PDF(Pubmed)
Abstract：
Ovarian cancer (OC) is one of the most lethal gynecologic cancers that is typically diagnosed at the very late stage of disease progression. Thus, there is an unmet need to develop diagnostic probes for early detection of OC. One approach may rely on RNA as a molecular biomarker. In this regard, FLJ22447 lncRNA is an RNA biomarker that is over-expressed in ovarian cancer (OC) and in cancer-associated fibroblasts (CAFs). CAFs appear early on in OC as they provide a metastatic niche for OC progression. FIT-PNAs (forced intercalation-peptide nucleic acids) are DNA analogs that are designed to fluoresce upon hybridization to their complementary RNA target sequence. In recent studies, we have shown that the introduction of cyclopentane PNAs into FIT-PNAs (cpFIT-PNA) results in superior RNA sensors. Herein, we report the design and synthesis of cpFIT-PNAs for the detection of this RNA biomarker in living OC cells (OVCAR8) and in CAFs. cpFIT-PNA was compared to FIT-PNA and the cell-penetrating peptide (CPP) of choice was either a simple one (four L-lysines) or a CPP with enhanced cellular uptake (CLIP6). The combination of CLIP6 with cpFIT-PNA resulted in a superior sensing of FLJ22447 lncRNA in OVCAR8 cells as well as in CAFs. Moreover, incubation of CLIP6-cpFIT-PNA in OVCAR8 cells leads to a significant decrease (ca. 60%) in FLJ22447 lncRNA levels and in cell viability, highlighting the potential theranostic use of such molecules.
摘要：
卵巢癌（OC）是最致命的妇科癌症之一，通常在疾病进展的晚期被诊断出来。因此，开发用于早期检测OC的诊断探针的需求尚未满足。一种方法可以依赖于RNA作为分子生物标志物。在这方面,FLJ22447lncRNA是在卵巢癌(OC)和癌症相关成纤维细胞(CAF)中过表达的RNA生物标志物。CAF在OC中早期出现，因为它们为OC进展提供了转移性生态位。FIT-PNA(强制嵌入肽核酸)是被设计为在与其互补RNA靶序列杂交时发出荧光的DNA类似物。在最近的研究中,我们已经表明，将环戊烷PNA引入FIT-PNA（cpFIT-PNA）可产生出色的RNA传感器。在这里，我们报道了cpFIT-PNA的设计和合成，用于检测活OC细胞（OVCAR8）和CAF中的这种RNA生物标志物。将cpFIT-PNA与FIT-PNA进行比较，选择的细胞穿透肽（CPP）是简单的肽（四个L-赖氨酸）或具有增强细胞摄取的CPP（CLIP6）。CLIP6与cpFIT-PNA的组合导致在OVCAR8细胞以及CAFs中的FLJ22447lncRNA的优异感测。此外，在OVCAR8细胞中孵育CLIP6-cpFIT-PNA导致显着降低（约60%)在FLJ22447lncRNA水平和细胞活力中，强调了这种分子的潜在治疗用途。
