Hydroalcoholic Extract of Cuscuta chinensis Suppresses Angiogenesis and Enhances Paclitaxel Activity in the Chick Chorioallantoic Membrane (CAM) Model Cuscuta Anti Angiogenic

Fatemeh Sadat Ahmadi (1), Majid Morovati-Sharifabad (2), Gholam Hossein pourghanbari Marvast (3), Elham Salehi (4), Ehsan Zarei MahmoudAbadi (5)
(1) Department of basic science, Faculty of Veterinary Medicine, Ardakan University, Ardakan, Iran, Iran, Islamic Republic of,
(2) Department of Basic Sciences, Faculty of Veterinary Medicine, Ardakan University, Ardakan, Iran, Iran, Islamic Republic of,
(3) Department of Basic Sciences, Faculty of Veterinary Medicine, Ardakan University, Ardakan, Iran, Iran, Islamic Republic of,
(4) Department of Basic Sciences, Faculty of Veterinary Medicine, Ardakan University, Ardakan, Iran, Iran, Islamic Republic of,
(5) Meybod Genetic Research Center, Meybod, Iran, Iran, Islamic Republic of

Abstract

Background: With the growing resistance of cancers to conventional therapies, there is increasing interest in identifying natural compounds with potent anticancer properties. Cuscuta chinensis, a valuable herb in traditional medicine, has attracted attention for its potential therapeutic effects. Given the critical role of angiogenesis in tumor growth and metastasis, the discovery of effective anti-angiogenic agents is essential. The chick chorioallantoic membrane (CAM) model is a well-established system for studying angiogenesis. This study aimed to evaluate the anti-angiogenic effects of hydroalcoholic extract of C. chinensis on the CAM model.


Methods: This case-control experimental study included six groups of fertilized eggs: control groups and treatment groups receiving paclitaxel (40 µg/kg), C. chinensis extract (100 and 400 mg/ml), and a combination of paclitaxel and extract. Treatments were administered on day 8 of incubation. Angiogenesis was assessed using morphological parameters (number and thickness of major and minor vascular branches measured by ImageJ software) and molecular analysis of VEGF gene expression using real-time PCR. Statistical analysis was performed using SPSS software.


Results: The extract significantly reduced angiogenic parameters in a dose-dependent manner (p < 0.05). Paclitaxel showed stronger inhibition of major vessel formation and enhanced the extract’s effects. Both treatments significantly decreased VEGF expression, with the combination producing greater inhibition.


Conclusion: C. chinensis extract exhibits significant anti-angiogenic activity in the CAM model and may potentiate the effects of paclitaxel.

References

Newman DJ, Cragg GM. Natural products as sources of new drugs over the nearly four decades from 01/1981 to 09/2019. Journal of natural products. 2020;83(3):770-803. doi:10.1021/acs.jnatprod.9b01285.

Cragg GM, Newman DJ. Natural products: a continuing source of novel drug leads. Biochimica et Biophysica Acta (BBA)-General Subjects. 2013;1830(6):3670-95. doi:10.1016/j.bbagen.2013.02.008.

Gangarde P, Kuchekar B, Kuchekar A, Gawade A, Pujari R. Cuscuta reflexa roxb.: A special emphasis on its anticancer, antimicrobial, anti-inflammatory, immunomodulatory potential. Research Journal of Pharmacy and Technology. 2024;17(10):5055-61. doi:10.52711/0974-360X.2024.00777.

Baráth K, Lengyel A, Csiky J. Host specificity of Cuscuta species: is it a cause or a consequence of the habitat preference? Plant and Soil. 2025;513(1):709-21. doi:10.1007/s11104-025-07210-2.

Lozanova V, Teofanova D, Chakarova B, Rusanov K, Pachedjieva K, Tosheva A, et al. The Antioxidant Properties of Extracts of Cuscuta spp. Depend on the Parasite and the Host Species. Antioxidants. 2025;14(7):761. doi:10.3390/antiox14070761.

Baccari H, Bedoui A, Feriani A, Bouallegue A, Sahri N, Khatib S, et al. Protective Effects of Cuscuta australis Against CCl4-Induced Hepatic Injury in Rats: Antioxidant, Anti-Inflammatory, and In Silico Insights. Pharmaceuticals. 2025;18(10):1524. doi:10.3390/ph18101524.

Tursun E, Qiao Y, Dai L, Yu J, Yusuf A, Abudurexiti A. Chemical constituents and biological activities of the whole plant of Cuscuta chinensis Lam. Natural Product Research. 2025:1-6. doi:10.1080/14786419.2025.2487155.

Kim J-H, Seo HJ, Noh BW, He MT, Choi Y-H, Cho EJ, et al. Protective effects of Cuscuta chinensis Lam. extract against learning and memory dysfunction induced by streptozotocin and amyloid β25-35 in vivo model. Archives of Physiology and Biochemistry. 2025;131(5):805-17. doi:10.1080/13813455.2025.2502861.

Seo HJ, Noh BW, He MT, Kim J-H, Noh JS, Cho EJ. Protective Effects of Cuscuta chinensis Lam. Extract against Learning and Memory Dysfunction in Streptozotocin and Amyloid β 25-35-induced in Vivo Model. 한국식품영양과학회 학술대회발표집. 2021:544-. doi:10.1080/13813455.2025.2502861.

Xiao Z, Zheng H, Zhang S, Li Q, Wang Y, Wang H, et al. Unraveling the characteristic structures of Cuscuta chinensis Lam. polysaccharides and its inhibitory effects on non-alcoholic fatty liver disease. International Journal of Biological Macromolecules. 2025:146798. doi:10.1016/j.ijbiomac.2025.146798.

Nitthikan N, Preedalikit W, Supadej K, Chaichit S, Leelapornpisid P, Kiattisin K. Exploring the wound healing potential of a Cuscuta chinensis Extract-Loaded Nanoemulsion-Based Gel. Pharmaceutics. 2024;16(5):573. doi:10.3390/pharmaceutics16050573.

Ara L, Jafari F, Mokaberinejad R. Cuscuta chinensis antidepressant-like effect, the role of 5-HT and the impact on behavioral response in mice. Research Journal of Pharmacognosy. 2022;9(3):1-4. doi:10.22127/rjp.2022.338343.1885.

Tao Y, Chen L, Pan M, Zhu F, Yan J. Tracing anti‐osteoporosis components from raw and salt‐processed semen of Cuscuta chinensis by employing a biochemometrics strategy that integrates ultrasonic‐assisted extraction, quantitation, efficacy assessment in zebrafish, and grey relationship analysis. Journal of separation science. 2021;44(17):3229-36. doi:10.1002/jssc.202100272.

Li Y, Xu Y, Zhang B, Wang Z, Ma L, Sun L, et al. Atractylodes macrocephala Koidz. and Cuscuta chinensis Lam. extract relieves insulin resistance via PI3K/Akt signalling in diabetic Drosophila. Journal of Traditional and Complementary Medicine. 2024;14(4):424-34. doi:10.1016/j.jtcme.2024.01.010.

Zhou L, Liu R, Wang X, Song W, Duan X, Yang Y, et al. Cuscuta chinensis Lam. extracts ameliorate type 2 diabetes by regulating metabolism and gut microbiota. iScience. 2026;29(1). doi:10.1016/j.isci.2025.114474.

Maimaiti A, Xu J, Shi L. An RNA-seq transcriptome analysis for investigating the anti-lung cancer activity of medicinal Cuscuta chinensis Lam plant. British Journal of Nutrition. 2023;130(5):737-49. doi:10.1017/S0007114522003865.

Dermani FK, Saidijam M, Najafi R, Moradkhani S, Mohammadzaheri Z, Beiranvand N, et al. Cytotoxic effects of hydroalcoholic extract of Cuscuta chinensis on PC3 and MCF7 cancer cell lines. Avicenna Journal of Phytomedicine. 2021;11(3):258. doi:10.22038/ajp.2020.16902.

Chen C, Liu Y, Hu J, Gu Y, Li W, Yue H, et al. Herbal Cuscutae Semen Contributes to Oxidative Stress Tolerance and Extends Lifespan via Sirtuin1 in Caenorhabditis elegans. Antioxidants. 2025;14(7):786. doi:10.3390/antiox14070786.

Gao C, Song XD, Chen FH, Wei GL, Guo CY. The protective effect of natural medicines in rheumatoid arthritis via inhibit angiogenesis. Frontiers in Pharmacology. 2024;15:1380098. doi:10.3389/fphar.2024.1380098.

Ai Y, Zhao Z, Wang H, Zhang X, Qin W, Guo Y, et al. Pull the plug: Anti‐angiogenesis potential of natural products in gastrointestinal cancer therapy. Phytotherapy Research. 2022;36(9):3371-93. doi:10.1002/ptr.7492.

Jia S, Li L, Yu C, Peng F. Natural products’ antiangiogenic roles in gynecological cancer. Frontiers in Pharmacology. 2024;15:1353056. doi:10.3389/fphar.2024.1353056.

Wei Q, Zhang Y-h. Flavonoids with anti-angiogenesis function in cancer. Molecules. 2024;29(7):1570. doi:10.3390/molecules29071570.

Shekatkar M, Kheur S, Deshpande S, Sakhare S, Sanap A, Kheur M, et al. Critical appraisal of the chorioallantoic membrane model for studying angiogenesis in preclinical research. Molecular Biology Reports. 2024;51(1):1026. doi:10.1007/s11033-024-09956-x.

Chastel T, Filiberti S, Mitola S, Ronca R, Turtoi A, Corsini M. Protocol for performing angiogenic and tumorigenic assays using the in ovo chick embryo chorioallantoic membrane model. STAR protocols. 2025;6(1). doi:10.1016/j.xpro.2025.103663.

Caplar BD, Togoe MM, Ribatti D, Pop D, Sinescu C, Rominu M, et al. The chick embryo chorioallantoic membrane (CAM) assay: a novel experimental model in dental research. Cureus. 2024;16(11). doi:10.7759/cureus.74714.

Lixiang F, Rongqian Z, Zhang K, Yang W. Comparison of the 2-CT method and the 2-ΔΔCT method for real-time qPCR data analysis. bioRxiv. 2025:2025.07. 16.665089. doi:10.1101/2025.07.16.665089.

Dhiman V, Kumari M, Singh D. Chemoresistance: the hidden barrier in cancer treatment, cancer pathogenesis and therapy. 2025. doi:10.1016/j.cpt.2025.07.001.

Wan Z, Hirche C, Fricke F, Dragu A, Will PA. Chick chorioallantoic membrane as an in vivo model for the study of angiogenesis and lymphangiogenesis. Journal of Vascular Research. 2025;62(2):109-20. doi:10.1159/000542875.

Muraro E, Vinante L, Fratta E, Bearz A, Höfler D, Steffan A, et al. Metronomic chemotherapy: anti-tumor pathways and combination with immune checkpoint inhibitors. Cancers. 2023;15(9):2471. doi:10.3390/cancers15092471.

Wu H-l, Zhou H-x, Chen L-m, Wang S-s. Metronomic chemotherapy in cancer treatment: new wine in an old bottle. Theranostics. 2024;14(9):3548. doi:10.7150/thno.95619.

Al-Sudani BT. Redounding of Cuscuta chinensis Lam. on BxPC-3, HepG2, and U2OS Human Cancer Cell Lines. International Journal of Drug Delivery Technology. 2020;3:354-9. doi:10.25258/ijddt.10.3.8.

Saadoon AS, Alabbassi MG, Mohammed NH. Cuscuta Chinensis potentiate the effect of methotrexate in Rheumatoid Arthritis Induced Rats. Al Mustansiriyah Journal of Pharmaceutical Sciences. 2020;20(4):23-34. doi:10.32947/ajps.v20i4.771.

Authors

Fatemeh Sadat Ahmadi
Majid Morovati-Sharifabad
mmorovati@ardakan.ac.ir (Primary Contact)
Gholam Hossein pourghanbari Marvast
Elham Salehi
Ehsan Zarei MahmoudAbadi
1.
Ahmadi FS, Morovati-Sharifabad M, pourghanbari Marvast GH, Salehi E, Zarei MahmoudAbadi E. Hydroalcoholic Extract of Cuscuta chinensis Suppresses Angiogenesis and Enhances Paclitaxel Activity in the Chick Chorioallantoic Membrane (CAM) Model: Cuscuta Anti Angiogenic. Arch Breast Cancer [Internet]. [cited 2026 May 31];13(3). Available from: https://archbreastcancer.com/index.php/abc/article/view/1296

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