FACS-based analysis of gene expression profile

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K09011
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Osaka University
• Principal Investigator: Takano Toru 大阪大学, 医学系研究科, 特任講師 (00263236)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: FACS / がん幹細胞 / 遺伝子発現プロフィール

Outline of Final Research Achievements

A single cell analysis using flow cytometry or fluorescence-activated cell sorting (FACS) is a useful tool to invesigate thyroid tissues. In this study, we selected early and mature thyroid markers, thyroid transcription factor 1 (TTF-1), thyroglobulin (TG), and sodium/iodine symporter (NIS), as well as N-cadherin as targets. Rat thyroid tissues and human thyroid cancer tissues were subjected for the following analysis. After 4-color analysis with FACS, gene expression profiles in FACS-purified cells were determined with microarrays. In rat thyroid tissues, three distinct cell populations showing different gene expresion profiles were identified. Three of 28 human thyroid cancer tissues showed clear heterogeniety in flow cytometry. In these samples, TG and NIS protein expression did not differ among cells, whereas TTF-1 and N-cadherin proteins exhibited heterogeneity. Using the present method, we identified distinct cell populations in both rat and human thyroid tissues.

Free Research Field

甲状腺疾患

Academic Significance and Societal Importance of the Research Achievements

幹細胞や癌幹細胞はその強力な増殖能から組織の再生に関与し、多くの疾患の経過に影響を与えている。しかし、これらは組織内に少数しか存在しないため同定や解析は極めて困難である。我々は、組織を単一細胞レベルまで分散し、RNAを保存した状態で固定し、蛍光標識抗体でラベルした後でFACSで目的の細胞を回収し、RNAを抽出・遺伝子発現プロフィールを解析することでそれらの性質を同定するFACS-mQという新しい方法を開発した。この方法を使用してラット・ヒト甲状腺組織を解析したところ、それぞれ特徴的な細胞分画の同定に成功した。当方法を用いることで疾患の新たな診断法が開発できる可能性がある。