Quantitative Analysis of Multiple Proteins of Different Invasive Tumor Cell Lines at the Same Single‐Cell Level

Zhang, Xiangchun; Liu, Ru; Shu, Qingming; Yuan, Qing; Xing, Gengmei; Gao, Xueyun

doi:10.1002/smll.201703684

Cited by 24 publications

(24 citation statements)

References 56 publications

(34 reference statements)

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“…Single‐cell analysis tools report biomolecular heterogeneity that drives processes from immune‐cell response to cancer progression . Molecular standards play an important role in deconvolving biological variation from technical variation arising from a measurement tool .…”

Section: Introductionmentioning

confidence: 99%

Microparticle Delivery of Protein Markers for Single‐Cell Western Blotting from Microwells

Kim

Chan

Vlassakis

et al. 2018

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Immunoblotting confers protein identification specificity beyond that of immunoassays by prepending protein electrophoresis (sizing) to immunoprobing. To accurately size protein targets, sample analysis includes concurrent analysis of protein markers with known molecular masses. To incorporate protein markers in single-cell western blotting, microwells are used to isolate individual cells and protein marker-coated microparticles. A magnetic field directs protein-coated microparticles to >75% of microwells, so as to 1) deliver a quantum of protein marker to each cell-laden microwell and 2) synchronize protein marker solubilization with cell lysis. Nickel-coated microparticles are designed, fabricated, and characterized, each conjugated with a mixture of histidine-tagged proteins (42.3–100 kDa). Imidazole in the cell lysis buffer solubilizes protein markers during a 30 s cell lysis step, with an observed protein marker release half-life of 4.46 s. Across hundreds of individual microwells and different microdevices, robust log-linear regression fits (R2 > 0.97) of protein molecular mass and electrophoretic mobility are observed. The protein marker and microparticle system is applied to determine the molecular masses of five endogenous proteins in breast cancer cells (GAPDH, β-TUB, CK8, STAT3, ER-α), with <20% mass error. Microparticle-delivered protein standards underpin robust, reproducible electrophoretic cytometry that complements single-cell genomics and transcriptomics.

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Section: Introductionmentioning

confidence: 99%

Microparticle Delivery of Protein Markers for Single‐Cell Western Blotting from Microwells

Kim

Chan

Vlassakis

et al. 2018

Small

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“…The preparation method of Au clusters was similar to the method we previously reported [ 39 , 40 ]. The peptide (H 2 N-CCY-XX-HWKHLHNTKTFL-COOH) contains three domains.…”

Section: Resultsmentioning

confidence: 99%

Qualitative and Quantitative Analysis of Tumor Cell Invasion Using Au Clusters

et al. 2021

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Tumor invasion/metastasis is still the major cause of death in cancer patients. Membrane type-1 matrix metalloproteinase (MT1-MMP) is directly related to tumor invasion/metastasis. To accurately and quickly distinguish the risk of invasion/metastasis of primary tumor cells, it is urgent to develop a simple and precise quantitative method to distinguish the expression level of MT1-MMP. In this work, we have constructed red fluorescent Au clusters with peroxidase-like properties that could specifically bind to MT1-MMP on human cervical cancer cells. After MT1-MMP was labelled with Au clusters, we could visually see red fluorescence of MT1-MMP on cervical cancer cells via fluorescence microscopy and catalytic color imaging using an ordinary optical microscope. The constructed Au clusters contained 26 Au atoms; thus, the amount of MT1-MMP on cervical cancer cells could be accurately quantified using inductively coupled plasma mass spectrometry (ICP-MS). More importantly, the invasion/metastasis capabilities of the cervical cancer Siha, Caski and Hela cells with different MT1-MMP amounts could be accurately distinguished by fluorescence/catalysis qualitative imaging and ICP-MS quantitative analysis. This method of qualitative/quantitative analysis of tumor-associated proteins on cancer cells has great potential for accurately diagnosing aggressive tumor cells and assessment of their invasion/metastasis risk.

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“…Prior to exploring the cellular toxicity of the clusters and the cytotoxicological mechanism, we investigated the intracellular localization of AuCs on two tumor cells lines. Human primary glioblastoma cell line (U87-MG cells) is expressed with high level of intracellular H 2 O 2 and high expression of integrin α ν β 3 , while human cervical cancer cell line (HeLa cells) was expressed with low level of intracellular H 2 O 2 and low expression of integrin α ν β 3 [27,28]. The AuCs are co-cultured with above two mentioned cells lines for 12 h and then observed by confocal laser scanning microscopy (CLSM).…”

Section: Resultsmentioning

confidence: 99%

Peptide-Templated Gold Clusters as Enzyme-Like Catalyst Boost Intracellular Oxidative Pressure and Induce Tumor-Specific Cell Apoptosis

Zhang

Yuan

et al. 2018

Nanomaterials

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Anticancer metallodrugs that aim to physiological characters unique to tumor microenvironment are expected to combat drug tolerance and side-effects. Recently, owing to the fact that reactive oxygen species’ is closely related to the development of tumors, people are committed to developing metallodrugs with the capacity of improving the level of reactive oxygen species level toinduce oxidative stress in cancer cells. Herein, we demonstrated that peptide templated gold clusters with atomic precision preferably catalyze the transformation of hydrogen peroxide into superoxide anion in oxidative pressure-type tumor cells. Firstly, we successfully constructed gold clusters by rationally designing peptide sequences which targets integrin ανβ3 overexpressed on glioblastoma cells. The superoxide anion, radical derived from hydrogen peroxide and catalyzed by gold clusters, was confirmed in vitro under pseudo-physiological conditions. Then, kinetic parameters were evaluated to verify the catalytic properties of gold clusters. Furthermore, these peptide decorated clusters can serve as special enzyme-like catalyst to convert endogenous hydrogen peroxide into superoxide anion, elevated intracellular reactive oxygen species levels, lower mitochondrial membrane potential, damage biomacromolecules, and trigger tumor cell apoptosis consequently.

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Quantitative Analysis of Multiple Proteins of Different Invasive Tumor Cell Lines at the Same Single‐Cell Level

Cited by 24 publications

References 56 publications

Microparticle Delivery of Protein Markers for Single‐Cell Western Blotting from Microwells

Microparticle Delivery of Protein Markers for Single‐Cell Western Blotting from Microwells

Qualitative and Quantitative Analysis of Tumor Cell Invasion Using Au Clusters

Peptide-Templated Gold Clusters as Enzyme-Like Catalyst Boost Intracellular Oxidative Pressure and Induce Tumor-Specific Cell Apoptosis

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