Detection of mRNAs of Ribosomal Protein L15 and E-Cadherin in Living Circulating Tumor Cells at Single Cell Resolution To Study Tumor Heterogeneity

Xu, Chi; He, Xiaoyan; Ren, Xiaohe; Han, Di; Cheng, Si‐Xue

doi:10.1021/acs.analchem.2c00488

Cited by 5 publications

(5 citation statements)

References 50 publications

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“…RPs are widely distributed in various tissues, and have specific transcript expression profiles in normal human tissues (Bortoluzzi et al, 2001). Recent studies showed that the expression patterns of RPs were related to various pathological conditions (Sun et al, 2022; Xu, He, et al, 2022). Multiple RPs have been found to be differentially expressed in many types of tumors, such as colorectal cancer, prostate cancer, and breast cancer (Azevedo et al, 2022; Bee et al, 2006; Markiewski et al, 2017).…”

Section: Discussionmentioning

confidence: 99%

Decreased expression of RPL15 and RPL18 exacerbated the calcification of valve interstitial cells during aortic valve calcification

Wang

Qin

Zhang

et al. 2023

Cell Biology International

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Calcific aortic valve disease (CAVD) is the most common valvular heart disease, with an increasing prevalence due to an aging population. The pathobiology of CAVD is a multifaceted and actively regulated process, but the detailed mechanisms have not been elucidated. The present study aims to identify the differentially expressed genes (DEGs) in calcified aortic valve tissues, and to analyze the correlation between DEGs and clinical features in CAVD patients. The DEGs were screened by microarray in normal and CAVD groups (n = 2 for each group), and confirmed by quantitative real‐time polymerase chain reaction in normal (n = 12) and calcified aortic valve tissues (n = 34). A total of 1048 DEGs were identified in calcified aortic valve tissues, including 227 upregulated mRNAs and 821 downregulated mRNAs. Based on multiple bioinformatic analyses, three 60S ribosomal subunit components (RPL15, RPL18, and RPL18A), and two 40S ribosomal subunit components (RPS15 and RPS21) were identified as the top 5 hub genes in the protein–protein interaction network of DEGs. The expression of RPL15 and RPL18 was also found significantly decreased in calcified aortic valve tissues (both p < .01), and negatively correlated with the osteogenic differentiation marker OPN in CAVD patients (both p < .01). Moreover, inhibition of RPL15 or RPL18 exacerbated the calcification of valve interstitial cells under osteogenic induction conditions. The present study proved that decreased expression of RPL15 and RPL18 was closely associated with aortic valve calcification, which provided valuable clues to find therapeutic targets for CAVD.

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Section: Discussionmentioning

confidence: 99%

Decreased expression of RPL15 and RPL18 exacerbated the calcification of valve interstitial cells during aortic valve calcification

Wang

Qin

Zhang

et al. 2023

Cell Biology International

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“…As a type of oligonucleotide-based sensor recognizing specific nucleic acid sequences, MBs can realize sensitive detection of nucleic acids such as mRNAs and miRNAs. Being delivered into living cells by a delivery vector, MBs can probe intracellular RNA targets in situ at a single cell resolution. − …”

Section: Introductionmentioning

confidence: 99%

Precise Detection on Cell–Cell Fusion by a Facile Molecular Beacon-Based Method

Ren

Han

et al. 2022

Anal. Chem.

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Cell−cell fusion studies provide an experimental platform for evaluating disease progression and investigating cell infection. However, to realize sensitive and quantitative detection on cell−cell fusion is still a challenge. Herein, we report a facile molecular beacon (MB)-based method for precise detection on cell−cell fusion. By transfection of the spike protein (S protein) and enhanced green fluorescent protein (EGFP) in HEK 293 cells, the virus-mimicking fusogenic effector cells 293-S-EGFP cells were constructed to interact with target cells. Before mixing the effector cells with the target cells, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in 293-S-EGFP cells was silenced, and the MB for GAPDH mRNA detection was delivered into the GAPDH silenced 293-S-EGFP cells. Once cell−cell fusion occurred, MB migrated from the GAPDH silenced effector cells to the target cells and hybridized with GAPDH mRNA in the target cells to induce fluorescence emission. The cell−cell fusion can be easily visualized and quantitated by fluorescence microscopy and flow cytometry. The fluorescence intensity is strongly dependent on the number of fused target cells. This MB-based method can easily identify the differences in the cell fusions for various target cells with different angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) expression levels, resulting in dramatically different fluorescence intensities in fused target cells. Our study provides a convenient and efficient quantitative detection approach to study cell−cell fusion.

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“…[20,21] Aptamers conjugated on HA are introduced to the delivery system via electrostatic interactions. The SYL3C aptamer can specifically interact with epithelial cell adhesion molecule (EpCAM) overexpressed on epithelial cancer cells, [10,22] and the CL4 aptamer can bind epidermal growth factor receptor (EGFR) overexpressed on mesenchymal cancer cells. [23] The delivery system can target all CTCs during EMT and MET processes.…”

Section: Introductionmentioning

confidence: 99%

“…To address this issue, in our previous work, we developed tumor-targeting nano-systems for exerting therapeutic actions and/or probing biomarkers in CTCs. [9][10][11] As it is well known, epithelial-mesenchymal transition (EMT) and stemness are critical players in cancer progression. [12] Diverse subtyped CTCs, including CTCs in different EMT and mesenchymal-epithelial transition (MET) stages, and CTCs with varied stemness levels, may co-exist.…”

Section: Introductionmentioning

confidence: 99%

Multi‐Targeting Nano‐Systems Targeting Heterogeneous Cancer Cells for Therapeutics and Biomarker Detection

Ren

Han

et al. 2022

Adv Healthcare Materials

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Cancer heterogeneity plays a vital part in cancer resistance and metastasis. To provide a reliable approach to exert a therapy action and evaluate its efficiency in heterogeneous cancer cells, a multiple targeting delivery vector composed of histone encapsulating the therapeutic or diagnostic agent, hyaluronic acid targeting CD44 overexpressed in stem tumor cells, SYL3C aptamer targeting epithelial cell adhesion molecule (EpCAM) overexpressed in epithelial cancer cells, and CL4 aptamer targeting epidermal growth factor receptor (EGFR) overexpressed in mesenchymal cancer cells, is developed. The vector can efficiently target different cancer cells and circulating tumor cells (CTCs) in the peripheral blood of patients for mucin 1 (MUC1) knockout. Furthermore, the multiple targeting vector can be used to co-encapsulate three types of molecular beacons for probing various mRNA biomarkers at single-cell resolution after genome editing. This study provides an efficient approach for exerting therapeutic actions in heterogeneous cancer cells and assessing the therapeutic efficacy by detection of cancer biomarkers via liquid biopsy.

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Detection of mRNAs of Ribosomal Protein L15 and E-Cadherin in Living Circulating Tumor Cells at Single Cell Resolution To Study Tumor Heterogeneity

Cited by 5 publications

References 50 publications

Decreased expression of RPL15 and RPL18 exacerbated the calcification of valve interstitial cells during aortic valve calcification

Decreased expression of RPL15 and RPL18 exacerbated the calcification of valve interstitial cells during aortic valve calcification

Precise Detection on Cell–Cell Fusion by a Facile Molecular Beacon-Based Method

Multi‐Targeting Nano‐Systems Targeting Heterogeneous Cancer Cells for Therapeutics and Biomarker Detection

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