕见的儿童胸腔内恶性肿瘤，临床表现不佳。鉴于表征良好的临床前模型对于开发治疗 PPB 的有效药物至关重要，我们旨在建立世界上首个 PPB 细胞系并对其进行表征，同时尝试在 PPB 细胞系上进行细胞毒性试验。实验设计指标病例为一名 2 岁女性，患右侧胸部肿瘤，经手术切除并接受多种药物化疗。患者在确诊 9 年后又患上了甲状腺肿瘤，但至今没有复发。我们对从肿瘤中分离出的肿瘤细胞进行了体外培养、细胞遗传学检查和分子分析，以及四氮唑比色法检测。结果组织学与 PPB 一致。连续培养产生了一个持续生长的细胞系，即 KCMC-PPB-1。对已建立的细胞系进行常规细胞遗传学分析，发现了复杂的染色体数目和结构异常，其中包括 Add(17)(p11)。对培养细胞的突变分析表明，TP53 第 4 外显子上存在氨基酸替换突变（NM_001276760.3:c.212_213delTG; NP_001263689.1:p.Leu72ArgfsTer37），DICER1 存在复合杂合突变（NM_177438.3:c. 4910C>A; NP_001263689.1:p.Leu72ArgfsTer37）。4910C>A; NP_803187.1:Ser1637* 和 NM_177438.3:c.5114A>T; NP_803187.1:Glu1705Val).在细胞毒性试验中，培养的细胞显示出对硼替佐米的易感性。结论我们的 KCMC-PPB-1 是第一个真正的、具有分子特征的 PPB 细胞系。该细胞系可移植到 nu/nu 小鼠体内，因此适合作为新药开发的临床前模型。细胞毒性试验表明，硼替佐米在当前的 PPB 模型中具有活性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Genes, Chromosomes & Cancer 医学-遗传学

CiteScore

7.00

自引率

8.10%

发文量

审稿时长

4-8 weeks

期刊介绍： Genes, Chromosomes & Cancer will offer rapid publication of original full-length research articles, perspectives, reviews and letters to the editors on genetic analysis as related to the study of neoplasia. The main scope of the journal is to communicate new insights into the etiology and/or pathogenesis of neoplasia, as well as molecular and cellular findings of relevance for the management of cancer patients. While preference will be given to research utilizing analytical and functional approaches, descriptive studies and case reports will also be welcomed when they offer insights regarding basic biological mechanisms or the clinical management of neoplastic disorders.