Scalable Formation of Highly Viable and Functional Hepatocellular Carcinoma Spheroids in an Oxygen-Permeable Microwell Device for Anti-Tumor Drug Evaluation.

For high-throughput anti-cancer drug screening, microwell arrays may serve as an effective tool to generate uniform and scalable tumor spheroids. However, microwell arrays are commonly anchored in non-oxygen-permeable culture plates, leading to limited oxygen supply for avascular spheroids. Herein, a polydimethylsiloxane (PDMS)-based oxygen-permeable microwell device is introduced for generating highly viable and functional hepatocellular carcinoma (HCC) spheroids. The PDMS sheets at the bottom of the microwell device provide a high flux of oxygen like in vivo neighboring hepatic sinusoids. Owing to the better oxygen supply, the generated HepG2 spheroids are larger in size and exhibit higher viability and proliferation with less cell apoptosis and necrosis. These spheroids also exhibit lower levels of anaerobic cellular respiration and express higher levels of liver-related functions. In anti-cancer drug testing, spheroids cultured in PDMS plates show a significantly stronger resistance against doxorubicin because of the stronger stem-cell and multidrug resistance phenotype. Moreover, higher expression of vascular endothelial growth factor-A produces a stronger angiogenesis capability of the spheroids. Overall, compared to the spheroids cultured in conventional non-oxygen-permeable plates, these spheroids can be used as a more favorable model for early-stage HCCs and be applied in high-throughput anti-cancer drug screening.

Establishment and characterization of the NCC-GCTB4-C1 cell line: a novel patient-derived cell line from giant cell tumor of bone.

Giant cell tumor of bone (GCTB) is a rare osteolytic intermediate bone tumor that harbors a pathogenic H3F3A gene mutation and exhibits characteristic histology. The standard curative treatment for GCTB is complete surgical resection, but it frequently results in local recurrence and, more rarely, metastasis. Therefore, effective multidisciplinary treatment is needed. Although patient-derived tumor cell lines are promising tools for preclinical and basic research, there are only four available cell lines for GCTB in public cell banks. Thus, the aim of this study was to establish a novel GCTB cell line. Using surgically resected tumor tissues from a patient with GCTB, we established a cell line named NCC-GCTB4-C1. The cells harbored the typical H3F3A gene mutation and exhibited constant proliferation and invasive capabilities. After characterizing NCC-GCTB4-C1 cell behaviors, we conducted high-throughput screening of 214 anti-tumor drugs and identified seven effective drugs. Comparing the results of high-throughput screening using NCC-GCTB4-C1 cell line with the results using NCC-GCTB1-C1, NCC-GCTB2-C1, and NCC-GCTB3-C1 cell lines that we previously established, four drugs were in common effective. This study showed potential drugs for the treatment of GCTB. These data indicate that NCC-GCTB4-C1 has the potential to be a powerful tool in preclinical and basic research on GCTB.

Repositioning of Hypoglycemic Drug Linagliptin for Cancer Treatment.

BACKGROUND: Drug repositioning, development of new uses for marketed drugs, is an effective way to discover new antitumor compounds. In this study, we used a new method, filtering compounds via molecular docking to find key targets combination. METHODS: The data of gene expression in cancer and normal tissues of colorectal, breast, and liver cancer were obtained from The Cancer Genome Atlas Project (TCGA). The key targets combination was obtained from the protein-protein interaction network (PPI network) and the correlation analysis of the targets. Molecular docking was used to reposition the drugs which were obtained from DrugBank. MTT proliferation assay and animal experiments were used to verify the activity of candidate compounds. Flow cytometric analysis of proliferation, cell cycle and apoptosis, slice analysis, gene regulatory network, and Western blot were performed to elucidate the mechanism of drug action. RESULTS: CDK1 and AURKB were identified as a pair of key targets by the analysis of different expression gene from TCGA. Three compounds, linagliptin, mupirocin, and tobramycin, from 12 computationally predicted compounds, were verified to inhibit cell viability in HCT116 (colorectal), MCF7 (breast), and HepG2 (liver) cancer cells. Linagliptin, a hypoglycemic drug, was proved to inhibit cell proliferation by cell cycle arrest and induce apoptosis in HCT116 cells, and suppress tumor growth in nude mice bearing HCT116 cells. Linagliptin reduced the tumor size and decreased the expression of Ki67, a nuclear protein expressed in all proliferative cells. Gene regulatory network and Western blot analysis suggested that linagliptin inhibited tumor cell proliferation and promoted cell apoptosis through suppressing the expression and phosphorylation of Rb, plus down-regulating the expression of Pro-caspase3 and Bcl-2, respectively. CONCLUSION: The combination of key targets based on the protein-protein interaction network that were built by the different gene expression of TCGA data to reposition the marketed drugs turned out to be a new approach to discover new antitumor drugs. Hypoglycemic drug linagliptin could potentially lead to novel therapeutics for the treatment of tumors, especially for colorectal cancer. Gene regulatory network is a valuable method for predicting and explaining the mechanism of drugs action.

Facile and ultrasensitive fluorescence sensor platform for tumor invasive biomaker ß-glucuronidase detection and inhibitor evaluation with carbon quantum dots based on inner-filter effect.

Early detection and diagnosis have great practical significances for the effective prevention and treatment of cancer. In this study, we developed a novel, facile and ultra-sensitive fluorescence assay for the determination of tumor invasive biomarker ß-glucuronidase (GLU) based on the inner-filter effect (IFE). The nitrogen-doped carbon quantum dots (N-CQDs) with green photoluminescence were employed as the fluorophore in IFE, and 4-nitrophenyl-ß-D-glucuronide (PNPG) was used to act as GLU substrate, and GLU catalytic product (p-nitrophenol (PNP)) was capable of acting as the robust absorber in IFE to turn off the fluorescence of N-CQDs due to the complementary overlap between the absorption of PNP and the excitation of N-CQDs. Thus, signal of GLU activity could be recorded by the fluorescence intensity of N-CQDs. Unlike other fluorescence sensing mechanism such as fluorescence resonance energy transfer (FRET) or photoinduced electron transfer (PET), IFE has no requirement for electron or energy transfer process or any chemical modification of fluorophore, which makes our assay more flexible and simple. The proposed method exhibited a good linear relationship from 1UL(-1) to 60UL(-1) (R(2)=0.9967) with a low detection limit of 0.3UL(-1). This method was also successfully applied to the analysis of serum samples and the inhibitor screening from natural product. The developed sensor platform was proven to be reliable, facile, sensitive, and selective, making it promising as a candidate for GLU activity detection in clinic tumor diagnose and anti-tumor drug screening.