Human PTGS2 Knockout Cell Line-HeLa
Cat.No. : CSC-RT1517
Host Cell: HeLa Target Gene: PTGS2
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT1517 |
| Cell Line Information | A stable cell line with a homozygous knockout of human PTGS2 using CRISPR/Cas9. |
| Target Gene | PTGS2 |
| Host Cell | HeLa |
| Host Cell Species | Homo sapiens (Human) |
| Shipping | 10^6 cells/tube |
| Storage | Liquid nitrogen |
| Gene ID | 5743 |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Media Type | Cells were cultured in DMEM supplemented with 10% fetal bovine serum. |
| Growth Properties | Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2. Split at 80-90% confluence, approximately 1:4-1:6. |
| Freeze Medium | Complete medium supplemented with 10% (v/v) DMSO |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
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Background
Case Study
Applications
The PTGS2 gene, also known as prostaglandin endoperoxide synthase 2 or cyclooxygenase-2 (COX-2), is a key component in the biosynthesis of prostaglandins such as prostaglandins, prostacyclins, and thromboxanes, which are major regulators of inflammatory processes, pain, fever, and other physiological responses. The PTGS2 gene is located on human chromosome 1 and encodes a particularly inducible enzyme that responds dynamically to a variety of stimuli such as proinflammatory cytokines, growth factors, and bacterial endotoxins. Under normal physiological conditions, PTGS2 expression is relatively low. However, in response to inflammatory signals, it is rapidly upregulated, unlike COX-1, which is constitutively expressed in most tissues and involved in maintaining physiological homeostasis. This inducibility makes COX-2 a key player in acute inflammation and pathophysiological states such as arthritis, cancer, and cardiovascular disease.
The enzyme produced by PTGS2 converts arachidonic acid into prostaglandin H2, which is a precursor for other prostaglandins. Given its important role in inflammation and pain, PTGS2 has become a prime target for nonsteroidal anti-inflammatory drugs (NSAIDs) and selective COX-2 inhibitors. Traditional NSAIDs inhibit both COX-1 and COX-2, which can result in gastrointestinal side effects due to inhibition of COX-1-mediated protective prostaglandins. In contrast, selective COX-2 inhibitors such as celecoxib are designed to reduce inflammation and pain while minimizing adverse gastrointestinal effects by selectively inhibiting the enzyme encoded by PTGS2.
Cytotoxic therapies, in addition to directly inducing cancer cell death, can stimulate immune-dependent tumor growth control or accelerate tumor progression. Here, researchers show that cytotoxic therapies acutely upregulate cyclooxygenase (COX)-2 (also known as Prostaglandin-endoperoxide synthase 2 (PTGS2)) expression and prostaglandin E2 (PGE2) production in cancer cells with pre-existing COX-2 activity. Screening of a compound library of 1280 approved drugs revealed that chemotherapeutic agents from all classes enhance COX-2 transcription while inhibiting cancer cell proliferation. Genetic manipulation of COX-2 expression or its gene promoter region revealed how enhanced COX-2/PGE2 activity after treatment profoundly alters the inflammatory properties of chemotherapy-treated cancer cells in vivo. Pharmacological COX-2 inhibition enhances the efficacy of combined chemotherapy and PD-1 blockade. These findings suggest that COX-2/PGE2 upregulation by dying cancer cells is a major barrier to cytotoxic therapy-driven tumor immunity and reveal a strategy to improve outcomes of combined immunotherapy and chemotherapy treatments.
Here, to assess the contribution of the COX-2/PGE2 axis to the inflammatory phenotype of CTX-treated cells, the researchers compared the effects of injection of cisplatin or 5-FU-treated COX-2 WT and COX-2 (PTGS2) knockout 4T1 cells. Notably, CTX-treated COX-2 knockout cells attracted far fewer neutrophils or monocytes than their COX-2-expressing parental cells and were similar to untreated 4T1 cells (Figure 1), indicating a major contribution of cancer cell-intrinsic COX-2 activity. COX-2 REST cells pre-treated with cisplatin or 5-FU were less effective than COX-2 WT cells in recruiting neutrophils and monocytes and largely phenotypically mimicked CTX-treated COX-2 knockout cells (Figure 1). Together, these results suggest that transcriptional upregulation of COX-2 by cancer cells after CTX, rather than its basal expression, largely explains their ability to stimulate recruitment of inflammatory myeloid cells.
Figure 1. Total number of neutrophils and monocytes recruited by i.p. injection of untreated or CTX-treated 4T1 COX-2 WT, COX-2 knockout (KO), or COX-2 REST cells. (Bell, Charlotte R., et al. 2022)
The applications of the PTGS2 knockout HeLa cell line are vast and can significantly improve our understanding of various biological processes and disease states. Here are some of the key applications:
Cancer Research: PTGS2 is known to be involved in inflammatory processes and is overexpressed in various cancers. The PTGS2 knockout HeLa cell line can be used to study the role of COX-2 in tumor progression, metastasis, and response to therapy.
Drug Screening: This knockout cell line can be used to screen for drugs that specifically target pathways that are dependent on PTGS2. It provides a model to identify compounds that can inhibit or activate alternative pathways, aiding in the development of novel therapeutic agents.
Inflammation and Pain Research: PTGS2 is essential for the synthesis of prostaglandins, which are mediators of inflammation and pain. By knocking out PTGS2, researchers can study the molecular mechanisms behind inflammation and pain, gaining insights that may aid in the development of novel anti-inflammatory and analgesic drugs.
Cell Cycle and Apoptosis: Since PTGS2 plays a role in cell proliferation and survival, PTGS2 knockout HeLa cells can be used to study its involvement in regulating the cell cycle and apoptosis.
Pathway Analysis: PTGS2 is part of the arachidonic acid pathway and is involved in the production of prostaglandins. Using PTGS2 knockout cells, researchers can dissect this pathway to better understand how different enzymes and products interact and lead to physiological and pathological conditions.
For research use only. Not intended for any clinical use.
