Exosomal miRNA profiling in Bcl-w overexpressed U251cells. Exosomal miRNA profiling in Bcl-w overexpressed U251cells

Exosomes are cell membrane-derived endoplasmic reticulum measuring 30 to 120 nm, which are secreted from various cells including cancer cells, and consist of lipids, proteins, mRNAs, and microRNAs (miRNAs). Tumor-derived exosomes are involved in tumor progression by delivering various factors to neighboring cells and promoting intercellular communication. MiRNA is a non-coding RNA consisting of 20-24 nucleotides that binds to the 3'UTR region of the target gene with a complementary nucleotide sequence during gene expression to degrade target mRNA or inhibit translation into target protein. In our study, exosomal miRNA profiling was utilized as a crucial technique to analyze the mechanisms of intercellular communication in tumor malignancy Overall design: We isolated exosomes from the conditioned medium of U251 cells overexpressing vector and Bcl-w. RNA was extracted from isolated exosomes and performed on a miRNA microarray.

外泌体（exosomes）是直径为30~120 nm、源自细胞膜的内质网来源囊泡，可由包括癌细胞在内的多种细胞分泌，其组成成分为脂质、蛋白质、信使RNA（mRNA）以及微小RNA（miRNAs）。
肿瘤源性外泌体可通过向邻近细胞传递多种因子、促进细胞间通讯，参与肿瘤进展过程。
微小RNA（miRNA）是一类长度为20~24个核苷酸的非编码RNA，在基因表达过程中可通过互补核苷酸序列结合至靶基因的3'非翻译区（3'UTR），从而降解靶信使RNA（mRNA）或抑制其翻译为靶蛋白。
本研究采用外泌体微小RNA表达谱分析作为关键技术，以解析肿瘤恶性进程中的细胞间通讯机制。
实验整体设计：我们从过表达空载体与Bcl-w的U251细胞的条件培养基中分离外泌体，从分离得到的外泌体中提取RNA，并开展微小RNA芯片检测。