KYSE-150 cells are poorly differentiated esophageal adenocarcinoma cells that were isolated from the neck esophagus of a 49-year-old Japanese female patient who had received radiotherapy. When researchers discovered this cell line, the patient's cancer tissue had invaded the adjacent tissues. The KYSE-150 cell line is an epithelial cell with an adherent monolayer. It has been reported that cells carry increased oncogenes c-erb-B (8 times) and cyclin D1 (4 times) and that cells can form tumors in nude mice, so KYSE-150 cell line is often used in cancer research involving gene editings, such as CRISPR gene knockout/knock-in and point mutation.
Applications:
1. EZH2 knockout in KYSE-150 cells altered PSMA3-AS1-induced proliferation and migration in esophageal cancer cells
PSMA3-AS1 expression is up-regulated and positively correlated with tumor size and metastasis in esophageal cancer patients. To further explore the biological roles of PSMA3-AS1 in esophageal cancer cells, researchers established stable CRISPR PSMA3-AS1-overexpressing cell lines via lentiviral infection in KYSE150 and KYSE450 cell lines (which have low PSMA3-AS1 expression) and validated the up-regulation of PSMA3-AS1 by RT-qPCR.
CRISPR/Cas9 gene-editing of EZH2 was successfully performed in KYSE150 and KYSE450cells as confirmed by a significant reduction in EZH2 protein expression. CCK-8 and colony formation assays showed that PSMA3-AS1 overexpression did not affect the proliferation of esophageal cancer cells with CRISPR EZH2 knocked out (Figure 6B and 6C). According to wound healing and transwell migration assays, wounding healing and cell migration were not increased in ESCC EZH2-knock out KYSE-150 cells compared to negative control cells.
Clinical-pathological characteristics illustrated that increased expression of PSMA3-AS1 was positively associated with distant metastasis, larger tumor sizes, and a worse prognosis for ESCC patients.
2. CRISPR Over-expressed KYSE-150 cells increase chemo-resistance in ESCC
Whole-genome clustered regularly interspaced short palindromic repeats/CRISPR associated (CRISPR/Cas)-based a lentiviral library is a powerful tool for genome-scale gain-of-function or loss-of-function screening. This system has been proved to be highly effective in identifying drug-resistant genes in vitro. Shalem, Kurata, and Joung have screened out essential genes for drug resistance in melanoma and AML using the CRISPR knockout library. Some researchers attempted to combine the CRISPR library screening strategy with RNA sequencing technology in KYSE-150 Cells to explore the critical genes and potential mechanism for chemo-resistance in ESCC.
To increase the chance of identifying essential genes involved in PTX resistance, an integrated analysis was performed to combine EN-genes in genome-scale CRISPR screening and differentially expressed genes (DE-genes) in KYSE-150 cell line.
Researchers evaluated the potential of CDKN1A, ELAVL2, and TSPAN4 to promote chemo-resistance in ESCC cells KYSE-150. The results showed that overexpression of CDKN1A, ELAVL2, or TSPAN4 could significantly increase the resistance to PTX in both KYSE-180 and KYSE-150 cells. Moreover, overexpressed CDKN1A, ELAVL2, or TSPAN4 could also contribute to DDP-resistance in KYSE-150 cells.
3. CRISPR/Cas9 mediated knockout of DEPTOR in KYSE-510 cells significantly promoted cellular proliferation, migration, and invasion
Researchers found that the expression of DEPTOR negatively regulates the tumorigenic activities of ESCC cell lines. Furthermore, ectopic DEPTOR expression caused significant suppression of the cellular proliferation, migration, and invasion of KYSE150 cells, which has the lowest expression level of DEPTOR. Meanwhile, CRISPR/Cas9 mediated knockout of DEPTOR in KYSE-510 cells significantly promoted cellular proliferation, migration, and invasion. Besides, in vivo assays further revealed that tumor growth was significantly inhibited in xenografts with ectopic DEPTOR expression as compared to untreated KYSE150 cells, and was markedly enhanced in DEPTOR knockout KYSE-510 cells.
In this study, scientists generated stable cell lines that either overexpressing DEPTOR or genetic ablation of endogenous expression of DEPTOR. Since KYSE-150 expresses the lowest endogenous level of DEPTOR among the three cell lines, they stably overexpressed DEPTOR in KYSE-150 cells (pcDNA3.1-DEPTOR). For the same consideration, KYSE-510 cells that express the highest level of DEPTOR were treated with CRISPR/Cas9 system to knockout of DEPTOR (CRISPR-DEPTOR). After the generation of cell lines, pcDNA3.1-DEPTOR displayed a reduced cell proliferation rate as compared to that of KYSE-150 parental cells and empty vector-transfected cells, while CRISPR-DEPTOR cells proliferated significantly faster than control KYSE-510 cells. Furthermore, pcDNA3.1-DEPTOR cells also showed reduced migration. Thus, these results suggested DEPTOR indeed regulates cell proliferation, migration, and invasion in ESCC cells.
Ubigene Biosciences is co-founded by biological academics and elites from China, the United States, and France. We are located in Guangzhou Science City, which serves as a global center for high technology and innovation. Ubigene Biosciences has 1000㎡ office areas and laboratories, involving genome editing, cell biology technology, and zebrafish research. We provide products and services for plasmids, viruses, cells, and zebrafish. We aim to provide customers with better gene-editing tools for cell or animal research.
Make genome editing easier is the goal of Ubigene. We developed CRISPR-U™ (based on CRISPR/Cas9 technology) which is more efficient than general CRISPR/Cas9 in double-strand breaking, and CRISPR-U™ can greatly improve the efficiency of homologous recombination, easily achieve knockout (KO), point mutation (PM) and knockin (KI) in vitro and in vivo. With CRISPR-U™, Ubigene has successfully edit genes on more than 100 cell lines
Ubigene developed CRISPR-B™ which optimizes the microbial gene-editing vectors and process. The efficiency and accuracy are much higher than traditional methods. CRISPR-B™ can be used in gene editing of bacteria and fungi. Easily achieve microbial gene knockout (KO), point mutation (PM) and knockin (KI).
Ubigene has more than 400 types of primary cells, including epithelial cells, endothelial cells, smooth muscle cells and fibroblasts from different species, such as human, rat, and mouse. We can provide a validation report for each primary cell. Our primary cells have been widely used in many research institutes and pharmaceutical enterprises.
References:
1. Yan-Mei Ji, Xue-Feng Zhou, Jun Zhang, Xiang Zheng, Sheng-Bao Li, Zhi-Qiang Wei, Tao Liu, Dong-Liang Cheng, Ping Liu, Kuncheng Song, Tao Tan, Hua Zhu, Jia-Long Guo. DEPTOR suppresses the progression of esophageal squamous cell carcinoma and predicts poor prognosis. 2016 Mar 22; 7(12): 14188–14198. Published online 2016 Feb 16.
2. Qiu BQ, Lin XH, Ye XD, et al. Long non-coding RNA PSMA3-AS1 promotes malignant phenotypes of esophageal cancer by modulating the miR-101/EZH2 axis as a ceRNA. Aging (Albany NY). 2020;12(2):1843‐1856.
3. Zhao, Wen-Si et al. “Genome-scale CRISPR activation screening identifies a role of ELAVL2-CDKN1A axis in paclitaxel resistance in esophageal squamous cell carcinoma.” American journal of cancer research vol. 9,6 1183-1200. 1 Jun. 2019
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