How U-87 MG facilitates brain tumor research?
In 1996, U-87 MG cell line was isolated from the malignant glioma of a 44-year-old female patient. This human primary glioblastoma cell line has epithelial morphology and can produce malignant tumors consistent with glioblastoma in nude mice. It is the most common brain tumor research model. In addition to being commonly used in brain cancer research, U-87 MG cell line is also widely used in biomedical industry, such as pathophysiology research, antibiotic screening, cancer drug research, etc. Ubigene provides wild-type U-87 MG cell line with low passage and good viability. STR authentication report is also provided. It is suitable for the construction of various gene-editing cell models.
Applications of CRISPR/Cas9 technology in U-87 MG cell line
CRISPR/Cas9 technology, as a popular gene-editing technology at present, can be used for the precise construction of various disease models, so as to facilitate research of cancer treatment, pathology, physiology and high-throughput drug screening. This technology has been well applied to U-87 MG cell line.
Case Study 1: Gene knockout U-87 MG help to study on growth inhibition of brain tumor cells
Glioblastoma multiforme (GBM), take up 70% of all gliomas, is the most common brain tumor in adults, but there is no curative treatment at present. All patients will basically relapse even after the maximum surgical resection, radiotherapy and chemotherapy. And the median survival time is only about 15 months. Therefore, we should deeply understand the molecular mechanism of glioma occurrence and development. And looking for new targets of treatment has become one of the research hotspots in recent years.
Some studies have found that the deletion of ACSVL3 in U-87 MG will reduce its tumorigenic properties and affect signal transduction through receptor tyrosine kinase. Therefore, Kolar et al knocked out ACSVL3 from U-87 MG cell line, and conducted multiple analyses to explore the effects of ACSVL3 deletion on the carcinogenicity of U-87 MG cell line and GBM cells. Proteomic analysis showed that the content of enzymes involved in ceramide synthesis was low, the level of TCA enzyme was low and the level of glycolytic enzyme was high in ACSVL3 KO U-87 MG cell line. It was found that the knockout of ACSVL3 would affect the synthesis of lipid raft and ganglioside by fluorescence microscope and thin-layer chromatography (TLC). At the same time, the antibody detecting the mitochondrial outer membrane marker Tom20 was used for immunofluorescence. It was observed that the mitochondrial morphology of ACSVL3 KO U-87 MG was different from that of wild type. In conclusion, Kolar et al. believe that ACSVL3 contributes to the GBM cell growth and proliferation, the synthesis of signal sphingolipids, and this enzyme may contribute to the carbohydrate metabolism involved in mitochondria . This conclusion has certain research significance on how to inhibit the occurrence and expansion of GBM.
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Case study 2: U-87 MG point mutant cell line contributes to the pathophysiology of Gaucher's disease (GD)
Gaucher's disease (GD) is a disease caused by acid β-Autosomal recessive lysosomal deposition caused by glucosidase gene (GBA1) mutation. Pavan et al. Used CRISPR/Cas9 to edit GBA1 in human monocyte THP-1 cell line and glioblastoma U-87 MG cell line to construct isogenic GD model (GBA1 mutant). It was found that the two edited cell lines showed low levels of mutant acidity β-Glucosidase expression, less than 1% residual activity, and a large amount of substrate accumulation. In addition, GBA1 mutant U87 MG showed that the mutant enzyme was retained in the endoplasmic reticulum and degraded by proteasome, triggering the unfolded protein reaction (UPR). Compared with wild-type cells, the accumulation of α-syn in GBA1 mutant U87 MG could increase interleukin-1β, and the mutant cell line has higher cell mortality . These results provide a theoretical basis for the study of the pathophysiological mechanism of GD.
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Case study 3: Multiple U-87 MG point mutation cell lines contribute to cancer therapy development
Antibody drug conjugates can target cells and genes with high specificity. Kwon et al constructed multiple variants of TP53 gene in U-87 MG using CRISPR/Cas9 system, and determined that the main transcriptional difference was due to the loss of p53 function. Using transcriptome data, they predicted which mutant clones had less phenotypic difference from the wild-type, so as to screen the best candidate genes that can be used as a drug delivery test platform. Further experiments on cell morphology, proliferation rate and target antigen mediated uptake also confirmed their hypothesis. According to the results of comprehensive analysis, they had successfully screened the most suitable mutant clone. This study provides a great idea for exploring cancer therapeutic reagents .
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 Kolar, Elizabeth Anne. Effects of ACSVL3 Knockout on Lipid and Glucose Metabolism in Malignant Glioma Cells. Diss. Johns Hopkins University, 2016.
 Pavan, Eleonora, et al. "CRISPR/Cas9 editing for Gaucher disease modelling." International journal of molecular sciences 21.9 (2020): 3268.
 Kwon, Andrew Tae-Jun, et al. "Development of p53 knockout U-87 MG cell line for unbiased drug delivery testing system using CRISPR-Cas9 and transcriptomic analysis." Journal of Biotechnology 332 (2021): 72-82.