Ethanol extract of Ginger Zingiber officinale Roscoe by Soxhlet method induces apoptosis in human hepatocellular carcinoma cell line

Abstract

Introduction: Ginger has had a long history of use in healthcare in Asian countries, including Vietnam. Internationally, there have been a few studies done to assess the anticancer activity and potency of ginger. In this study, we investigated the activity (cytotoxicity) of various ginger root extracts that were produced by different preparation methods on human hepatocellular carcinoma cell line (HepG2 cell line). Methods: Ginger (Zingiber officinale Roscoe) root extract were extracted from the fresh ginger roots by three methods, which included maceration (G1), Soxhlet (G3), and ultrasonic (G5) methods. The extracts were then evaluated for cytotoxic activity on HepG2 hepatocellular carcinoma cell line based on IC 50 calculation. The most potent extract was then evaluated for its cytotoxicity by Propidium iodide (PI) staining method and microscopic examination by fluorescence microscopy for both 2D and 3D culture conditions. After treatment, cells were also stained with Annexin-V to assess the apoptosis-inducing ability of the extract. Results: The results showed that the ginger root extract prepared by the Soxhlet method (G3) had the strongest activity, and showed significant differences compared to extracts from methods G1 and G5, on the hepatocellular carcinoma cell lines cultured in both 2D and 3D conditions. Indeed, the IC 50 value of ginger root extract of the G3 was 83.3 ± 0.9189 µg/ml, compared to 159 ± 7.6 µg/ml of G1, and 284 ± 5.116 µg/ml of G5 in 2D culture latform; the IC 50 value was 228 ± 33.52µg/ml for G3 compared to 341 ± 3.93 µg/ml for G1, and 603.7 ± 56.33 µg/ml for G5 in 3D culture platform. The results of PI staining of cells in 2D and 3D culture conditions showed that ginger root extract killed HepG2 cells in a concentration-dependent manner. The nuclei of HepG2 cells in 2D culture showed nuclear fragmentation when treated with ginger root extract; this is one of the apoptosis indicators. Moreover, Annexin-V staining results showed that HepG2 cells underwent apoptosis when treated with ginger root extract. Conclusion: The results of the study show that ginger root extract prepared by the Soxhlet method induced strong cytotoxicity of hepatocellular carcinoma cells cultured in both 2D and 3D conditions, through induction of apoptosis. The evidence suggests that ginger is a potential agent for anticancer therapy. It is necessary to perform further research to isolate compounds from fresh ginger root extract by the Soxhlet method to evaluate the mechanism of anti-tumor activity.

Chemical constituents and biological activities of different extracts from ginger plant (Zingiber officinale)

“…These results may be due to the higher molecular weight compounds of released molecules during boiling of hot water extract and to the compounds identified by HPLC with anticancer activity. These results were in an agreement with Nguyen et al [64] and it's may be used in a form of tablets in the treatment of a wide range of other cancers, including lung, bladder, cervical, various GL cancers (gastric, pancreatic, liver, colorectal) and ovarian cancers which may be also in an agreement with Marrelli et al [43] as well as Prasad and Tyagi [65] due to its higher antiradical activity and it may be used in the form of medication to capture free radicals once it enters the body. It was shown that ginger exhibited an important role in cancer prevention by activating and inactivating various molecular pathways as reported by Rahmani et al [66].…”

Evaluation of antioxidant capacity, structure, and surface morphology of ginger (Zingiber officinale) using different extraction methods

“…The extract obtained with the US and AM methods presented significantly lower antioxidant activity (p < 0.05) than the M, R1, R2, and R3 methods, due to shorter exposure time (40 min) and heat stress [ 14 ]. Moreover, according to Nguyen et al [ 29 ], cavitation extraction is not strong enough to break the cell walls and release substances with antioxidant capacity.…”

Phytochemical Profiling and Antioxidant Activities of Red Ginger (Zingiber officinale var. rubrum) Cultivated Eco-Farming

“…Extracts were evaporated under vacuum to dryness and stored in dried bottles at 4°C. 18 Total Flavonoid Content One millilitre of the Simplicia sample was dissolved in 10 mL water and filtered. The filtrate is put into a test tube.…”