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Flavonoid-Rich Extract of Oldenlandia diffusa (Willd.) Roxb. Inhibits Gastric Cancer by Activation of Caspase-Dependent Mitochondrial Apoptosis

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Chinese Journal of Integrative Medicine Aims and scope Submit manuscript

Abstract

Objective

To evaluate the apoptosis and cycle arrest effects of Oldenlandia diffusa flavonoids on human gastric cancer cells, determine the action mechanisms in association with the mitochondrial dependent signal transduction pathway that controls production of reactive oxygen species (ROS), and evaluate the pharmacodynamics of a mouse xenotransplantation model to provide a reference for the use of flavonoids in prevention and treatment of gastric cancer.

Methods

Flavonoids were extracted by an enzymatic-ultrasonic assisted method and purified with D-101 resin. Bioactive components were characterized by high-performance liquid chromatography. Cell lines MKN-45, AGS, and GES-1 were treated with different concentrations of flavonoids (64, 96, 128, 160 µg/mL). The effect of flavonoids on cell viability was evaluated by MTT method, and cell nuclear morphology was observed by Hoechst staining. The apoptosis rate and cell cycle phases were measured by flow cytometry, the production of ROS was detected by laser confocal microscope, the mitochondrial membrane potential (MMP) were observed by fluorescence microscope, and the expression of apoptotic proteins related to activation of mitochondrial pathway were measured by immunoblotting. MKN-45 cells were transplanted into BALB/c nude mice to establish a xenograft tumor model. Hematoxylin and eosin staining was used to reveal the subcutaneous tumor tissue. The tumor volume and tumor weight were measured, the expression levels of proliferation markers proliferating cell nuclear antigen (PCNA) and Ki-67 were detected by immunohistochemistry, and the expression levels of CA72-4 were measured by enzyme linked immunosorbent assay.

Results

Oldenlandia diffusa flavonoids inhibited proliferation of MKN-45 and AGS human gastric cancer cells, arrested the cell cycle in G1/S phase, induced accumulation of ROS in the process of apoptosis, and altered MMP. In addition, flavonoids increased Apaf-1, Cleaved-Caspase-3, and Bax, and decreased Cyclin A, Cdk2, Bcl-2, Pro-Caspase-9, and Mitochondrial Cytochrome C (P<0.05). The MKN-45 cell mouse xenotransplantation model further clarified the growth inhibitory effect of flavonoids towards tumors. The expression levels of PCNA and Ki-67 decreased in each flavonoid dose group, the expression level of CA72-4 decreased (P<0.05).

Conclusion

Flavonoids derived from Oldenlandia diffusa can inhibit proliferation and induce apoptosis of human gastric cancer cells by activating the mitochondrial controlled signal transduction pathway.

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Authors and Affiliations

  1. First School of Clinical Medicine, Gansu University of Chinese Medicine, Lanzhou, 730000, China

    Jia-yin Ling,  Qiu-lan Wang, Hao-nan Liang, Qing-bo Liu & Dong-hong Yin

  2. Gansu Provincial Hospital, Lanzhou, 730000, China

    Jia-yin Ling,  Qiu-lan Wang, Hao-nan Liang, Qing-bo Liu & Dong-hong Yin

  3. College of Pharmacy, Gansu University of Chinese Medicine, Lanzhou, 730000, China

    Li Lin

  4. Institute of Chinese (Tibetan) Medicine Resources, Gansu University of Chinese Medicine, Lanzhou, 730000, China

    Li Lin

Authors
  1. Jia-yin Ling
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  2. Qiu-lan Wang
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  3. Hao-nan Liang
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  4. Qing-bo Liu
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  5. Dong-hong Yin
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  6. Li Lin
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Contributions

Wang QL designed the study and guided the revision of the manuscript. Ling JY completed most of the experimental projects and wrote the manuscript. Yin DH assisted Ling JY in animal feeding and provided technical guidance for animal experiments. Lin L instructed Liang HN and Liu QB regarding flavonoid extraction. All authors read and approved the final version.

Corresponding author

Correspondence to Qiu-lan Wang.

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The authors declare no conflict of interest.

Additional information

Supported by the Key Research and Development Program of Gansu Province (No. 21YF5FA131), the Innovation Fund of Gansu Universities (No. 2021A-084), and the Graduate Innovation Fund of Gansu University of Chinese Medicine (No. 2021CX49)

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Flavonoid-rich Extract of Oldenlandia diffusa (Willd.) Roxb. Inhibits Gastric Cancer by Activation of Caspase-dependent Mitochondrial Apoptosis

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Ling, Jy., Wang, Ql., Liang, Hn. et al. Flavonoid-Rich Extract of Oldenlandia diffusa (Willd.) Roxb. Inhibits Gastric Cancer by Activation of Caspase-Dependent Mitochondrial Apoptosis. Chin. J. Integr. Med. 29, 213–223 (2023). https://doi.org/10.1007/s11655-022-3679-4

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  • DOI: https://doi.org/10.1007/s11655-022-3679-4

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