Erigoster B Targeting DECR1 Induces Ferroptosis of Breast Cancer Cells via Promoting Phosphatidylcholine - Arachidonic Acid Metabolism

Breast caner, the leading cause of death in woman, is profoundly affected by coopreative changes of molecular and pathological characters in fatty acids metabolism. Transcriptome sequencing was performed to investigate the role of DECR1 in modulating molecular changes of breast cancer cells, we used MDAMB231 and BT474 cell lines with stable knockdown of DECR1. RNA was isolated with TRIzol reagent according to the manufacturer instructions. Briefly, cells were lysed with TRIzol directly in the culture dish. The lysate was transferred to a microcentrifuge tube, and chloroform was added. After vigorous shaking and centrifugation, the aqueous phase containing RNA was collected and mixed with isopropanol to precipitate the RNA. Following centrifugation, the RNA pellet was washed with ethanol, air-dried, and resuspended in RNase-free water. The concentration and purity of the RNA were measured using NanoDrop. The transcriptome sequencing library was constructed using the RNA Library Prep Kit for Illumina. Next, the library was sequenced on the Illumina NovaSeq platform. Raw reads in fastq format were processed. Then, the read count of each gene was calculated by the featureCounts package.

乳腺癌是女性人群的首要致死病因，其发生发展深受脂肪酸代谢过程中分子与病理特征协同变化的深刻影响。为探究DECR1在调控乳腺癌细胞分子特征中的作用，本研究开展转录组测序（Transcriptome sequencing）实验，选用两株稳定敲低DECR1的细胞系：MDA-MB-231与BT474。总RNA提取严格遵循TRIzol试剂说明书操作：将培养皿内的细胞直接以TRIzol裂解，随后将裂解液转移至微量离心管并加入氯仿；经充分振荡混匀与离心后，收集包含RNA的水相，加入异丙醇以沉淀RNA。离心完成后，用乙醇洗涤RNA沉淀，室温晾干后用无RNase水重悬RNA。采用NanoDrop分光光度计检测RNA的浓度与纯度；使用Illumina RNA文库制备试剂盒构建转录组测序文库，随后在Illumina NovaSeq测序平台完成上机测序。对fastq格式的原始测序reads进行预处理，再通过featureCounts软件包计算各基因的reads计数。