Panoply™ Human MTHFD2 Over-expressing Stable Cell Line

Name: Panoply™ Human MTHFD2 Over-expressing Stable Cell Line
SKU: CSC-SC009879
Availability: InStock

For research use only. Not intended for any clinical use.

Cat. No. : CSC-SC009879

Host Cell : HEK293 (CHO and other cell types are also available) Size : >1x10⁶ frozen cells/vial

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Cell Line Information

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Gene Information

Cat. No.	CSC-SC009879
Description	Using Creative Biogene's proprietary lentiviral vectors, we subclone the target gene into lentivector, generate the lentivirus particles, sequentially infect the cell line HEK293 (other cell types are also available according to your requirements), and select the clones constantly expressing target gene at high level.
Target Gene	MTHFD2
Gene Species	Homo sapiens (Human)
Host Cell	HEK293 (CHO and other cell types are also available)
Host Cell Species	Species varies
Applications	1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Disease research
Size	2 × 10^6 cells / vial
Stability	Validated for at least 10 passages
Quality Control	Negative for bacteria, yeast, fungi and mycoplasma.
Storage	Liquid nitrogen
Shipping	Dry Ice

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.

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

Gene Name	MTHFD2 methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 2, methenyltetrahydrofolate cyclohydrolase [ Homo sapiens ]
Gene Symbol	MTHFD2
Synonyms	NMDMC
Gene Description	methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 2, methenyltetrahydrofolate cyclohydrolase
GeneID	10797
Uni ProtID	P13995
mRNA Refseq	NM_006636.3
Protein Refseq	NP_006627.2
Chromosome Location	2p13.1
Function	magnesium ion binding; methenyltetrahydrofolate cyclohydrolase activity; methylenetetrahydrofolate dehydrogenase (NAD+) activity; methylenetetrahydrofolate dehydrogenase (NADP+) activity; nucleotide binding; phosphate ion binding;
Pathway	C1-unit interconversion, eukaryotes, organism-specific biosystem; C1-unit interconversion, eukaryotes, conserved biosystem; Nucleotide Metabolism, organism-specific biosystem; One Carbon Metabolism, organism-specific biosystem; One carbon pool by folate, organism-specific biosystem; One carbon pool by folate, conserved biosystem; folate transformations I, organism-specific biosystem;
MIM	604887

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MTHFD2, officially known as methylenetetrahydrofolate dehydrogenase (NADP+-dependent) 2, methenyltetrahydrofolate cyclohydrolase, is a nuclear-encoded mitochondrial bifunctional enzyme encoded by the human gene MTHFD2 located on chromosome 2p13.1. This enzyme catalyzes two sequential reactions in the mitochondrial folate pathway: the NADP-dependent conversion of 5,10-methylenetetrahydrofolate to 5,10-methenyltetrahydrofolate, followed by the cyclohydrolase reaction that produces 10-formyltetrahydrofolate. Unlike its cytosolic counterpart MTHFD1, MTHFD2 functions exclusively in the mitochondria and operates as a homodimer with an absolute requirement for magnesium ions and inorganic phosphate. The enzyme plays a central role in one-carbon metabolism, supporting nucleotide biosynthesis, methylation reactions, and redox homeostasis. Under normal physiological conditions, MTHFD2 expression is largely restricted to embryonic and highly proliferative tissues, but it is significantly upregulated in a wide range of human cancers, including breast, lung, colorectal, pancreatic, renal, esophageal, and ovarian carcinomas, where its overexpression is consistently linked to poor clinical prognosis and aggressive tumor behavior.

The Human MTHFD2 Overexpressing Stable Cell Line is a valuable research tool for investigators studying cancer metabolism, mitochondrial biology, and one-carbon metabolic pathways. These cells are engineered to stably express elevated levels of MTHFD2 protein, enabling researchers to model the metabolic reprogramming observed in malignant cells and investigate the mechanistic links between MTHFD2 activity and tumorigenesis. This cell line is particularly well-suited for studies examining how MTHFD2-driven folate metabolism supports cancer cell proliferation, survival under oxidative stress, and resistance to chemotherapeutic agents. Applications include evaluating the effects of MTHFD2 overexpression on cellular redox balance, nucleotide pool maintenance, and mitochondrial function, as well as assessing the impact of metabolic perturbations on downstream signaling pathways such as PI3K/AKT and ERK1/2. Additionally, this stable cell line provides a reliable platform for high-throughput screening of small-molecule inhibitors targeting MTHFD2, making it a useful tool for anticancer drug discovery efforts aimed at exploiting the metabolic vulnerabilities of cancer cells.

Methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) is a bifunctional enzyme localized within the mitochondria. Previous studies have demonstrated that MTHFD2 is highly expressed in various malignant tumors and contributes to cancer initiation and progression. In this study, researchers investigated the impact of MTHFD2 on ovarian cancer progression. The results showed that MTHFD2 is highly expressed in both ovarian cancer tissues and cell lines. Depletion of MTHFD2 inhibited cell proliferation and metastasis. Knockdown of MTHFD2 induced apoptosis and G2/M phase arrest, whereas MTHFD2 overexpression led to an increase in the proportion of cells in the S phase. Mechanistic studies indicated that the inhibitory effects resulting from MTHFD2 knockdown might be associated with the downregulation and reduced activity of the Cyclin B1/Cdc2 complex. Furthermore, MTHFD2 regulates cell growth and invasive capacity by activating STAT3 and the downstream epithelial-mesenchymal transition (EMT) signaling pathway. In summary, MTHFD2 is highly expressed in ovarian cancer and regulates cell proliferation and metastasis, making it a promising therapeutic target.

To investigate the potential molecular mechanisms by which MTHFD2 promotes cell proliferation and invasion, researchers first used the UCSC online database to screen for proteins potentially associated with MTHFD2; the STAT3 signaling pathway-closely linked to cell growth, metastasis, and apoptosis-emerged as a focus of interest. Western blot analysis was employed to measure the levels of p-STAT3, STAT3, and proteins associated with the epithelial-mesenchymal transition (EMT). The results demonstrated that MTHFD2 knockdown inhibited STAT3 phosphorylation in SKOV3 and OVCAR8 cells. Furthermore, the expression levels of N-cadherin and Vimentin-key markers regulating tumor invasiveness-were significantly reduced following MTHFD2 depletion (Figure 1A). To further validate the link between MTHFD2 and the STAT3 signaling pathway, researchers examined MTHFD2 overexpressing SKOV3 and OVCAR8 cells and observed elevated levels of p-STAT3 (Figure 1B). These findings indicate that MTHFD2 promotes ovarian cancer cell progression via the STAT3 signaling pathway (Figure 1C).

Figure 1. MTHFD2 regulates ovarian cancer cell progression via STAT3 pathway. (Li Q, et al., 2021)

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