in vivoエストロゲン非依存性乳癌細胞株による内分泌療法抵抗性機序の解析研究

使用体内雌激素非依赖性乳腺癌细胞系进行内分泌治疗耐药机制的分析研究

• 批准号: 22K08727
• 负责人: 山田 英幸
• 金额: $ 2.66万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2022
• 资助国家: 日本
• 起止时间: 2022-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22K08727/
• 关键词: Luminal type; エストロゲン抵抗性; breast cancer; animal model; endocrine therapy; estrogen signal

研究計画調書で示した通り、エストロゲン非依存性の乳癌細胞株(in vivo-estrogen-independent breast cancer celllines: iEIBCCs)を複数樹立した。それぞれの細胞株に対して免疫染色、WB (Western blotting)を行った。その結果ER (Estrogen Receptor)の発現や他の増殖シグナルの活性化については親株より発現が上昇しているものや同等のもの、低下しているものなど多様であり、各々異なるエストロゲン非依存性メカニズムを獲得していると考えられた。今回、我々は乳癌のエストロゲン依存性メカニズムの探索を目的としたiEIBCCsのRNA seqを施行し、それぞれの細胞株の増殖シグナル等を網羅的に解析した。RNA seqは親株、iEIBCCs　4種類、合わせて5種類の細胞株を用いtriplicateした。RNA seqの結果の主成分分析により評価を行ったところ、各celllinesの発現パターンの類似性から信頼度の高い解析結果であることが確認された。さらに、4種類のiEIBCCは全て親株と異なる性質を持ち、iEIBCCsもそれぞれ異なる遺伝子発現パターンを示し、これらは異なるER非依存性メカニズムを有していることを見出した。例えば、ERをコードする遺伝子のESR1の発現に着目したところ、親株と比較しESR1の発現が高度に失われた細胞株、中等度に失われた細胞株、発現が同等の細胞株、親株と比してESR1の発現を強く認めるものと多様であった。また、増殖シグナルの一つであるHER familyをコードする遺伝子 (ERBB2, ERBB3, ERBB4)についても同様であった。これらはWBにより、タンパク発現と相関することを確認している。

The research plan shows that the cell lines of non-dependent breast cancer (in vivo-estrogen-independent breast cancer celllines: iEIBCCs) are multiplicative. Cell lines were stained by immunostaining and WB (Western blotting) was detected by immunostaining. Results the results of ER (Estrogen Receptor) showed that there was no significant difference in the activity of other genes. The results showed that there was no significant difference between the two groups. Results: the results showed that there was no significant difference between the control group and the control group. Results: the results showed that there were significant differences between the two groups. The results showed that there was no significant difference between the control group and the control group. Results: the results showed that there was no significant difference between the two groups. This time, we are interested in breast cancer. We need to explore the implementation of iEIBCCs RNA seq and the analysis of cell lines, colonies, and other Internet sites. The cell strains of RNA seq strain, iEIBCCs 4 strain and iEIBCCs 5 strain were used for cell culture. RNA seq results the principal component analysis (PCA) was successful, and the celllines data showed that the data were similar and reliable. The results were highly analyzed. The full range of iEIBCC and 4 types of viruses has a positive response, the iEIBCCs has a negative response, and the ER is not dependent on each other. For example, ER, ESR1 and ESR1 showed that the height of the cell was higher than that of the ESR1, the height of the cell was higher than the height of the cell, the cell line of the moderate cell, the cell line of the same cell, the cell line of the same cell, the cell line of the cell line, the cell line of the cell line, the cell HER family, ERBB3, ERBB4, ERBB2, ERBB3, and so on. Please tell me that you have WB problems and that you have confirmed that you are not.