Different Types Of Tumor Cells Research Articles

Microdroplet-SERS platform for single cell-secreted VEGF and extracellular pH analysis in oxidative stress event

Although oxidative stress processes based on reactive oxygen species have been the research focus, analyzing this phenomenon at the single-cell level is challenging. A microfluidic platform combined with highly sensitive surface-enhanced Raman scattering (SERS) technology was constructed. The secretion of the vascular endothelial growth factor (VEGF) and the extracellular microenvironmental pH fluctuations of a single cell during the oxidative stress process were assessed and analyzed. The immune sandwich structure formed between the capture probe and the reporter probe with the linkage of secreted VEGF was built above the probed cell surface in each drop. The self-driven collection behavior of the magnetic bead-based capture probe toward the cell surface significantly amplified the SERS signals of the reporter probe, thus improving the sensitivity and accuracy of detection. 4-Mercaptopyridine (4-Mpy) responding sensitively to pH was applied to label the reporter probe, which endows the extracellular microenvironmental pH sensing during oxidative stress events according to relative SERS intensity. Combined with machine learning to analyze the spectral characteristics, the distinction between normal cells and different types of tumor cells was finally realized. This study makes full use of the advantages of machine learning to process spectral data to explore deep information and provides an auxiliary tool for diagnosing cancer and other medical conditions in the future.

Isolation, culture and identification of brain tumor stem cells from glioma

Objectives: This study aims to provide a feasible method for culturing tumor stem cells to facilitate further research on the occurrence and development of brain tumors, making it applicable for in vitro studies of brain tumor stem cells. Methods: Samples of glioma were collected and subsequently isolated and cultured, then immunocytochemical staining, including CD133, Nestin, and GFAP staining, was performed on the cultured cells. Results: Some of the primary cultured tumor stem cells grew in the shape of sphere and formed into clusters. With the extension of culture time, the cell mass gradually increased, with strong proliferation and self-renewal ability. Meanwhile, the tumor derived spherical cells could differentiate into different types of tumor cells in serum-containing medium since most of them were positive to CD133, Nestin, and GFAP on the 7 days. Conclusion: The tumor stem cell can differentiate into different cell types, carrying significant implications for tumor research and treatment. More importantly, the methods introduced in this study are simple and feasible to isolate and culture tumor stem cell, contributing to cell source for future studies.

Combining mRNA with PBS and calcium ions improves the efficiency of the transfection of mRNA into tumors

Messenger RNA (mRNA) is a promising modality for expressing a protein in vivo. Drug delivery systems are required for the efficient transfection of mRNA into cells. In this study, we evaluated several drug delivery systems for transfecting mRNA into tumors. A lipid nanoparticle delivered mRNA to the draining lymph nodes and liver, even by intratumoral injection. A liposome-based system did not consistently provide mRNA for different types of tumor cells. We found that phosphate-buffered saline (PBS) introduced mRNA into several tumors, and calcium ions enhanced the efficiency, particularly in male mice. The circular dichroism spectrometer suggested a structural change in mRNA in PBS. Transmission electron microscopy revealed that calcium ions promoted the formation of mRNA nanoparticles in PBS. Transfection of mRNAs coding OX40-ligand, interleukin 36γ, and interleukin 23 by PBS + calcium ions attenuated tumor growth. Our results indicate that combining PBS with calcium ions promotes the transfection of mRNA into tumors. These data provide information for the development of methods for transfection of mRNA for cancer therapy.

Nanozyme-Inhibited SERS Multichannel Paper-Based Sensor Array for the Quantification and Identification of Biothiols and Cancer Cells Based on Three Ag-Based Nanomaterials.

Biothiols play essential roles in maintaining normal physiological functions, resisting oxidative stress, and protecting cell health. Establishing an effective and reliable sensor array for the accurate quantification and discrimination of diverse biothiols is extremely meaningful. In this work, Ag/Mn3O4, Ag3PO4, and Ag3Cit with excellent oxidase-mimetic activity and surface-enhanced Raman scattering (SERS)-enhanced features have been prepared and loaded onto Whatman filter paper (WFP) to build SERS paper chips as three sensing channels, which can induce 3,3',5,5'-tetramethylbenzidine (TMB) oxidation to SERS-active reporters (TMBox) and concurrently generate prominent SERS signals. Nevertheless, the addition of biothiols can suppress conversion from TMB to TMBox, which can cause the reduction of the SERS signal from TMBox. Interestingly, each SERS sensing channel can generate different TMBox signals' variations due to differences in the oxidative inhibition abilities of diverse biothiols and exclusive properties of each paper chip, which can be plotted as specific fingerprint patterns of each biothiol and further translated into intuitive two-dimensional (2D) clustering profiles through linear discriminant analysis (LDA) and hierarchical cluster analysis (HCA) techniques for precise identification of these six biothiols with the minimum concentration of 1 μM. More importantly, this SERS sensor array is exploited for the precise quantification of intracellular glutathione (GSH), and can differentiate between normal and cancer cells based on different intracellular GSH contents and even identify different types of tumor cells, demonstrating its powerful application prospects in disease diagnosis.

Quinalizarin as a potential antifungal drug for the treatment of Candida albicans fungal infection in cancer patients.

This study aims to analyze the antifungal properties of quinalizarin, a plant-derived compound with proven anticancer effects. Quinalizarin exhibited antifungal activity against opportunistic pathogenic Candida species and Geotrichum capitatum. The treatment with this anthraquinone reduced hyphal growth, inhibited biofilm formation, and damaged mature Candida albicans biofilms. Real-time RT-PCR revealed that quinalizarin downregulated the expression of hyphae-related and biofilm-specific genes. The flow cytometry method used in the study showed that both apoptosis and necrosis were the physiological mechanisms of quinalizarin-induced C. albicans cell death, depending on the dose of the antifungal agent. A further study revealed an increase in the levels of intracellular reactive oxygen species and alterations in mitochondrial membrane potential after treatment with quinalizarin. Finally, quinalizarin was found to have low toxicity in a hemolytic test using human erythrocytes. In conclusion, we have identified quinalizarin as a potential antifungal compound.IMPORTANCEThis article is a study to determine the antifungal activity of quinalizarin (1,2,5,8-tetrahydroxyanthraquinone). Quinalizarin has potential antitumor properties and is effective in different types of tumor cells. The aim of the present study was to prove that quinalizarin can be used simultaneously in the treatment of cancer and in the treatment of intercurrent fungal infections. Quinalizarin was identified as a novel antifungal compound with low toxicity. These results may contribute to the development of a new drug with dual activity in the treatment of cancer-associated candidiasis.

Elevated expression patterns of P-element Induced Wimpy Testis (PIWI) transcripts are potential candidate markers for Hepatocellular Carcinoma.

P-Element-induced wimpy testis (PIWI) proteins, when in combination with PIWI-interacting RNA (piRNA), are engaged in the epigenetic regulation of gene expression in germline cells. Different types of tumour cells have been found to exhibit abnormal expression of piRNA, PIWIL-mRNAs, and proteins. We aimed to determine the mRNA expression profiles of PIWIL1, PIWIL2, PIWIL3, & PIWIL4, in hepatocellular carcinoma patients, and to associate their expression patterns with clinicopathological features. The expression patterns of PIWIL1, PIWIL2, PIWIL3, PIWIL4 mRNA, was assessed via real-time quantitative polymerase chain reaction (RT-QPCR), on tissue and serum samples from HCC patients, their impact for diagnosis was evaluated by ROC curves, prognostic utility was determined, and In Silico analysis was conducted for predicted variant detection, association with HCC microRNAs and Network Analysis. Expression levels were significantly higher in both HCC tissue and serum samples than in their respective controls (p< 0.001). Additionally, the diagnostic performance was assessed, Risk determination was found to be statistically significant. PIWIL mRNAs are overexpressed in HCC tissue and serum samples, the expression patterns could be valuable molecular markers for HCC, due to their association with age, tumour grade and pattern. To the best of our knowledge, our study is the first to report the expression levels of all PIWIL mRNA and to suggest their remarkable values as diagnostic and prognostic biomarkers, in addition to their correlation to HCC development. Additionally, a therapeutic opportunity might be also suggested through in silico miRNA prediction for HCC and PIWIL genes through DDX4 and miR-124-3p.

Homomultivalent Polymeric Nanotraps Disturb Lipid Metabolism Homeostasis and Tune Pyroptosis in Cancer.

Genetic manipulations and pharmaceutical interventions to disturb lipid metabolism homeostasis have emerged as an attractive approach for the management of cancer. However, the research on the utilization of bioactive materials to modulate lipid metabolism homeostasis remains constrained. In this study, heptakis (2,3,6-tri-O-methyl)-β-cyclodextrin (TMCD) is utilized to fabricate homomultivalent polymeric nanotraps, and surprisingly, its unprecedented ability to perturb lipid metabolism homeostasis and induce pyroptosis in tumor cells is found. Through modulation of the density of TMCD arrayed on the polymers, one top-performing nanotrap, PTMCD4, exhibits the most powerful cholesterol-trapping and depletion capacity, thus achieving prominent cytotoxicity toward different types of tumor cells and encouraging antitumor effects in vivo. The interactions between PTMCD4 and biomembranes of tumor cells effectively enable the reduction of cellular phosphatidylcholine and cholesterol levels, thus provoking damage to the biomembrane integrity and perturbation of lipid metabolism homeostasis. Additionally, the interplays between PTMCD4 and lysosomes also induce lysosomal stress, activate the nucleotide-binding oligomerization domain-like receptor protein 3 inflammasomes, and subsequently trigger tumor cell pyroptosis. To sum up, this study first introduces dendronized bioactive polymers to manipulate lipid metabolism and has shed light on another innovative insight for cancer therapy.

Learning parameter values of a fractional model of cancer employing boundary densities of tumor cells

In this paper, a free boundary model of tumor growth with drug application including the heterogeneity or different types of tumor cells (caused by mutations and different values of drug and nutrient concentrations inside the tumor) is studied. Heterogeneity is included in the model by a variable . It is assumed that converting from mutation to mutation happens with probability . A Caputo time fractional‐order hyperbolic equation describes the evolution of tumor cells depending on . It also includes two Caputo time fractional‐order parabolic equations describing the diffusions of nutrients (e.g., oxygen and glucose) and drug concentrations. Instead of integer‐order time derivatives, the fractional ones are considered. In this study, it is aimed to employ the least squares curve fitting method to fit the order of fractional derivatives, coefficient, and rates of the model. For this purpose, using the mathematical model, we have considered the boundary densities of the tumor cells of different types and near‐boundary concentrations of drug and nutrient as the functions of the unknown orders, coefficients and rates (unknown variables). Due to the complexity of the problem, we have obtained the functions numerically. For this, using a change of variable, we have changed the free boundary problem to a problem with fixed domain. Thus, Riemann–Liouville fractional‐order integrals are added to the problem. In the spatial domain, the problem is discretized using the collocation method. In the temporal domain, the fractional derivatives and integrals (with order ) are approximated in mesh points (with step size ) applying a method with error . Then, the unknown variables are obtained by fitting the functions for unknown variables to the data. In order to obtain the variables, a quadratic objective function is considered. The discretized function is substituted into the objective function, then the objective function is minimized using the trust‐region reflective algorithm. Finally, some numerical examples are presented to verify the efficiency of the method. In the examples, we have added noises generated from Gaussian distributions to the data and the effects of noise on the fitted coefficients, and rates are illustrated using some figures and tables. The noisy data are also plotted to have a clear vision of the effects of noises on the data. It is shown that despite the noise, the prediction of the radius of the tumor is acceptable.

Therapeutic Potential of Heat Shock Protein 90 Inhibitors, Geldanamycin, and Analog Compounds in Precision Cancer Therapy

Heat shock protein (HSP90) is a molecular chaperone involved in numerous physiological processes. The primary role of this is to assist in the process of protein folding and to restore misfolded proteins to their correct shape. Chaperones additionally inhibit protein breakdown and aggregation. HSP90 inhibitors possess a notable characteristic of obstructing many cancer-causing pathways by facilitating the breakdown of numerous oncogenic client proteins. Targeting HSP90 therapeutics has been recognized as a viable approach for treating cancer and inflammatory-associated disorders in clinical studies involving different forms of cancer. Inhibition of HSP90 using natural, synthetic, and semi-synthetic chemicals has shown encouraging outcomes. HSP90 inhibitors have been extracted from several fungi, bacteria, and plant species. These naturally occurring chemicals play a crucial function in regulating HSP90 activity and can be utilized to develop innovative semi-synthetic or synthetic inhibitors. Over 120 clinical trials have been carried out to evaluate the effectiveness of HSP90 inhibitors as a supplementary therapy for different types of tumor cells. Presently, ongoing research is being carried out to acquire an understanding of innovative and more efficacious methods for treating cancer. Continuing in this research approach, we aim to investigate the discovery, biosynthesis, mechanism of action, and biological features of geldanamycin and its analogs.

ALDH3A2 Regulates Ferroptosis By Participating in Fatty Acid Synthesis and Finally Lead to Chemotherapy Resistance in Acute Myeloid Leukemia

Chemotherapeutic resistance is a major challenge in the treatment of acute myeloid leukemia (AML). Although increasing the dose of anthracycline drugs improve chemotherapy efficacy and reduce drug resistance, the side effects limit the increase of their dosage. One strategy to improve the therapeutic effects is to improve the efficacy in their limited doses. Ferroptosis is one important way in which Adriamycin exerts cytotoxic effects on AML cells, which was also associated with resistance to Adriamycin. ALDH3A2 is a fatty aldehyde dehydrogenase related to the conversion of fatty aldehydes and fatty acids. Here, we found that ALDH3A2 is a biomarker for chemotherapy-resistant AML, increased expression of ALDH3A2 reduced ferroptosis and cytotoxicity induced by Adriamycin in AML cells. In our studies, we demonstrated that ALDH3A2 regulated ferroptosis and AML drug resistance by participating in fatty acid synthesis. By analyzing the data from TCGA and GTEx databases, we found that ALDH3A2 expression varied in different types of tumor cells. AML patients had higher ALDH3A2 expression than normal tissues. We analyzed the RNA-seq results of 88 leukemia cell lines from the HPA database, and found that 15 out of the top 25% highest ALDH3A2 expression cell lines were AML cell lines, indicating that AML cells had higher ALDH3A2 expression than other types of leukemia cells. We plotted survival curves based on the gene data and clinical survival data from TCGA and GTEx databases. It showed that AML patients with high ALDH3A2 expression had shorter overall survival (OS) than those with low expression. We detected the expression of ALDH3A2 in AML cell lines and primary cells from patients using Western blot and RT-qPCR, respectively. Our data showed that drug-resistant AML cell lines had higher ALDH3A2 expression than their parental cells. The expression of ALDH3A2 in primary cells from AML patients achieving continuous complete remission (CR) within two years was significantly lower than that in the first-line non-remission(C1NR) group and the relapsed/refractory (R/R) group. Besides, the expression of ALDH3A2 is related to certain types of mutation such as CEBPα, FLT3-ITD and RUNX1. To investigate the function of ALDH3A2, we constructed AML cell lines with ALDH3A2 knockdown or overexpression. We found that the increasing ferroptosis level and decreasing cell activity after treatment with the same concentration of Adriamycin was observed in the knockdown group, and overexpression group had the opposite effect. By ELISA and GC-MS, we discovered that there were differences in 4-HNE and fatty acid content between the overexpression group and the control group. It suggested that ALDH3A2 overexpression reduced the cytotoxic 4-HNE and increased the content of heptadecanoic acid, oleic acid, and linoleic acid. We further pretreated AML cells with heptadecanoic acid, oleic acid, and linoleic acid, and found that pretreatment with heptadecanoic acid and oleic acid reduced ferroptosis level and cytotoxicity induced by Adriamycin in AML cells. We also tried to find the regulatory mechanism of ALDH3A2 expression and found that acetylation plays a certain role in it. In conclusion, our studies found that increased expression of ALDH3A2 was related to poor chemotherapy efficacy in AML patients and clarifies the mechanism of ALDH3A2 regulating chemosensitivity in AML. ALDH3A2 degraded the cell toxic product 4-HNE of lipid peroxidation and regulated the proportion and content of heptadecanoic acid, oleic acid, and linoleic acid in cells, which related to ferroptosis and resistance to anthracycline drugs. Downregulating ALDH3A2 could increase AML's susceptibility to chemotherapy which may make it a promising therapy.

A Novel Optical Fiber Terahertz Biosensor Based on Anti-Resonance for The Rapid and Nondestructive Detection of Tumor Cells.

The sensitive and accurate detection of tumor cells is essential for successful cancer therapy and improving cancer survival rates. However, current tumor cell detection technologies have some limitations for clinical applications due to their complexity, low specificity, and high cost. Herein, we describe the design of a terahertz anti-resonance hollow core fiber (THz AR-HCF) biosensor that can be used for tumor cell detection. Through simulation and experimental comparisons, the low-loss property of the THz AR-HCF was verified, and the most suitable fiber out of multiple THz AR-HCFs was selected for biosensing applications. By measuring different cell numbers and different types of tumor cells, a good linear relationship between THz transmittance and the numbers of cells between 10 and 106 was found. Meanwhile, different types of tumor cells can be distinguished by comparing THz transmission spectra, indicating that the biosensor has high sensitivity and specificity for tumor cell detection. The biosensor only required a small amount of sample (as low as 100 μL), and it enables label-free and nondestructive quantitative detection. Our flow cytometry results showed that the cell viability was as high as 98.5 ± 0.26% after the whole assay process, and there was no statistically significant difference compared with the negative control. This study demonstrates that the proposed THz AR-HCF biosensor has great potential for the highly sensitive, label-free, and nondestructive detection of circulating tumor cells in clinical samples.

Development of potent isoflavone-based formyl peptide receptor 1 (FPR1) antagonists and their effects in gastric cancer cell models

Formyl peptide receptor-1 (FPR1) is a G protein-coupled chemoattractant receptor that plays a crucial role in the trafficking of leukocytes into the sites of bacterial infection and inflammation. Recently, FPR1 was shown to be expressed in different types of tumor cells and could play a significant role in tumor growth and invasiveness. Starting from the previously reported FPR1 antagonist 4, we have designed a new series of 4H-chromen-2-one derivatives that exhibited a substantial increase in FPR1 antagonist potency. Docking studies identified the key interactions for antagonist activity. The most potent compounds in this series (24a and 25b) were selected to study the effects of the pharmacological blockade of FPR1 in NCl–N87 and AGS gastric cancer cells. Both compounds potently inhibited cell growth through a combined effect on cell proliferation and apoptosis and reduced cell migration, while inducing an increase in angiogenesis, thus suggesting that FPR1 could play a dual role as oncogene and onco-suppressor.

TREX1 cytosolic DNA degradation correlates with autoimmune disease and cancer immunity.

Three Prime Repair Exonuclease 1 (TREX1) is a major 3'-5' exonuclease in mammalian cells that primarily degrades DNA substrates in the cytoplasm. TREX1 deficiency results in an aberrant accumulation of self-DNA in the cytosol, predominantly activating the innate immune response via the cGAS-STING-IFN pathways. Characterized by interferon production, autoimmune diseases, such as Aicardi-Goutieres syndrome (AGS), systemic lupus erythematosus (SLE), familial chilblain lupus (FCL), and retinal vasculopathy and leukodystrophy (RVCL), are associated with TREX1 loss of function. Besides, abnormal expression of TREX1 also appears in different types of tumor cells. In this review, we described the structure and mechanisms of TREX1 in degrading self-DNA and discussed the potential sources of these aberrant nucleic acids. We also summarized the major research advances in TREX1-related diseases, including autoimmune diseases and cancer. These contents will serve as a foundation for further study on the function of TREX1 and provide advances in the diagnosis, treatment, and prognosis of related disorders.

Utilizing endosomal capture for tumor therapy via membrane-lytic mechanism-based Pickering emulsion.

Nanocarriers are easily captured by endosomes, where the abundant hydrolases inevitably destroy the nanocarriers and the drugs they carry, ultimately resulting in a compromised or lost therapeutic efficacy. Herein, we report a membrane-lytic mechanism-based Pickering emulsion that can in turn utilize this seemingly unfavorable endosomal capture behavior for tumor therapy. This Pickering emulsion is constructed as an oil-in-water (O/W) emulsion stabilized by the hybrid nanoparticles (HNPs) composed of two molecules with opposite charges, cetyl trimethylamine bromide (CTAB) and linoleic acid (LA), through electrostatic interaction (defined as HNPs@PE). After HNPs@PE enters the lysosomes through macropinocytosis-mediated endocytosis, LA can be protonated in response to the acidic stimulus, and causing the swelling or disintegration of HNPs due to the disrupted electrostatic interaction. The released CTAB holds strong membrane-lytic activity and can directly damage the lysosomal membranes. Under the acidic condition and the participation of excessive iron ions (II) in lysosomes, LA induces lipid peroxidation and the resulting lipid peroxides (LPO) will oxidize the lysosomal membranes, collectively causing the leakage of lysosome membranes and the release of contents into cytoplasm. Subsequently, the diffused CTAB and LPO will continue to attack the mitochondrial membranes and cell membranes, resulting in the death of different types of tumor cells both in vitro and in vivo due to membrane damage. This Pickering emulsion with membrane-lytic ability represents a potential self-anticancer nanocarrier.

Deciphering cell–cell interactions and communication in the tumor microenvironment and unraveling intratumoral genetic heterogeneity via single-cell genomic sequencing

ABSTRACT A tumor’s heterogeneity has important implications in terms of its clonal origin, progression, stemness, and drug resistance. Therefore, because of its significance in treatment, it is important to understand the gene expression pattern of a single cell, track gene expression or mutation in heterogeneous cells, evaluate the clonal origin of cancer cells, and determine the selective evolution of different subpopulations of cancer cells. Researchers are able to trace a cell’s mutation and identify different types of tumor cells by measuring the whole transcriptome with single-cell sequencing (scRNA-seq). This technology provides a better understanding of the molecular mechanisms driving tumor growth than that offered by traditional RNA sequencing methods. In addition, it has revealed changes in the mutations and functions of somatic cells as a tumor evolves; it has also clarified immune cell infiltration and activation. Research on scRNA-seq technology has recently advanced significantly, suggesting new strategies for the treatment of cancer. In short, cancer researchers have become increasingly dependent on scRNA-seq. This paper reviews the development, detection principles, and processes of scRNA-seq technology and their application in tumor research. It also considers potential clinical applications.

Bench to Bedside: New Therapeutic Approaches with Extracellular Vesicles and Engineered Biomaterials for Targeting Therapeutic Resistance of Cancer Stem Cells.

Cancer has recently been the second leading cause of death worldwide, trailing only cardiovascular disease. Cancer stem cells (CSCs), represented as tumor-initiating cells (TICs), are mainly liable for chemoresistance and disease relapse due to their self-renewal capability and differentiating capacity into different types of tumor cells. The intricate molecular mechanism is necessary to elucidate CSC's chemoresistance properties and cancer recurrence. Establishing efficient strategies for CSC maintenance and enrichment is essential to elucidate the mechanisms and properties of CSCs and CSC-related therapeutic measures. Current approaches are insufficient to mimic the in vivo chemical and physical conditions for the maintenance and growth of CSC and yield unreliable research results. Biomaterials are now widely used for simulating the bone marrow microenvironment. Biomaterial-based three-dimensional (3D) approaches for the enrichment of CSC provide an excellent promise for future drug discovery and elucidation of molecular mechanisms. In the future, the biomaterial-based model will contribute to a more operative and predictive CSC model for cancer therapy. Design strategies for materials, physicochemical cues, and morphology will offer a new direction for future modification and new methods for studying the CSC microenvironment and its chemoresistance property. This review highlights the critical roles of the microenvironmental cues that regulate CSC function and endow them with drug resistance properties. This review also explores the latest advancement and challenges in biomaterial-based scaffold structure for therapeutic approaches against CSC chemoresistance. Since the recent entry of extracellular vesicles (EVs), cell-derived nanostructures, have opened new avenues of investigation into this field, which, together with other more conventionally studied signaling pathways, play an important role in cell-to-cell communication. Thus, this review further explores the subject of EVs in-depth. This review also discusses possible future biomaterial and biomaterial-EV-based models that could be used to study the tumor microenvironment (TME) and will provide possible therapeutic approaches. Finally, this review concludes with potential perspectives and conclusions in this area.

Highly sensitive monitoring of telomerase activity in living cells based on rapidly triggered cascade amplification reaction

Telomerase is an important potential biomarker for the study of tumor progression. Herein, we designed a cascade-amplification-reaction-based nanoprobe for intracellular telomerase detection based on the integration of rolling circle amplification (RCA) and catalytic hairpin assembly (CHA) onto MnO2 nanosheets. Firstly, MnO2 nanosheets rapidly delivered and released signal amplification units into cells, and very short telomerase extension products formed RCA circular templates and initiated the exponential RCA, producing enriched telomere sequence amplification products. Then the amplification products specifically triggered the CHA process and numerous H1/H2 complexes were formed, realizing the exponential amplification of fluorescence signals. The detection limit is as low as 1 LoVo cell for telomerase activity in cell extract. We further designed a microfluidic chip with six independent cell culture regions for in situ fluorescence imaging. Simultaneous detection of six types of cells was realized on the chip, and only 1–2 μL of cell suspension and reagents are needed. Our detection method features faster response speed and stronger fluorescence signal. Telomerase in living cells showed strong fluorescence signal within 1.5 h, and tumor cells were effectively distinguished from normal cells. Telomerase activities of different types of tumor cells and activity changes were both monitored conveniently. These results demonstrate that this method holds the potential for the sensitive detection of low abundance biomarkers in living cells, and will contribute to cancer diagnosis, cancer treatment and telomerase-related drug screening.

Measurement of the Compressibility of Cell and Nucleus Based on Acoustofluidic Microdevice.

Cell mechanics play an important role in tumor metastasis, malignant transformation of cells, and radiosensitivity. During these processes, studying the mechanical properties of the cells is often challenging. Conventional measurement methods based on contact such as compression or stretching are prone to cause cell damage, affecting measurement accuracy and subsequent cell culture. Measurements in adherent state can also affect accuracy, especially after irradiation since ionizing radiation will flatten cells and enhance adhesion. Here, a cell mechanics measurement system based on acoustofluidic method has been developed. The cell compressibility can be obtained by recording the cell motion trajectory under the action of the acoustic force, which can realize fast and non-destructive measurement in suspended state. This paper reports in detail the protocols for chip design, sample preparation, trajectory recording, parameter extraction and analysis. The compressibility of different types of tumor cells was measured based on this method. Measurement of the compressibility of nucleus was also achieved by adjusting the resonance frequency of the piezoelectric ceramic and the width of the microchannel. Combined with the molecular level verification of immunofluorescence experiments, the cell compressibility before and after drug-induced epithelial to mesenchymal transition (EMT) were compared. Further, the change of cell compressibility after X-ray irradiation with different doses was revealed. The cell mechanics measurement method proposed in this paper is universal and flexible and has broad application prospects in scientific research and clinical practice.

Pulsatilla chinensis (Bge.) Regel: A Systematic Review on Anticancer of Its Pharmacological Properties, Clinical Researches and Pharmacokinetic Studies.

Pulsatilla chinensis (Bge.) Regel (PC) is one of the most commonly used Chinese medicines and has a history of thousands of years. This article reviews the research results of anti-cancer activity and its mechanism of action obtained from experimental, clinical, pharmacokinetic and bioinformatic studies in recent years. A large number of studies have shown that PC exerts had anti-cancer effects on different types of tumor cells by inhibiting cell proliferation, inducing apoptosis, inhibiting cell cycle and energy metabolism, inducing autophagy, and inhibiting angiogenesis. The literature has shown that PC can trigger the expression of autophagy-related molecules, activate the mitochondrial apoptotic pathway, inhibit the phosphorylation of PI3K downstream factors, down-regulate the expression of glycolysis-related proteins, and regulate a series of cancer-related signal pathways and proteins. The molecular mechanisms involved in PC include signal pathways such as Notch, PI3K/AKT/m TOR, AKT/mTOR, and MEK/ERK. The article also discusses the derivatives of the active ingredients in PC, which greatly improved the anti-cancer effect. In conclusion, this review provides a comprehensive overview of the biological effects and mechanisms of PC against cancer. The analysis of the literature shows that PC can be used as a potential drug candidate for the treatment of cancer.

Abstract 3531: Sensitizing immune unresponsive colorectal cancers to immune checkpoint inhibitors through MAVS overexpression

Abstract Background: The majority of colorectal carcinomas (CRCs) are insensitive to anti-PD-1/PD-L1 immune checkpoint inhibitor (ICI) antibodies. While there are many causes for ICI insensitivity, recent studies suggest that the suppression of innate immunity through a loss of innate immune adaptor expression could be a root cause of this insensitivity and an important factor in the evolution of tumor immunosuppression. Methods: We interrogated mitochondrial antiviral signaling gene (MAVS) expression through bioinformatics analyses of multiple datasets to validate its suppression in clinical samples. We then engineered MAVS expressing tumor cells and tested its ability to elicit innate and adaptive anti-tumor immunity using both in vitro and in vivo approaches, which we then confirmed using MAVS expressing viral vectors. Finally, we observed that MAVS stimulated PD-L1expression in different types of tumor cells and then assessed the combination of PD-L1 ICI antibodies with MAVS tumor expression in vivo. Results: MAVS was significantly downregulated in CRCs, but its re-expression could stimulate broad cellular interferon-related responses, in both murine and patient-derived CRCs. In vivo, local MAVS expression elicited significant anti-tumor responses in both immune-sensitive and insensitive CRC models, through the stimulation of an interferon responsive axis that elicited tumor antigen-specific adaptive immunity. Critically, we found that tumor-intrinsic MAVS expression triggered systemic adaptive immune responses that enabled abscopal CD8+ T cell cytotoxicity against distant CRCs. As MAVS also induced PD-L1 expression, we further found synergistic anti-tumor responses in combination with anti-PD-L1 ICIs. Conclusion: These data demonstrate that intratumoral MAVS expression results in local and systemic tumor antigen-specific T cell responses, which could be combined with PD-L1 ICI to permit effective anti-tumor immunotherapy in ICI resistant cancers. We are further exploring to overcome the ICI insensitivity of other types of solid tumor, with MAVS-delivery strategies other than viral vectors. Citation Format: Bin-Jin Hwang, Li-Chung Tsao, Chaitanya Acharya, Timothy Trotter, Pankaj Agarwal, Tao Wang, Junping Wei, Xiao-Yi Yang, Gang-jun Lei, Takuya Osada, Herbert Kim Lyerly, Michael A. Morse, Zachary Hartman. Sensitizing immune unresponsive colorectal cancers to immune checkpoint inhibitors through MAVS overexpression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3531.