β-Dihydroagarofuran-type sesquiterpenoids from the stems of Celastrus orbiculatus with their cytotoxic activities

A new β-dihydroagarofuran-type sesquiterpenoid named 1α,2α,5α,11-tetraacetoxy-8α-(trans-p-coumaroyl)-β-dihydroagarofuran (1), together with five known compounds (2-6) were isolated from the CHCl3-soluble extract of the stems of Celastrus orbiculatus. The structure of new compound was elucidated with spectroscopic physico-chemical analyses. All isolates were evaluated for in vitro cytotoxic activity against four human cancer lines including HepG2, MCF-7, A549 and HCT-116 cells. Among them, compounds 1 and 6 showed potent cytotoxic activities on HepG2 cells with IC50 values of 8.78 ± 2.31 and 10.28 ± 1.15 μM, respectively. In addition, compound 6 exhibited significant cytotoxic activity on HCT-116 cells with IC50 value of 6.37 ± 2.52 μM

Studies on the effect of Celastrus orbiculatus (Celastraceae) extract on chemosensitivity of liver cancer cells via Wnt/β-catenin pathway

Purpose: To examine the efficacy of Celastrus orbiculatus extract (COE) on the chemosensitivity of liver cancer (LC) cells and its mechanism of action.Methods: Hep G2/ADM cells in the logarithmic growth phase were assigned to a control group (no treatment for cell culture medium only) and a study group (120 μg/ml COE added to the culture medium). After 48 h of incubation, the biological responses were compared. The study group wasdivided into groups A and B, while control group was divided into groups C and D, with 1 μmol/L XAV939 added in groups A and C. Cell proliferation, cell invasion, cell apoptosis rate, and apoptosis protein in the four groups were evaluated.Results: The study group showed significantly lower values in terms of cell proliferation and cell invasiveness (p < 0.05) and a higher apoptotic rate than the control group (p < 0.05)). The study group also demonstrated an elevated pro-apoptotic protein Bax level and a declined anti-apoptotic protein Bcl-2 level. In contrast to group B, the proliferation and invasiveness of Hep G2/ADM cells in group A treated with the inhibitor, XAV939, were significantly lower (p < 0.05), while the apoptotic rate exhibited a significant increase (p < 0.05). There was a rise in the level of pro-apoptotic protein, Bax, and a fall in the anti-apoptotic protein Bcl-2 level in group A. Lower levels of β-catenin, c-Myc, and cyclin D1 protein were observed in the study group compared with the control group (p < 0.05). Compared with other groups, the multiplication capacity and invasiveness of cells in group A treated with COE and inhibitor XAV939 significantly declined, while the apoptotic rate increased (p < 0.05).Conclusion: COE reverses drug resistance in chemotherapy by inhibiting the expression of Wnt/β-catenin pathway in LC cells. Therefore, COE has potentials for use along with chemotherapeutic agents in the management of liver cancer.

Improvement of the Chemosensitivity of Gastric Carcinoma Cells by Celastrus orbiculatus Extracts

Gastric carcinoma is one of the most prevalent malignancies with high morbidity and mortality. While chemotherapy is the major means for the management gastric carcinoma, tumors gradually exhibit drug resistance. Therefore, there is a need for agents capable of enhancing the sensitivity of tumor cells to chemotherapy. Herein, we have examined the effect of Celastrus orbiculatus extracts on chemosensitivity of drug resistant gastric carcinoma cells. Human gastric carcinoma cells (MGC-803 and SGC-7901) were cultured in the presence of cisplatin for the selection of drug resistant gastric cells. Next, the effect of C. orbiculatus extracts on multiplication capacity of drug resistant MGC-803 and SGC-7901 cells was examined by a variety of measures. Following C. orbiculatus extract treatment, the multiplication and invasiveness of drug resistant MGC-803 and SGC-7901 cells declined remarkably, with increased apoptosis and decreased levels of β-catenin, c-Myc, and cyclin D1 proteins, protein markers critical to cell proliferation, differentiation, and apoptosis. In summary, C. orbiculatus extract can effectively improve the sensitivity of cisplatin resistant gastric carcinoma cells to cisplatin and promote the apoptosis of tumor cells through the inhibition of the expression of proteins associated with the Wnt/β-catenin axis.

The Ethyl Acetate Extract From Celastrus orbiculatus Promotes Apoptosis of Gastric Cancer Cells Through Mitochondria Regulation by PHB

Objective: To investigate the effect of ethyl acetate extract from Celastrus orbiculatus (COE) on gastric cancer cell apoptosis and reveal its underlying molecular mechanism. In addition, it was aimed to stablish a theoretical basis for the clinical application of Celastrus orbiculatus in the gastric cancer treatment.Material and Methods: Western blot and RT-qPCR were used to detect mRNA and protein expression of PHB in gastric cancer and adjacent tissues. MTT method was used to detect the COE effect on the proliferation of AGS cells and to determine the 50% inhibitory concentration COE on these cells. COE effect on AGS apoptosis was evaluated by flow cytometry. Changes in apoptosis-related proteins expression in AGS cells were detected by western blot and changes in mitochondrial membrane potential were detected by JC-1 fluorescence staining. PHB expression was knocked down in AGS cells by lentiviral-mediated RNA interference. The COE antitumor effect was assessed in vivo using a subcutaneous transplantation tumor model in nude mice and in vivo fluorescence tracing technique in small animals.Results: The clinical samples analysis results showed that the PHB expression in gastric cancer samples was significantly higher than in corresponding adjacent tissues. MTT results showed that the AGS cell proliferation was significantly inhibited. RT-qPCR and western blot results showed that COE can significantly inhibit the PHB mRNA and protein expression, respectively. Flow cytometry analysis showed that COE was able to significantly promote AGS cell apoptosis. Western blot results also indicated that apoptosis-related protein expression changed significantly; BCL-2 expression significantly reduced while the Caspase-3 and Bax expression significantly increased after COE treatment. JC-1 fluorescence staining results showed that COE changed the mitochondrial membrane potential and activated the mitochondrial apoptosis pathway. Furthermore, in vivo experiments results demonstrated that the growth of subcutaneous transplanted tumor was significantly inhibited by the PHB knockdown and by the COE intragastric administration.Conclusion: COE can significantly promote apoptosis of human gastric cancer cells, which can be achieved by inhibiting PHB expression, thus altering the structure and function of mitochondria and activating the mitochondria apoptosis pathway. The antitumor effect of COE has also been proved in vivo.

Celastrus orbiculatus Celastraceae Thunb extracts inhibit proliferation and migration of oral squamous cell carcinoma cells by blocking NF-κB pathway

Purpose: To investigate the mechanism of action of Celastrus orbiculatus extract (COE) on oral squamous carcinoma cells. Methods: Tca8113 cells were divided into negative control and three COE treatment groups, viz, 20, 40, and 80 μg/mL of COE. Succinate dehydrogenase activity assay (MTT assay) and flow cytometry were used to assess cell proliferation and apoptosis. Cell migration and invasion were assessed by Transwell chamber and wound healing assays while relative protein expression was determined by Western blot. Results: As the concentration of COE increased, the number of cells arrested in the G0/G1 phase increased (p < 0.05), expressions of cell-cycle-related proteins decreased (p < 0.001); the number of apoptotic cells increased (p < 0.001), and the rates of cell migration and invasion decreased (p < 0.001). Exogenous COE significantly inhibited p-IκBα accumulation in the cytoplasm and induced IκBα accumulation. Nuclear p65 recruitment was reduced in cells treated with COE compared to untreated control cells (p < 0.001), suggesting that the classical NF-κB pathway was blocked by COE. Conclusion: These results demonstrate that COE inhibits the proliferation and migration of oral squamous cell carcinoma cells while promoting apoptosis by blocking NF-κB pathway. These findings suggest that Celastrus orbiculatus extract possesses a potential therapeutic effect on oral squamous cell carcinoma.

The biology of invasive alien plants in Canada. 14. Celastrus orbiculatus Thunb.

Celastrus orbiculatus Thunb. is an invasive alien liana native to northern regions of Japan, Korea, China and Russia. Since its intentional North American introduction in the 1870s, it is now found across eastern Canada and United States. Through an extensive adventitious root system and twining growth habit, C. orbiculatus smothers and girdles surrounding vegetation, drastically altering the environment and ecosystem processes. C. orbiculatus continues to be distributed as an ornamental. In addition, birds and small mammals consume its prolific fruit and facilitate novel introductions. C. orbiculatus is susceptible to glyphosate and triclopyr. Once established, however, management intensity is compounded by its extensive root system and continual resprouting and root-suckering. The climatic requirements across eastern Canada, ease of dispersal and rapid growth suggest that C. orbiculatus will continue to spread across its introduced range.

Celastrus orbiculatus Extract Reduces Stemness of Gastric Cancer Stem Cells by Targeting PDCD4 and EIF3H

Background Celastrus orbiculatus ethyl acetate extract (COE) has shown a strong anti-gastric cancer effect, but the understanding of its mechanism is still lacking. The results of previous studies indicated that COE may be able to inhibit the stemness of gastric cancer stem cells (GCSCs) by regulating PDCD4 and EIF3H expression. Aims To explore if COE could inhibit the stemness of GCSCs by regulating PDCD4 and EIF3H expression in vitro and in vivo. Procedure The GCSCs model was established by stem cell-conditioned culture. Spheroid formation and flow cytometry assays were used to detect the effect of COE on the spheroid formation ability of GCSCs and the percentage of CD44+/CD24+ and ALDH+ cell subpopulations. Western blot analysis was applied to measure the expression of GCSCs biomarkers (Nanog, Oct-4, and SOX-2), PDCD4, and EIF3H in GCSCs treated with COE; and RT-PCR was performed to investigate the effect of COE on PDCD4 mRNA expression in GCSCs. An in vivo tumorigenicity experiment was also conducted to evaluate the effect of COE on tumor-initiating ability of GCSCs in vivo; and the expression of PDCD4 and EIF3H in xenograft tissues was examined by immunohistochemistry (IHC) staining. Results After culture in stem cell-conditioned medium, SGC7901 cells manifested significantly enhanced spheroid formation ability, upregulated Nanog, Oct-4, and SOX-2 expression and increased percentages of CD44+/CD24+ and ALDH+ cell subpopulations, indicating successful establishment of the GCSCs model. COE treatment significantly inhibited the spheroid formation ability of GCSCs and reduced the percentage of CD44+/CD24+ and ALDH+ cell subpopulations. The western blot analysis showed a significant decrease of Nanog, Oct-4, SOX-2, and EIF3H expression and an increase of PDCD4 expression in GCSCs after COE treatment in a concentration-dependent manner. COE treatment also significantly upregulated the mRNA expression of PDCD4 in GCSCs. In addition, COE displayed a strong inhibitory effect on the tumor-initiating ability of GCSCs in vivo and upregulated PDCD4 and downregulated EIF3H expression in xenograft tissues. Conclusion COE may be able to inhibit GC growth by suppressing the stemness of GCSCs via regulating PDCD4 and EIF3H expression.