Abstract
PurposeCurrently, the routine screening program has insufficient capacity for the early diagnosis of lung cancer. Therefore, a type of chitosan-molecular beacon (CS-MB) probe was developed to recognize the miR-155-5p and image the lung cancer cells for the early diagnosis.MethodsBased on the molecular beacon (MB) technology and nanotechnology, the CS-MB probe was synthesized self-assembly. There are four types of cells—three kinds of animal models and one type of histopathological sections of human lung cancer were utilized as models, including A549, SPC-A1, H446 lung cancer cells, tumor-initiating cells (TICs), subcutaneous and lung xenografts mice, and lox-stop-lox(LSL) K-ras G12D transgenic mice. The transgenic mice dynamically displayed the process from normal lung tissues to atypical hyperplasia, adenoma, carcinoma in situ, and adenocarcinoma. The different miR-155-5p expression levels in these cells and models were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The CS-MB probe was used to recognize the miR-155-5p and image the lung cancer cells by confocal microscopy in vitro and by living imaging system in vivo.ResultsThe CS-MB probe could be used to recognize the miR-155-5p and image the lung cancer cells significantly in these cells and models. The fluorescence intensity trends detected by the CS-MB probe were similar to the expression levels trends of miR-155 tested by qRT-PCR. Moreover, the fluorescence intensity showed an increasing trend with the tumor progression in the transgenic mice model, and the occurrence and development of lung cancer were dynamically monitored by the differen fluorescence intensity. In addition, the miR-155-5p in human lung cancer tissues could be detected by the miR-155-5p MB.ConclusionBoth in vivo and in vitro experiments demonstrated that the CS-MB probe could be utilized to recognize the miR-155-5p and image the lung cancer cells. It provided a novel experimental and theoretical basis for the early diagnosis of the disease. Also, the histopathological sections of human lung cancer research laid the foundation for subsequent preclinical studies. In addition, different MBs could be designed to detect other miRNAs for the early diagnosis of other tumors.
Highlights
Owing to late-stage detection and poor treatment, lung cancer is the leading cause of cancer-related deaths worldwide (Wang et al 2019a, b)
To determine whether the chitosan-molecular beacon (CS-molecular beacon (MB)) probe could detect the level of miRNA expression in viable cells, their ability to detect target microRNAs was assessed
Subcutaneous xenografts model, a: A549 treated with CS-RS MB, b:A549 treated with CS-miR-155-5p MB, c: H446 treated with CS-miR155-5p MB
Summary
Owing to late-stage detection and poor treatment, lung cancer is the leading cause of cancer-related deaths worldwide (Wang et al 2019a, b). The high mortality of lung cancer is always closely associated with late diagnosis (Zhang et al 2019a, b). Tumor-initiating cells (TICs) are considered as the source of tumor occurrence, development, recurrence, and metastasis (De et al 2018; Moro et al 2015; Yu et al 2016), thereby finding the TICs may be a novel breakthrough for the early diagnosis of cancers. If the TICs of lung cancer can be recognized and imaged directly based on the fluorescent substances, the early diagnosis can be implemented. Recognition and imaging of miRNAs related to lung cancer cells or TICs might constitute a critical strategy for the early diagnosis of the disease. MiR-155 plays a major role in the occurrence, development, and diagnosis of lung cancer (Zhang et al 2018; Shao et al 2019)
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