MicroRNA Detection with Turnover Amplification via Hybridization-Mediated Staudinger Reduction for Pancreatic Cancer Diagnosis

Abstract

The occurrence of and development in the early pathological stage of pancreatic cancer has proved to be associated with microRNAs. However, it remains a great challenge to directly monitor low-expression, and downregulation of, microRNA among living cells, tissues, and serum samples. In this work, Staudinger reduction is first applied in intracellular microRNA detection, establishing a set of smart hybridization-mediated Staudinger reduction probes (HMSR-probe) which contain designed oligonucleotide sequences. Meanwhile, 40 serum samples (healthy people (6), patients with pancreatitis ( 22), and pancreatic cancer patients (12)) are tested for exploring the potential clinical application. Of note, the molecules bound to nucleic acid confine the reactive site to close proximity in a compact space, and nonconnected product from Staudinger reaction facilitates turnover amplification to an ameliorative detection limit (1.3 × 10 −15 M). Moreover, compared with qRT-PCR, a low false positive signal and an excellent specificity makes the probe more suitable and convenient for pancreatic cancer diagnosis in blood samples. For practical applications, HMSR-probe enable accurate differentiation in cell and tissue samples under both 488 and 785 nm and have good coherence to known research. As a proof of concept, the reliable results in distinguishing pancreatic cancer patients from different morbid stages might supply a feasible method for endogenous microRNA detection in fundamental research and clinical diagnostics.

Shedding Light on DNA‐Based Nanoprobes for Live‐Cell MicroRNA Imaging

“…Different from conventional stand-displacement mediated amplification reaction, Peng and co-worker first applied hybridization-mediated Staudinger reduction (HMSR) to design a fluorogenic response strategy for miRNA detection. [73] As shown in Figure 4E, the HMSR-probe was consisted with 1,8-naphthalimide derivative (DNA-NPAC) and 3-(diphenylphosphino) propionic acid (DNA-DPPA) to recognize and hybridize with target miRNA. It would confine the reactive site of azido group and diphenylphosphino molecule into proximity in a compact space, leading to the fluorescence recovery of the naphthalimide fluorophore.…”

Multicolor fluorescence encoding of different microRNAs in lung cancer tissues at the single-molecule level

“…3 Due to their simpler structure and less post-processing than DNA and protein, microRNAs (miRNAs) can function as the potential noninvasive biomarkers for early diagnosis and prognosis of lung cancer. [4][5][6][7][8][9] miRNAs are short endogenous non-coding RNAs with the length of ~22 nucleotide (nt). miRNAs can regulate the expression of specific genes by binding to complementary regions of messenger RNA (mRNA), and the resulting duplex may inhibit the translation by either enhancing the degradation or blocking the initiation of mRNA.…”

Dimensional Surface‐Enhanced Raman Scattering Nanostructures for MicroRNA Profiling

“…Therefore, the quantitation of single of multiple miRNA levels is of great significance for the assessment of human health, and can be applied to evaluate the different staging of cancer, the physical conditions of patients preoperatively, and prognosis. [10,11] Many profiling technologies [12][13][14][15][16][17][18][19][20][21] have already been applied to quantify miRNA levels; this work has made significant contributions in a range of molecular diagnostic tests in biomedicine. [22] Typically, quantitative real-time polymerase chain reaction (qRT-PCR) technology, [23] microRNA microarrays, [24] and northern blotting, [25] represent the gold standard tools for miRNAs profiling.…”