Effects of barakol from Cassia siamea on neuroblastoma SH-SY5Y cell line: A potential combined therapy with doxorubicin.

Management of neuroblastoma is challenging because of poor response to drugs, chemotherapy resistance, high relapse, and treatment failures. Doxorubicin is a potent anticancer drug commonly used for neuroblastoma treatment. However, doxorubicin induces considerable toxicities, particularly those caused by oxidative-related damage. To minimize drug-induced adverse effects, the combined use of anticancer drugs with natural-derived compounds possessing antioxidant properties has become an interesting treatment strategy. Barakol is a major compound found in Cassia siamea, an edible plant with antioxidant and anticancer properties. Therefore, barakol could potentially be used in combination with doxorubicin to synergize the anticancer effect, while minimizing the oxidative-related toxicities. Herein, the potential of barakol (0.0043-43.0 µM) to synergize the anticancer effect of low-dose doxorubicin (0.5 and 1.0 µM) was investigated. Results indicated that barakol could enhance the cytotoxic effect of low-dose doxorubicin by affecting the cell viability of the treated cells. Furthermore, the co-treatment with barakol and low-dose doxorubicin decreased the levels of intracellular ROS when compared with the control. Moreover, the antimetastatic effect of the barakol itself was studied through its ability to inhibit metalloproteinase-3 (MMP-3) activity and prevent cell migration. Results revealed that the barakol inhibited MMP-3 activity and prevented cell migration in time- and dose-dependent manners. Additionally, barakol was a non-cytotoxic agent against the normal tested cell line (MRC-5), which suggested its selectivity and safety. Taken together, barakol could be a promising compound to be further developed for combination treatment with low-dose doxorubicin to improve therapeutic effectiveness but decrease drug-induced toxicities. The inhibitory effects of barakol on MMP-3 activity and cancer cell migration also supported its potential to be developed as an antimetastatic agent.

13-Butoxyberberine Bromide Inhibits Migration and Invasion in Skin Cancer A431 Cells.

Cancer metastasis is the primary cause of cancer morbidity and mortality. Anti-metastasis mechanism of skin cancer by 13-butoxyberberine bromide, a novel berberine derivative, has not yet been reported. This study investigated the effects of 13-butoxyberberine bromide on migration and invasion of skin cancer A431 cells. The cytotoxicity of 13-butoxyberberine bromide was determined by MTT assay. The effect of 13-butoxyberberine bromide on cell migration and invasion were examined using a wound-healing assay, transwell migration assay, and transwell invasion assay, respectively. The cell adhesion ability was determined by an adhesion assay. Protein expressions that play important roles in cancer migration and invasion were evaluated by Western blot analysis. The results showed that 13-butoxyberberine bromide effectively inhibited cell migration, invasion, and adhesion in A431 cells. Interestingly, 13-butoxyberberine bromide was more effective for cell migration inhibition than berberine. In addition, 13-butoxyberberine bromide showed anti-migration and anti-invasion effects by down-regulated MMP-2 and MMP-9 expression and up-regulated TIMP-1 and TIMP-2 expression in A431 cells. Moreover, pretreatment with 13-butoxyberberine bromide significantly inhibited EGF-induced cell migration and p-EGFR, ERK, p-ERK, STAT3, and p-STAT3 expressions in A431 cells at lower concentrations when compared with the berberine. These findings indicated that 13-butoxyberberine bromide could be further developed as an anticancer agent.

Mangosteen for malignancy prevention and intervention: Current evidence, molecular mechanisms, and future perspectives.

Mangosteen (Garcinia mangostana L.), also known as the "queen of fruits", is a tropical fruit of the Clusiacea family. While native to Southeast Asian countries, such as Thailand, Indonesia, Malaysia, Myanmar, Sri Lanka, India, and the Philippines, the fruit has gained popularity in the United States due to its health-promoting attributes. In traditional medicine, mangosteen has been used to treat a variety of illnesses, ranging from dysentery to wound healing. Mangosteen has been shown to exhibit numerous biological and pharmacological activities, such as antioxidant, anti-inflammatory, antibacterial, antifungal, antimalarial, antidiabetic, and anticancer properties. Disease-preventative and therapeutic properties of mangosteen have been ascribed to secondary metabolites called xanthones, present in several parts of the tree, including the pericarp, fruit rind, peel, stem bark, root bark, and leaf. Of the 68 mangosteen xanthones identified so far, the most widely-studied are α-mangostin and γ-mangostin. Emerging studies have found that mangosteen constituents and phytochemicals exert encouraging antineoplastic effects against a myriad of human malignancies. While there are a growing number of individual research papers on the anticancer properties of mangosteen, a complete and critical evaluation of published experimental findings has not been accomplished. Accordingly, the objective of this work is to present an in-depth analysis of the cancer preventive and anticancer potential of mangosteen constituents, with a special emphasis on the associated cellular and molecular mechanisms. Moreover, the bioavailability, pharmacokinetics, and safety of mangosteen-derived agents together with current challenges and future research avenues are also discussed.

Zanthoisobutylamides A - C: rare dimeric C-6 substituent dihydrobenzophenanthridine alkaloids from the roots of Zanthoxylum nitidum.

Three new dihydrobenzophenantridine alkaloids, zanthoisobutylamides A-C (2-4), consisting of a rare 6-alkylamide dihydrochelerythrine moiety, and two new small molecules of the unsaturated alkylamide, zanthoxylumamide J (1) and of phenylpropanoid, methyl 2-hydroxy-3,4-dimethoxycinnamate (5) together with 44 known compounds were isolated from the roots of Zanthoxylum nitidum. The structures of these compounds were established by analysis of spectroscopic data and comparison of their spectroscopic data with those previously published data. Some isolated compounds were evaluated for their cytotoxic activities.

A new alkaloid and a new benzaldehyde derivative from the twig of Zanthoxylum rhetsa (Roxb.) DC.

A new alkaloid, 2-acetyl-4-methoxyfuro[2,3-b]quinoline (1), and a new benzaldehyde derivative, (2'S)-4-(2'-hydroxy-3'-methyl-3'-butenoxy)benzaldehyde (2), were isolated from the twig of Zanthoxylum rhetsa (Roxb.) DC. along with twenty-six known compounds (3-28). Their structures were determined by spectroscopic analysis (1D and 2D NMR spectroscopy and HRMS analysis) and comparison with data reported in the literature. Thirteen of the known compounds were evaluated for their cytotoxic activities against human cancer cell lines that included MDA-MB-231, SW1353, A549, and HCT116. (±)-8-Acetonyldihydronitidine (15) showed moderate cytotoxicity toward the SW1353 cancer cell line with an IC50 value of 18.90±0.39 µg/mL, and exhibited weak cytotoxic activity against MDA-MB-231, A549 and HCT116 cell lines with IC50 values of 49.86-71.32 µg/mL.

New acylphloroglucinols from a crude acetone extract of Eucalyptus camaldulensis Dehnh. leaf.

Acylphloroglucinols are well-known Eucalyptus secondary metabolites which exhibit a variety of structures and bioactivities. The investigation of a crude acetone extract of Eucalyptus camaldulensis leaves led to the isolation of two new acylphloroglucinols, eucalypcamals O and P (1 and 2) together with seven phloroglucinols (3-9), and a benzene derivative (10). Their chemical structures were elucidated by 1D and 2D nuclear magnetic resonance (NMR) spectroscopy and mass spectroscopy. The absolute configurations of compounds 1 and 2 were established by comparison of experimental and calculated electronic circular dichroism (ECD) data. In the putative biosynthetic pathway, eucalypcamals O and P should be derived from hetero-Diels-Alder reaction between grandinol and trans-isoeugenol.

Effects of Artonin E on Cell Growth Inhibition and Apoptosis Induction in Colon Cancer LoVo and HCT116 Cells.

Today, colon cancer is the leading cause of cancer death. In Thailand, colon cancer is the third most common cancer in men and the second in women. Currently, the treatments for colon cancer include surgery, chemotherapy, radiation therapy, immunotherapy, hormone therapy, targeted drug therapy, and stem cell therapy. However, some treatments have side effects for cancer patients, causing unwanted symptoms. In addition, targeted therapy comes with a high cost for patients. Therefore, bioactive compounds might be a good choice for colon cancer treatment. In this study, we investigated the effect of artonin E on apoptosis induction in colon cancer LoVo and HCT116 cells. The concentration ranges of artonin E at 3, 5, 10, and 30 µg/mL in LoVo cells and 1, 1.5, 2, and 3 µg/mL in HCT116 cells were examined. The results implied that artonin E decreased cell viability and increased apoptotic cells in a dose-dependent manner. In addition, artonin E stimulated mitochondrial membrane potential (ΔΨm) changes associated with apoptosis by increasing the sub-G1 population analyzed by flow cytometry. Western blotting showed that artonin E increased the proapoptotic protein, Bax, and decreased anti-apoptotic proteins' (Bcl-2 and Bcl-x) expression. Moreover, artonin E also increased cleaved caspase-7 and cleaved-PARP expression in both LoVo and HCT116 cells. Interestingly, artonin E induced apoptosis through p-ERK1/2, p-p38/p38, and p-c-Jun expression in both cells. Our results suggested that artonin E induced apoptosis via caspase activation associated with the MAPKs signaling pathway. Therefore, artonin E might be used as a potential anticancer drug for colon cancer in the future.

Phloroglucinol-meroterpenoids from the leaves of Eucalyptus camaldulensis Dehnh.

Fourteen undescribed phloroglucinol-meroterpenoids, namely eucalypcamals A-N, were isolated from a CH2Cl2 extract of the leaves of Eucalyptus camaldulensis Dehnh. In addition, from the same extract, twelve known phloroglucinols, three known flavonoids, and four known phenolic compounds were also isolated. The structures of the undescribed compounds were analyzed by 1D and 2D nuclear magnetic resonance (NMR) spectroscopy, and high resolution electrospray ionization mass spectrometry (HRESIMS). The assignments of the absolute configurations were performed by comparing the experimental electronic circular dichroism (ECD) data with the calculated values. Eucalyprobusal E was found to be cytotoxic against HCT116, Jurkat, and MDA-MB-231 cell lines with IC50 values of 17.6, 9.44, and 17.9 µM, respectively. Eucalrobusone F exhibited antibacterial activity against methicillin-resistant S. aureus (MRSA) and S. aureus with minimum inhibitory concentration/minimum bactericidal concentration (MIC/MBC) values of 4/4 µg/mL while euglobal Ia1 showed antifungal activity with MIC/MFC values of 16/16 µg/mL.

Enhancing Anticancer Potency of a 13-Substituted Berberine Derivative with Cationic Liposomes.

Cationic liposomal formulations of the telomeric G-quadruplex stabilizing ligand, 13-(2-naphthylmethoxy)berberine bromide (1), have been developed with the purpose of delivering 1 into the nucleus of cancer cells for potential telomere targeting. Berberine derivative 1 was encapsulated in various cationic lipids 2-4 by the thin film evaporation method; these lipids are cationic after amine protonation. The most appropriate liposomal berberine formulation was that of 1 and the cholesterol derived cationic lipid 4 in a weight ratio of 1 : 20 with 76.5% encapsulation efficiency of 1. Cellular uptake studies in the HeLa and HT-29 cancer cells lines showed that the liposomal berberine derivative uptake in the cells was higher and more stable than for berberine derivative 1 alone while free 1 was completely decomposed in the cells within 60 min exposure to the cells. Anticancer activity of the liposomal berberine derivative 1 based on 4 was greater than that for the free berberine derivative 1 in the MCF-7, HeLa and HT-29 cell line by 2.3-, 4.9- and 5.3-fold, respectively, and also, interestingly, superior to the anticancer drug doxorubicin against the HT29 cancer cell line.

Apoptosis Induction through MAPK Signaling Pathway in LoVo Cells by Fatty Acid Fraction from Rice Bran Oil.

Colorectal cancer is one of the five leading cancer incidents and mortality in Thailand and worldwide. Fatty acids (FA) are bioactive molecules which have potential as adjunctive chemotherapeutic agents. To study the effect of fatty acid fraction (FAs) extracted from organic rice bran oil on apoptosis induction and growth inhibition in human colorectal cancer cell line, LoVo cells. The results demonstrated that FAs inhibited cell viability and induced cell death via apoptosis associated with MAPKs pathway. The EC50 of FAs in LoVo was 172.80 ± 1.05 µg/ml. FAs treatment significantly increased nuclear condensation and decreased mitochondrial membrane potential. Moreover, FAs activated Bax, Caspase-9, -7 and PARP cleavage, while inhibited Bcl-2 expression. Furthermore, FAs increased p53 expression and phosphorylation of ERK and p38. FAs extracted from organic rice bran oil inhibited LoVo cell viability and induced apoptosis via MAPKs pathway. These data suggest the potential use of FAs extracted from organic rice bran oil to prevent or treat colon cancer in the future.

A new lignan from the stem bark of Fagraea fragrans Roxb.

A new lignan, named fagraeanolide (1), and 14 known compounds were isolated from the stem bark of Fagraea fragrans Roxb. Their structures were determined by spectroscopic methods. Fagraeanolide is the first identified oxofurofuran lignan from the genus Fagraea, whileß-boswelic acid (4), gentiogenol (5), 3-(4-hydroxy-3-methoxyphenyl)-acrylic acid octacosyl ester (7) and pinoresinol (14) were isolated from this plant for the first time. The crude extract of F. fragrans was not toxic to cell lines. The isolated compounds showed no antibacterial activity.

Protective effect of purple corn silk extract against ultraviolet-B-induced cell damage in human keratinocyte cells.

Ultraviolet-B (UVB) could lead to inflammation and cell death induction. Purple corn silk (PCS), part of female flower of corn has multiple pharmacological properties. This investigation focused on determining the preventive effects of PCS extract on human keratinocyte HaCaT cell damage induced by UVB irradiation. Cells were irradiated with 25 mJ/cm2 UVB after pre-treated with PCS extract for 1 h. Then, the cells were then placed in culture medium followed by subsequent experiments. Cell survival was determined by MTT assay. The immunofluorescence, DCFH-DA, JC-1, and Hoeshst33342 staining assays were used to determine γ-H2AX, intracellular reactive oxygen species (ROS), membrane potential of mitochondria, and nuclear condensation, respectively. Western blot analysis was used to investigate the proteins expression. The statistically significant comparison was calculated by analysis of variance at P < 0.05. The fluorescence and protein band intensity were quantified by Image J densitometer. The results indicated cell survival was increased upon PCS extract pretreatment followed by UVB exposure. PCS extract decreased γ-H2AX expression, intracellular ROS overproduction, and nuclear condensation in cells induced by UVB. Furthermore, The PCS extract pretreatment attenuated apoptosis response through stabilized mitochondrial membrane potential, decreased apoptosis mediator proteins including Bax, Bak, cleaved-caspases, and cleaved-PARP, and increased Bcl-2 and Bcl-xL expression comparing to the UVB-treated control. This finding demonstrated that the PCS extract can reduce the deleterious effects from UVB exposure through decreased intracellular ROS, DNA damage, and apoptosis induction on HaCaT cells.

Corrigendum to "Interaction between goniothalamin and peroxisomal multifunctional enzyme type 2 triggering endoplasmic reticulum stress" [Heliyon 6 (10) (October 2020) e05200].

[This corrects the article DOI: 10.1016/j.heliyon.2020.e05200.].

Inhibitory effects on chondrosarcoma cell metastasis by Senna alata extract.

BACKGROUND: Senna alata L. Roxb or candle bush is a traditional medicinal plant with a wide range of biological activities including anti-inflammatory, antimicrobial and antifungal. Leaf extract of S. alata showed the anti-tumor activity in various cancer cell lines. In this study, we focused on the inhibitory mechanism of S. alata extract (SAE) on cancer metastasis including cell migration, cell invasion and signaling pathways in chondrosarcoma SW1353 cells. PURPOSE: This study aimed to evaluate the anti-metastatic mechanisms of Senna alata extract on chondrosarcoma SW1353 cells. METHODS: Screening for phytochemicals in biologically active fraction of SAE was analysed by 1H NMR spectroscopy. Cell viability and cytoxicity were determined by using MTT assay. Cell migration was observed by scratch wound healing and transwell migration assay. Cell invasion and cell adhesion assay were examined by Matrigel coated transwell chambers or plates. The expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), MAPKs and PI3K/Akt signaling pathways and NF-κB were detected by Western blot analysis. RESULTS: The SAE treatment at the sub-cytoxic and non-cytotoxic concentrations significantly inhibited cell migration, cell invasion and cell adhesion of SW1353 cells in a dose-dependent manner. The results from Western blot analysis showed decreased MMP-2 and MMP-9 expression, while increased TIMP-1 and TIMP-2 expression in SAE treated cells. Moreover, SAE suppressed phosphorylation of ERK1/2, p38 and Akt but decreased NF-κB transcription factor expression in SW1353 cells. CONCLUSION: These results revealed that SAE could reduce MMP-2 and MMP-9 expression by downregulation of NF-κB which is downstream of MAPKs and PI3K/Akt signaling pathway in SW1353 cells resulting in reduced cancer cell migration and invasion. Therefore, SAE may have the potential use as an alternative treatment of chondrosarcoma metastasis.

New flavonoids and xanthone from the stem bark of Artocarpus rigidus blume and cytotoxicity.

Three new flavonoids named artorigidinones A-C and a new xanthone named artorixanthone together with seven known compounds were isolated from the stem bark of Artocarpus rigidus Blume. Their structures were characterized by spectroscopic data. γ-Geranylapigenin exhibited cytotoxicity to a fibroblast-like cell line (SW1353) (IC50 < 0.32 µg/mL) stronger than a standard drug.

Interaction between goniothalamin and peroxisomal multifunctional enzyme type 2 triggering endoplasmic reticulum stress.

Endoplasmic reticulum stress is one of the pathways involved in cell cytotoxicity. In this study, goniothalamin, one of styryllactone compounds found in plant Goniothalamus spp., was observed to trigger ER stress in HeLa cell line. In addition, we demonstrated that peroxisomal multifunctional enzyme type2 (MFE2) was a specific goniothalamin-binding protein using an in vitro goniothalamin-linked bead pull-down assay. Since MFE2 has been reported to be an important mediator enzyme for peroxisomal ß-oxidation of a very long chain fatty acid metabolism, therefore computational molecular docking analysis was performed to confirm the binding of goniothalamin and MFE2. The results indicated that goniothalamin structure binds to scp-2 domain, enoyl-CoA hydratase 2 domain and (3R)-hydroxyacyl-CoA dehydrogenase domain of MFE2. To further determine the effect of MFE2 on ER stress induction, MFE2 knockdown by siRNA in HeLa cell was conducted. The results implied that MFE2 triggered CHOP, a key mediator of ER stress-induced apoptosis, expression. Therefore, these data inferred that goniothalamin may interrupt the MFE2 function resulting in lipid metabolism perturbation associated with ER stress-independent activation of unfolded protein response. This is the first report to show that goniothalamin binds directly to MFE2 triggering ER stress activation probably through the lipid metabolism perturbation.

Antimetastatic Potential of Rhodomyrtone on Human Chondrosarcoma SW1353 Cells.

Chondrosarcoma is primary bone cancer, with the forceful capacity to cause local invasion and distant metastasis, and has a poor prognosis. Cancer metastasis is a complication of most cancers; it is one of the leading causes of cancer-related death. Rhodomyrtone is a pure compound that has been shown to induce apoptosis and antimetastasis in skin cancer. However, the inhibitory effect of rhodomyrtone on human chondrosarcoma cell metastasis is largely unknown. Effect of rhodomyrtone on cell viability in SW1353 cell was determined by MTT assay. Antimigration, anti-invasion, and antiadhesion were carried out to investigate the antimetastatic potential of rhodomyrtone on SW1353 cells. Gelatin zymography was performed to determine matrix metalloproteinase-2 (MMP-2) and MMP-9 activities. The effect of rhodomyrtone on the underlying mechanisms was performed by Western blot analysis. The results demonstrated that rhodomyrtone reduced cell viability of SW1353 cells at the low concentration (<3 µg/mL); cell viability was >80%. Rhodomyrtone at the subcytotoxic concentrations (0.5, 1.5, and 3 µg/mL) significantly inhibited cell migration, invasion, and adhesion of SW1353 cells in a dose-dependent fashion. Protein expression of integrin αv, integrin ß3, and the downstream migratory proteins including focal adhesion kinase (FAK) and the phosphorylation of serine/threonine AKT, Ras, RhoA, Rac1, and Cdc42 were inhibited after treatment with rhodomyrtone. Moreover, we found that rhodomyrtone decreased the protein level of MMP-2 and MMP-9 as well as the enzyme activity in SW1353 cells. Meanwhile, tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 expression was increased in a dose-dependent fashion. Besides, rhodomyrtone dramatically inhibited the expression of growth factor receptor-bound protein-2 (GRB2) and the phosphorylated form of extracellular signal regulation kinase1/2 (ERK1/2) and c-Jun N-terminal kinase1/2 (JNK1/2). These results indicated that rhodomyrtone inhibited SW1353 cell migration, invasion, and metastasis by suppressing integrin αvß3/FAK/AKT/small Rho GTPases pathway as well as downregulation of MMP-2/9 via ERK and JNK signal inhibition. These findings indicate that rhodomyrtone possessed the antimetastasis activity that may be used for antimetastasis therapy in the future.

Artonin E sensitizes TRAIL-induced apoptosis by DR5 upregulation and cFLIP downregulation in TRAIL-refractory colorectal cancer LoVo cells.

BACKGROUND: The TRAIL treatment is an ideal strategy for colorectal cancer (CRC) therapy because of minimal collateral damage to normal cells. Unfortunately, some CRC is TRAIL-refractory cancer, such as LoVo cells. In an effort to overcome TRAIL-refractory cancer, we investigated the effect of artonin E in regulating death receptor 5 (DR5) and cellular FLICE (FADD-like IL-1ß-converting enzyme)-inhibitory protein (cFLIP), two major mediators regulate TRAIL-induced apoptosis, in LoVo cells as a model of TRAIL refractory CRC. METHODS: TRAIL-refractory cancer (LoVo cells) was treated with artonin E and TRAIL. Cell viability was determined by MTT assay. Apoptotic chromatin condensation was observed by fluorescent Hoechst33342 staining. The mRNA and protein expression of DR5 and FLIP was determined by quantitative PCR and Western blotting analysis, respectively. RESULTS: The combination treatment of artonin E and TRAIL enhanced cytotoxicity and apoptotic chromatin condensation in LoVo cells significantly, while treatment of artonin E or TRAIL alone was not. Artonin E enhanced both mRNA and protein expression of DR5. Interestingly, this is the first report showing that artonin E decreased protein expression of cFLIP. All together we showed that artonin E enhanced TRAIL-induced apoptosis in LoVo cells through DR5 upregulation and cFLIP downregulation. CONCLUSIONS: Artonin E was able to increase DR5 expression and decrease cFLIP expression in LoVo cells. These results showed that LoVo cells sensitized TRAIL-induced apoptosis in combined treatment with artonin E and TRAIL. Therefore, the combination treatment of artonin E and TRAIL is one of the potential strategies used for TRAIL-refractory CRC therapy in the future.

Cytotoxic xanthones from the roots of Mesua ferrea L.

Five undescribed xanthones, 4-methoxypyranojacareubin, 4-hydroxy-3-prenylpyranoxanthone, 1-hydroxy-5,7-dimethoxyxanthone, 5-hydroxy-1,6,7-trimethoxyxanthone and 2-hydroxy-1,5-dimethoxyxanthone, together with thirty-three known xanthones were isolated from the roots of Mesua ferrea L. The structures of all isolated xanthones were elucidated based on spectroscopic methods. 5-Hydroxy-1,6,7-trimethoxyxanthone and 2-hydroxy-1,5-dimethoxyxanthone were also confirmed by X-ray diffraction data. In addition, the isolated compounds were determined for antibacterial, antioxidant and cytotoxic activities. The known 1,5,6-trihydroxyxanthone showed cytotoxicity against A375, PC-3 and HaCaT cell lines with IC50 values of 5.73 µg/mL, 5.93 µg/mL and 8.94 µg/mL, respectively.