Bornyl acetate was investigated for its anticancer activity against the human gastric cancer cell line SGC-7901, both alone and in combination with 5-fluorouracil (5-FU).
In MTT assays, bornyl acetate treatment for 48 hours induced a dose-dependent growth inhibitory effect on SGC-7901 cells at concentrations of 1.5, 3, 6, 12, 24, 48, and 96 µM. Based on these results, a concentration of 48 µM was selected for combination studies.
The combination of bornyl acetate (48 µM) and 5-FU (1.5 µM) for 48 hours resulted in significantly greater inhibition of cancer cell proliferation compared to treatment with either compound alone, indicating a synergistic enhancement of anticancer activity.
In a colony formation assay, the combination of bornyl acetate (48 µM) and 5-FU (1.5 µM) for 48 hours decreased the number of SGC-7901 cell colonies more effectively than individual treatments.
Phase contrast microscopy revealed that the combination treatment induced more pronounced morphological changes, including cell shrinkage and a reduction in cell population, compared to individual treatments.
Fluorescence microscopy with Hoechst 33342 and acridine orange staining showed that the combination treatment for 48 hours induced characteristic apoptotic features, such as chromatin condensation, nuclear fragmentation, and nuclear shrinkage, to a greater extent than either agent alone. Acridine orange staining showed an increased number of orange-stained apoptotic cells after combination treatment.
DNA fragmentation analysis by agarose gel electrophoresis demonstrated that the combination of bornyl acetate (48 µM) and 5-FU (1.5 µM) for 48 hours resulted in a more obvious DNA ladder pattern compared to individual treatments. This effect was also time-dependent, with increasing fragmentation observed over time.
Flow cytometric analysis with Annexin V-FITC/PI staining showed that treatment with bornyl acetate (48 µM) for 48 hours increased the percentage of apoptotic cells (early + late) to 15.4%, compared to 4.4% in the control. The combination treatment with bornyl acetate (48 µM) and 5-FU (1.5 µM) for 48 hours further increased the apoptotic cell percentage to 55.9%, which was higher than 5-FU alone (39%).
Cell cycle analysis by flow cytometry with PI staining revealed that treatment with bornyl acetate (48 µM) for 48 hours, and more potently its combination with 5-FU (1.5 µM), induced a significant increase in the percentage of cells in the G2/M phase, indicating G2/M cell cycle arrest. [2]

Given that it has one of the highest FD factors, bornyl acetate is one of the most significant odorants in fresh ginger juice. Additionally, it's thought that bornyl acetate is a key sensory component of fresh Japanese ginger's scent [1]. In vitro, bornyl acetate (0-96 μM) inhibits the development of human gastric cancer cells in a dose-dependent manner [2]. Effect of 5-FU and bornyl acetate (48 μM) combined on the apoptotic death of human gastric cancer SGC-7901 cells [2].

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Cell Culture: Human gastric cancer SGC-7901 cells were cultured in RPMI-1640 medium supplemented with 10% fetal calf serum, 100 IU/ml penicillin, and 100 µg/ml streptomycin at 37°C in a 5% CO₂ incubator.
MTT Assay for Cell Viability: Cells were seeded in 96-well plates and treated with various concentrations of bornyl acetate (0, 1.5, 3, 6, 12, 24, 48, 96 µM), 1.5 µM 5-FU, or their combination for 48 hours. MTT solution was added, and after 2 hours of incubation, formazan crystals were dissolved in DMSO. Optical density was measured at 570 nm, and cell viability was calculated as a percentage of the control.
Colony Formation Assay: Cells were suspended in RPMI-1640 with 0.5% agar and seeded onto a base layer of 0.8% agar in 6-well plates. They were treated with bornyl acetate (48 µM), 5-FU (1.5 µM), or their combination for 48 hours and incubated for 1 week. Colonies were fixed, stained with crystal violet, and counted.
Fluorescence Microscopy for Apoptosis: Cells grown on coverslips were treated for 48 hours. They were then stained with Hoechst 33342 and acridine orange for 1 hour. Morphological changes indicative of apoptosis (chromatin condensation, nuclear fragmentation) were observed under a fluorescence microscope.
DNA Fragmentation Assay: Treated cells were collected and lysed. The lysate was treated with RNase A and proteinase K. DNA was extracted using phenol/chloroform/isoamyl alcohol, precipitated with ethanol, and dissolved in TE buffer. DNA samples were subjected to electrophoresis on a 2% agarose gel to visualize DNA laddering.
Flow Cytometry for Apoptosis (Annexin V/PI): Treated cells were harvested, washed, and resuspended in binding buffer. They were stained with FITC-conjugated Annexin V and PI for 30 minutes in the dark. Samples were analyzed by flow cytometry to quantify early (Annexin V+/PI-) and late (Annexin V+/PI+) apoptotic cells.
Flow Cytometry for Cell Cycle Analysis: Treated cells were harvested, fixed in ice-cold 70% ethanol, and stored at -20°C. Before analysis, cells were washed and resuspended in PBS containing PI and RNase A. DNA content was measured by flow cytometry to determine the percentage of cells in G0/G1, S, and G2/M phases. [2]

[1]. Identification of Ginger (Zingiber officinale Roscoe) Volatiles and Localization of Aroma-Active Constituents by GC-Olfactometry. J Agric Food Chem. 2017 May 24;65(20):4140-4145.

[2]. Synergistic enhancement of the antitumor activity of 5-fuorouracil by bornyl acetate in SGC-7901 human gastric cancer cells and the determination of the underlying mechanism of action. J BUON. Jan-Feb 2016;21(1):108-17.

Borneol acetate has been reported in Magnolia officinalis, Perilla frutescens, and other organisms with relevant data. See also: Myrrh (partial).

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Bornyl acetate is a monoterpenoid natural product found in the essential oils of many plant species. This study investigates its potential to enhance the anticancer activity of the conventional chemotherapeutic drug 5-fluorouracil (5-FU) in human gastric cancer cells. The rationale is to find natural compounds that can be combined with existing drugs to increase therapeutic efficacy and potentially reduce side effects by allowing the use of lower doses of the more toxic chemotherapeutic agent. The research demonstrates that bornyl acetate synergistically enhances the anti-proliferative and pro-apoptotic effects of 5-FU in SGC-7901 cells. The combination treatment induces more potent G2/M cell cycle arrest and DNA fragmentation. These findings suggest that bornyl acetate could be a promising candidate for combination therapy in gastric cancer treatment, warranting further in vivo and mechanistic studies. [2]

C12H20O2

196.29

196.146

76-49-3

(-)-Bornyl acetate;5655-61-8;(+)-Bornyl acetate;20347-65-3;Isobornyl acetate;125-12-2

93009

Colorless to light yellow liquid

1.0±0.1 g/cm3

223.5±0.0 °C at 760 mmHg

27ºC

87.4±6.0 °C

0.1±0.4 mmHg at 25°C

1.480

3.6

0

2

2

14

270

3

CC1(C)[C@@]2(C)[C@H](OC(C)=O)C[C@]1([H])CC2

KGEKLUUHTZCSIP-HOSYDEDBSA-N

InChI=1S/C12H20O2/c1-8(13)14-10-7-9-5-6-12(10,4)11(9,2)3/h9-10H,5-7H2,1-4H3/t9-,10+,12+/m0/s1

[(1S,2R,4S)-1,7,7-trimethyl-2-bicyclo[2.2.1]heptanyl] acetate

Bornyl acetate NSC-407158 NSC 407158

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

DMSO : ≥ 83.33 mg/mL (~424.52 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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