Dehydroeffusol (DHE) activates the stress-responsive MEKK4-MKK3-p38 MAPK signaling axis and induces the expression of the tumor-suppressive endoplasmic reticulum (ER) stress gene DDIT3 (CHOP/GADD153).
Concurrently, it inhibits the tumor-adaptive ER stress pathway by downregulating ATF6 and its downstream target GRP78, and suppresses the ERK signaling pathway. [1]

The expression of VE-cadherin, a crucial gene for angiogenesis mimicry, is inhibited by dehydroeffusol [2]. MMP2 expression and activity are decreased in gastric cancer cells by dehydroeffusol [2].

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Dehydroeffusol inhibited the proliferation of 14 human cancer cell lines, including gastric, colon, melanoma, and ovarian cancers, with IC50 values ranging from 11.24 µM to 93.68 µM. The human gastric cancer cell line MGC803 was the most sensitive, with an IC50 of 11.24 µM. The immortalized normal human gastric mucosal cell line GES-1 was less sensitive (IC50 = 42.0 µM). [1]
DHE (8-24 µM) significantly reduced the colony-forming ability of MGC803 cells in soft agar assays in a dose-dependent manner. [1]
DHE treatment (16-24 µM) induced G0/G1 phase cell cycle arrest in MGC803 cells. [1]
DHE induced moderate apoptosis in MGC803 cells, as shown by Annexin V/PI staining (26.0% at 16 µM and 48.9% at 24 µM after 48h), electron microscopy revealing apoptotic morphology, decreased BCL2 protein, and increased cleaved caspase-3 and cleaved PARP levels. This apoptosis was partially caspase-dependent, as a pan-caspase inhibitor reduced apoptosis by ~24.3%. [1]
DHE (8-24 µM) markedly induced ER stress in MGC803 cells, as detected by ER-Tracker Red staining (increased 1.96 to 5.42-fold), and caused mitochondrial dysfunction indicated by decreased mitochondrial membrane potential (JC-1 staining). [1]
At the molecular level, DHE dose-dependently increased mRNA and protein levels of the tumor-suppressive ER stress markers ATF4 and DDIT3. It concurrently decreased mRNA and protein levels of the tumor-adaptive ER stress markers ATF6 (inactive form) and GRP78. [1]
DHE enhanced the transcriptional activity of the DDIT3 gene promoter. [1]
DHE selectively activated the MEKK4-MKK3-p38 MAPK pathway, increasing phosphorylation of MEKK4, MKK3, and p38. It did not activate JNK but significantly inhibited ERK phosphorylation. The p38 inhibitor SB203580 attenuated DHE-induced DDIT3 upregulation and apoptosis. [1]

In a subcutaneous xenograft model using SCID mice implanted with MGC803 gastric cancer cells, daily intraperitoneal administration of Dehydroeffusol (60 mg/kg/day) for 34 days significantly inhibited tumor growth. The average tumor weight in the DHE-treated group was 360.0 ± 71.3 mg compared to 801.7 ± 211.4 mg in the control group, representing a 55.1% inhibition rate. No significant differences in mouse body weight or organ indices (heart, liver, spleen, lung, kidney) were observed between the treated and control groups. [1]

Cell Growth Inhibition Assay: Cells were seeded in 96-well plates and treated with various concentrations of DHE. Cell growth was assessed using a resazurin-based viability assay. The IC50 value was calculated using appropriate software. [1]
Colony Formation Assay: Cells were treated with DHE for 24 hours, harvested, and counted. A base layer of agarose-medium mixture was prepared in a dish. Treated cells were suspended in a medium-low melting agarose mixture and plated on top of the base layer. After solidification, dishes were incubated for 14 days, and colonies were counted under a microscope. [1]
Cell Cycle Analysis: Cells treated with DHE were harvested, fixed, stained with propidium iodide (PI), and analyzed by flow cytometry to determine the distribution of cells in different cell cycle phases. [1]
Apoptosis Assay: Cells treated with DHE were harvested, stained with Annexin V-FITC and PI, and analyzed by flow cytometry to quantify apoptotic and necrotic cell populations. [1]
Western Blot Analysis: Proteins were extracted from treated cells using a mammalian protein extraction reagent. Equal amounts of protein were separated by SDS-PAGE, transferred to membranes, blocked, and incubated with specific primary antibodies overnight. After washing, membranes were incubated with HRP-conjugated secondary antibodies, and protein bands were visualized using chemiluminescence detection reagents. [1]
Reverse Transcription-PCR (RT-PCR): Total RNA was extracted from treated cells. cDNA was synthesized using reverse transcriptase and oligo(dT) primers. PCR amplification was performed using gene-specific primers for DDIT3, GRP78, ATF4, and ATF6. PCR products were analyzed by agarose gel electrophoresis. [1]
ER Stress and Mitochondrial Membrane Potential Assay: For ER stress, treated cells were stained with an ER-specific fluorescent tracker dye and analyzed by flow cytometry. For mitochondrial membrane potential, treated cells grown on coverslips were stained with JC-1 dye and observed under a fluorescence microscope; a shift from red to green fluorescence indicates loss of mitochondrial membrane potential. [1]
Promoter-Luciferase Reporter Assay: Promoter fragments of the DDIT3 gene were cloned into a luciferase reporter vector. Cells were stably transfected with these constructs. After treatment with DHE, cell lysates were prepared, and luciferase activity was measured using a dual-luciferase reporter assay system. [1]

Xenograft Tumor Model: Female SCID mice (6 weeks old) were subcutaneously injected in the back with 1x10^6 MGC803 human gastric cancer cells. When tumors became palpable, mice were randomly divided into groups. The treatment group received daily intraperitoneal injections of Dehydroeffusol dissolved in a suitable vehicle (60 mg/kg body weight per day). The control group received vehicle only. Injections continued for 34 days. Tumor dimensions were measured regularly, and volume was calculated. At the end of the experiment, mice were sacrificed, tumors and major organs were excised and weighed. [1]

In an in vivo xenotransplantation study, daily intraperitoneal injection of dehydroepiandrosterone (60 mg/kg/day for 34 days) did not result in a significant decrease in mouse body weight, with no significant difference compared to the control group. Furthermore, there were no significant differences in organ indices (relative organ weight) of the heart, liver, spleen, lungs, and kidneys between the treatment and control groups, indicating that no significant organ toxicity was observed at this dose and duration. [1] The study also noted that previous reports have shown that dehydroepiandrosterone has extremely low toxicity in inhibiting angiogenesis mimicry. [1]

[1]. Dehydroeffusol inhibits gastric cancer cell growth and tumorigenicity by selectively inducing tumor-suppressive endoplasmic reticulum stress and a moderate apoptosis. Biochem Pharmacol. 2016 Mar 15;104:8-18.

[2]. Dehydroeffusol effectively inhibits human gastric cancer cell-mediated vasculogenic mimicry with low toxicity. Toxicol Appl Pharmacol. 2015 Sep 1;287(2):98-110.

It has been reported that 5-vinyl-1-methylphenanthrene-2,7-diol exists in Juncus setchuensis, Juncus effusus, and Juncus acutus, and there are related data reports. Dehydrojuncus effusus alcohol (DHE) is a small phenanthrene compound (2,7-dihydroxy-1-methyl-5-vinylphenanthrene, molecular weight = 250) isolated from the traditional Chinese medicine Juncus effusus L. [1] Prior to this study, it has been reported that DHE has anxiolytic, sedative, cytoprotective, anticoagulant, and antiangiogenic activities. This study is the first to report its anti-gastric cancer properties. [1]
The proposed mechanism involves selectively inducing tumor-suppressive endoplasmic reticulum stress responses (via ATF4/DDIT3 upregulation and p38 activation) while inhibiting tumor-adaptive endoplasmic reticulum stress (via ATF6/GRP78 downregulation) and the pro-survival ERK signaling pathway, thereby leading to cell cycle arrest, moderate apoptosis, and inhibition of tumorigenesis. [1]
The core structure of DHE contains two free hydroxyl groups and one vinyl group, which are considered to be modifiable for potential derivative synthesis. [1]

C17H14O2

250.2919

250.099

137319-34-7

137319-34-7

5316810

White to off-white solid

4.355

2

2

1

19

340

0

O([H])C1C([H])=C([H])C2=C(C([H])=C([H])C3=C([H])C(=C([H])C(C([H])=C([H])[H])=C23)O[H])C=1C([H])([H])[H]

GSSPKCPIRDPBQE-UHFFFAOYSA-N

InChI=1S/C17H14O2/c1-3-11-8-13(18)9-12-4-5-14-10(2)16(19)7-6-15(14)17(11)12/h3-9,18-19H,1H2,2H3

5-ethenyl-1-methylphenanthrene-2,7-diol

Dehydroeffusol

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ~100 mg/mL (~399.5 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (9.99 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (9.99 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (9.99 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)