裂殖酵母中不稳定遗传耐药菌株的筛选及其运用
收稿日期: 2024-10-14
修回日期: 2024-12-18
网络出版日期: 2025-01-07
基金资助
国家自然科学基金项目(32400477);四川省自然科学基金青年基金(2023NSFSC1231)
Screening and application of unstable genetically resistant strains in fission yeast
Received date: 2024-10-14
Revised date: 2024-12-18
Online published: 2025-01-07
Supported by
National Natural Science Foundation of China(32400477);Natural Science Foundation of Sichuan Province(2023NSFSC1231)
发生在DNA序列水平或表观遗传水平的可逆遗传突变可以调控不稳定遗传的表型，其可逆性使得生物体能更好更快地适应外界多变的环境，但也正是因为其遗传的不稳定性，在传统研究(尤其是针对耐药性调控的研究)中往往被忽略。本研究利用雷帕霉素(+咖啡因)对野生型裂殖酵母菌株进行了耐药突变株的分离，共得到173株耐药突变株，经传代培养和子代耐药性试验，发现14株耐药株存在遗传不稳定现象。进一步研究表明，其中部分菌株的不稳定遗传耐药性受到ssp1位点可逆的DNA序列改变调控。本研究对雷帕霉素作为临床抗肿瘤药物治疗过程中易产生不稳定耐药性的分子机制提供了新的思路和相关的科学依据，并为解决其耐药性问题提供了可能的新作用靶点。
但露凤, 褚以文, 王欣荣, 何向伟 . 裂殖酵母中不稳定遗传耐药菌株的筛选及其运用[J]. 遗传, 2025 , 47(5) : 589 -599 . DOI: 10.16288/j.yczz.24-266
Reversible alterations at DNA sequence or epigenetic levels can result in phenotypes that are unstably inherited. The reversibility of these inheritable changes might be uniquely beneficial for adaption to possible fluctuations in environment. However, unstable changes are always ignored for the genetic instability in traditional studies, especially in the cause of drug resistance. In this study, we conduct a specific genetic screen in fission yeast using rapamycin (+caffeine) and obtain 173 resistant isolates. In contrast to the common strategy of isolating stable genetic mutants, we passage the cell culture with rapamycin resistance on drug free condition and test the resistance of offspring every five days, and obtain 14 strains that exhibit unstable resistance to rapamycin (the drug resistance is lost randomly among the cell progenies without drug selection pressure). Further studies show that the unstable genetic resistance of some strains is regulated by reversible DNA sequence alterationat the ssp1 gene locus. This study provides new insights and relevant scientific basis for the regulatory mechanism of unstable drug resistance in the process of rapamycin as a clinical anti-tumor drug, and a new possible target for solving the problem of drug resistance.
Key words： drug resistance; genetic instability; reversible mutation; rapamycin
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