透過細胞分泌之細胞外囊泡內之信息傳遞者如微RNA，細胞間傳遞被認知為參與各種階段之腫瘤發展和轉移。因此本研究為探索癌細胞被非病毒性基因傅送後分泌之細胞外囊泡內之微RNA表現模式，以更進一步了解癌細胞經基因傳送後細胞間傳遞之分子信息資料。我們使用了二種常用非病毒性載體(lipofetamine 2000及jet polyethylenimine)來傳送綠螢光蛋白質體於HeLa 癌細胞，萃取純化分泌之細胞外囊泡並經次世代定序來分析其RNA組成。由卽時聚合酶鏈反應分析印證，我們發現二個共同重複表現於二種載體基因傳送之微RNAs(hsa-miR-143-3p和hsa-miR-193b-3p)，我們並預測其目標作用基因及相關之分子路徑分析，細胞死亡及壓力分子路徑之相關蛋白質表現亦一併分析印證。這二個微RNAs(hsa-miR-143-3p和hsa-miR-193b-3p)之上調節表現可被用於不同非病毒性載體基因傳送之癌細胞基因療法之潛在作用分子目標。 Intercellular communication is known to be involved in various stages of tumor development and metastasis through the secretion of extracellular vesicles (EVs) containing messengers such as microRNAs (miRNAs). Therefore, this study explored miRNA profiles in cancer cell-derived EVs after non-viral gene delivery in order to better understand the molecular information of intercellular communication in cancer cells after gene delivery. Two commonly used non-viral vectors (Lipofectamine 2000 and jet polyethylenimine) were used for the delivery of gene fluorescent protein plasmid in HeLa cancer cells. EVs were extracted and the contents of their RNA were subjected to the next-generation sequencing. In order to illustrate the common characteristics of non-viral vectors in the cancer cells, two overlapped up-regulated miRNAs (hsa-miR-143-3p and hsa-miR-193b-3p) were confirmed by real-time quantitative reverse transcriptase-polymerase chain reaction in the secreted EVs in response to both lipoplexes and polyplexes. The prediction of target genes and molecular pathways involved in these two miRNAs were determined and the protein expressions related to the pathways of cell death and stress in HeLa cells were identified. Hsa-miR-143-3p and hsa-miR-193b-3p were found to be up-regulated by the use of different non-viral vectors and can thus serve as potential targets of non-viral cancer gene therapy.
