利用低樣本高通量 MOWChIP 系統剖析紫杉醇對三陰性乳癌治療中組蛋白修飾的表觀遺傳機制

Abstract

乳癌為全球女性最常見的癌症類型，在台灣亦長年位居女性癌症死因之首，其中三陰性乳癌（Triple-negative breast cancer, TNBC）由於缺乏雌激素受體、黃體素受體與HER2表現的侵略性乳癌亞型，治療困難且預後較差。目前臨床主要依賴化學治療，其中以紫杉醇（Paclitaxel）為臨床上最廣泛使用之藥物，然而，其神經毒性的副作用嚴重且容易產生抗藥性。 本研究選用三陰性乳癌細胞株MDA-MB-231進行紫杉醇處理，並透過微流體染色質免疫共沉澱定序技術（MOWChIP-seq）分析給藥前後細胞中與活性增強子相關的H3K27ac組蛋白修飾。MOWChIP技術具備僅需微量樣本及高靈敏度捕捉優勢，能有效減少細胞使用量並提升免疫共沉澱效率。藉由比較紫杉醇處理前後的H3K27ac表觀遺傳圖譜，預期可鑑別出與藥物反應相關之基因調控變化。 本研究旨在探索Paclitaxel於三陰性乳癌細胞中引發的基因調控之變化，揭示其可能的表觀遺傳機制。可作為未來發展低毒性、具標靶性的表觀遺傳治療藥物之參考依據，提升治療三陰性乳癌的成效與安全性。

Breast cancer is the most common type of cancer among women worldwide and has long been the leading cause of cancer-related deaths among women in Taiwan. Among its subtypes, triple-negative breast cancer (TNBC), characterized by the absence of estrogen receptor (ER), progesterone receptor (PR), and HER2 expression, is highly aggressive, difficult to treat, and associated with poor prognosis. Chemotherapy remains the primary clinical treatment, with Paclitaxel being one of the most widely used drugs. However, its application is often limited by severe neurotoxic side effects and the development of drug resistance. In this study, the TNBC cell line MDA-MB-231 was treated with Paclitaxel, followed by epigenomic profiling using microfluidic oscillatory washing–based chromatin immunoprecipitation followed by sequencing (MOWChIP-seq) to examine changes in H3K27ac histone modifications associated with active enhancers before and after treatment. MOWChIP-seq enables highly sensitive ChIP-seq analysis with minimal cell input, improving enrichment efficiency while reducing sample consumption. By comparing genome-wide H3K27ac profiles before and after Paclitaxel treatment, we aim to identify transcriptional regulatory changes in response to the drug. This study seeks to explore the gene regulatory alterations induced by Paclitaxel in TNBC cells and to uncover the potential epigenetic mechanisms underlying its action. Our findings may provide a valuable reference for the development of novel epigenetic therapies with lower toxicity and greater specificity, thereby improving the safety and effectiveness of TNBC treatment.