1.Cytotoxic and Apoptotic properties of the Flavonoid-rich Ethyl Acetate fraction of the Crude Methanol Leaf Extract of Syzygium samarangense (Blume) Merr. (Myrtaceae) against MCF-7 Breast Cancer Cells
Gerard Q. De Guzman ; Maria Stephanie Fay S. Cagayan ; Daisy-Mae A. Bagaoisan ; Shiela DV Miranda
Acta Medica Philippina 2020;54(Online):1-10
Introduction:
Breast cancer is the most common cancer among women in the Philippines and about 3 in every
100 Filipina will be diagnosed with breast cancer in their lifetime. There is a need to discover safe, yet inexpensive herbal extracts with potential cytotoxic properties as potential treatment modalities to treat breast cancer.
Objectives:
This study seeks to explore the cytotoxic and apoptotic properties of the ethyl acetate fraction of the defatted crude methanol leaf extract of Syzygium samarangense in MCF-7 breast cancer cell lines.
Methods:
Screening for flavonoids of the extracts was performed using TLC, total flavonoids, total phenols, FTIR and LC-MS spectroscopy. The hydrogen peroxide and ferric reducing anti-oxidant power were used as substrates to assess in vitro anti-oxidative properties of the extracts. The MTT dye viability assay was used to assess the cytotoxic properties of the extracts against MCF-7 cells. Apoptotic properties of the extracts in MCF-7 cells were determined by caspase-3 activation assay, DNA fragmentation patterns and fluorescence microscopy after annexin-V and propidium iodide staining.
Results:
The abundance of flavonoids in the ethyl acetate fraction of the crude methanol leaf extract was established by TLC, FTIR, LC-MS/MS, total flavonoid and total phenol analyses. The in vitro anti-oxidative properties of this extract was comparable to ascorbic acid. The median inhibitory concentration (IC50) of this extract in MCF-7 breast cancer cell lines was 7.2 mcg/mL while doxorubicin registered an IC50 of 1.2 mcg/mL. At this concentration, the extract was not cytotoxic to normally-dividing breast epithelial cells. Cytotoxicity of the extract was mediated via apoptosis as demonstrated by DNA fragmentation, caspase-3 activation and fluorescence microscopic analyses.
Conclusion
The study shows that the flavonoid-rich ethyl acetate fraction of the crude methanol leaf extract of
S. samarangense possesses potent apoptotic and cytotoxic properties against MCF-7 breast cancer cell lines at low concentrations.
MCF-7 Cells
;
Syzygium
2.Cytotoxic and Apoptotic Properties of the Flavonoid-rich Ethyl Acetate Fraction of the Crude Methanol Leaf Extract of Syzygium samarangense (Blume) Merr. (Myrtaceae) against MCF-7 Breast Cancer Cells
Gerard Q. De Guzman ; Maria Stephanie Fay S. Cagayan ; Daisy-Mae A. Bagaoisan ; Shiela DV Miranda
Acta Medica Philippina 2023;57(11):41-50
Introduction:
Breast cancer is the most common cancer among women in the Philippines and about 3 in every
100 Filipina will be diagnosed with breast cancer in their lifetime. There is a need to discover safe, yet inexpensive herbal extracts with potential cytotoxic properties as potential treatment modalities to treat breast cancer.
Objectives:
This study seeks to explore the cytotoxic and apoptotic properties of the ethyl acetate fraction of the
defatted crude methanol leaf extract of Syzygium samarangense in MCF-7 breast cancer cell lines.
Methods:
Screening for flavonoids of the extracts was performed using TLC, total flavonoids, total phenols, FTIR
and LC-MS spectroscopy. The hydrogen peroxide and ferric reducing anti-oxidant power were used as substrates to assess in vitro anti-oxidative properties of the extracts. The MTT dye viability assay was used to assess the cytotoxic properties of the extracts against MCF-7 cells. Apoptotic properties of the extracts in MCF-7 cells were determined by caspase-3 activation assay, DNA fragmentation patterns and fluorescence microscopy after annexin-V and propidium iodide staining.
Results:
The abundance of flavonoids in the ethyl acetate fraction of the crude methanol leaf extract was established by TLC, FTIR, LC-MS/MS, total flavonoid and total phenol analyses. The in vitro anti-oxidative properties of this extract was comparable to ascorbic acid. The median inhibitory concentration (IC50) of this extract in MCF-7 breast cancer cell lines was 7.2 mcg/mL while doxorubicin registered an IC50 of 1.2 mcg/mL. At this concentration, the extract was not cytotoxic to normally-dividing breast epithelial cells. Cytotoxicity of the extract was mediated via apoptosis as demonstrated by DNA fragmentation, caspase-3 activation and fluorescence microscopic analyses.
Conclusion
The study shows that the flavonoid-rich ethyl acetate fraction of the crude methanol leaf extract
of S. samarangense possesses potent apoptotic and cytotoxic properties against MCF-7 breast cancer cell lines at low concentrations.
MCF-7 Cells
;
Syzygium
3.Comparative Studies on the Polyarnine Involvement in MCF - 7 and MDA - MB - 231 Breast Cancer Cell Proliferation.
Journal of the Korean Cancer Association 1999;31(6):1151-1158
No abstract available.
Breast Neoplasms*
;
Breast*
;
Cell Proliferation*
;
MCF-7 Cells
;
Polyamines
4.Cytotoxic Lactones from the Pericarps of Litsea japonica
Quynh Mai Thi NGO ; Thao Quyen CAO ; Mi Hee WOO ; Byung Sun MIN
Natural Product Sciences 2019;25(1):23-27
From the pericarps of Litsea japonica (Thunb.) Jussieu, eighteen butanolide derivatives (1 – 18) were evaluated for their cytotoxic activity against HeLa, HL-60, and MCF-7 cells. Compounds 1 – 9 with 2-alkylidene-3-hydroxy-4-methylbutanolides structure exhibited cytotoxic activities against cancer-cell lines. Among them, compound 8 (litsenolide D₂) exhibited the most potent cytotoxicity against the tested cell lines, including HeLa, HL-60, and MCF-7, with IC₅₀ values of 17.6 ± 1.3, 4.2 ± 0.2, and 12.8 ± 0.0 µM, respectively. Compound 8 induced apoptosis in a dose-dependent manner. Annexin V/Propidium Iodide (PI) double staining confirmed that 8 effectively induced apoptosis in MCF-7 cells. To the best of our knowledge, we have reported cytotoxic activity of butanolides from L. japonica against these cancer-cell lines for the first time.
Apoptosis
;
Cell Line
;
Lactones
;
Lauraceae
;
Litsea
;
MCF-7 Cells
5.Comparison of cell elasticity analysis methods based on atomic force microscopy indentation.
Zhe WANG ; Fengtao HAO ; Xiaohu CHEN ; Zhouqi YANG ; Chong DING ; Peng SHANG
Journal of Biomedical Engineering 2014;31(5):1075-1079
In order to investigate in greater detail the two methods based on Hertz model for analyzing force-distance curve obtained by atomic force microscopy, we acquired the force-distance curves of Hela and MCF-7 cells by atomic force microscopy (AFM) indentation in this study. After the determination of contact point, Young's modulus in different indentation depth were calculated with two analysis methods of "two point" and "slope fitting". The results showed that the Young's modulus of Hela cell was higher than that of MCF-7 cell,which is in accordance with the F-actin distribution of the two types of cell. We found that the Young's modulus of the cells was decreased with increasing indentation depth and the curve trends by "slope fitting". This indicated that the "slope fitting" method could reduce the error caused by the miscalculation of contact point. The purpose of this study was to provide a guidance for researcher to choose an appropriate method for analyzing AFM indentation force-distance curve.
Actins
;
Elastic Modulus
;
HeLa Cells
;
cytology
;
Humans
;
MCF-7 Cells
;
cytology
;
Microscopy, Atomic Force
6.Reserpine treatment activates AMP activated protein kinase (AMPK).
Rackhyun PARK ; Kang Il LEE ; Hyunju KIM ; Minsu JANG ; Thi Kim Quy HA ; Won Keun OH ; Junsoo PARK
Natural Product Sciences 2017;23(3):157-161
Reserpine is a well-known medicine for the treatment of hypertension, however the role of reserpine in cell signaling is not fully understood. Here, we report that reserpine treatment induces the phosphorylation of AMP activated protein kinase (AMPK) at threonine 172 (T172) in PC12 cells. Phosphorylation of AMPK T172 is regulated by upstream signaling molecules, and the increase of phospho-T172 indicates that AMPK is activated. When we examined the FOXO3a dependent transcription by using the FHRE-Luc reporter assay, reserpine treatment repressed the FHRE-Luc reporter activity in a dose dependent manner. Finally, we showed that reserpine treatment induced the phosphorylation of AMPK as well as cell death in MCF-7 cells. These results suggest that AMPK is a potential cellular target of reserpine.
AMP-Activated Protein Kinases*
;
Animals
;
Cell Death
;
Hypertension
;
MCF-7 Cells
;
PC12 Cells
;
Phosphorylation
;
Reserpine*
;
Threonine
7.Apoptotic Effects of 6-Gingerol in Human Breast Cancer Cells.
Hyun Woo KIM ; Deuk Hee OH ; Jeong Tae KOH ; Young Chai LIM
International Journal of Oral Biology 2015;40(4):223-228
6-Gingerol exerts anti-tumor effects in various cancer cell models. We evaluated the effect of 6-gingerol on the growth of MCF-7 breast cancer cells and MCF-10A breast epithelial cells to determine whether any growth-inhibitory effects found were attributable to apoptosis, and to elucidate the underlying mechanism of action. 6-Gingerol inhibited the viability of both cell lines in a dose- and time-dependent manner; however, the degree of inhibition was greater in MCF-7 than MCF-10A cells. By flow cytometry, induction of dose- and time-dependent apoptosis was found, and the magnitude of apoptosis was also markedly greater in MCF-7 than MCF-10A cells. Expression of caspase-3 and poly (ADP-ribose) polymerase (PARP) was observed in MCF-7 cells treated with 6-gingerol, and further cleavage of PARP occurred in these cells. We suggest that 6-gingerol induces apoptosis in human breast cancer cells mainly by promoting caspase-3 expression and subsequent degradation of PARP.
Apoptosis
;
Breast Neoplasms*
;
Breast*
;
Caspase 3
;
Cell Line
;
Epithelial Cells
;
Flow Cytometry
;
Humans*
;
MCF-7 Cells
8.Three new diterpenoids from whole herb of Carpesium cernuum.
Shu-Hui FENG ; Wei-Qing ZHANG ; Wei LIANG ; Chen YAN
China Journal of Chinese Materia Medica 2023;48(19):5244-5249
The study investigated the chemical constituents from the whole herb of Carpesium cernuum. Three new diterpenoids were isolated from the whole herb of C. cernuum by column chromatography on silica gel, Sephadex LH-20, and semi-preparative HPLC. Their structures were identified by MS, NMR and other spectral techniques. The isolates were identified as(5Z)-2-oxo-2, 10, 14-trimethylhexadeca-5, 13-diene-11α, 18-diol(1),(2E, 10E)-7-[(acetyloxy)methyl]-3, 11, 15-trimethylhexadeca-2, 10, 14-triene-1, 12α-diol(2),(2E, 6Z)-3, 11, 15-trimethylhexadeca-2, 6, 14-triene-1, 12α, 19-triol(3), respectively. The cytotoxic activity of compounds 1-3 were investigated with DU-145, MCF-7, and A549 cells by MTT. The results showed that compound 1 and 3 had certain inhibitory effects on MCF-7 cells, with the inhibition rates of 45.06% and 29.40%, respectively.
Humans
;
Asteraceae/chemistry*
;
MCF-7 Cells
;
Magnetic Resonance Spectroscopy
;
Chromatography, High Pressure Liquid
;
A549 Cells
9.Cytotoxic, Anti-Inflammatory and Adipogenic Effects of Inophyllum D, Calanone, Isocordato-oblongic acid, and Morelloflavone on Cell Lines.
Muhammad TAHER ; Amnani AMINUDDIN ; Deny SUSANTI ; Nurul Iman AMINUDIN ; Shamsul ON ; Farediah AHMAD ; Hanisuhana HAMIDON
Natural Product Sciences 2016;22(2):122-128
This paper reports in vitro cytotoxic, anti-inflammatory and adipocyte diffentiation with adipogenic effects of coumarins inophyllum D (1) and calanone (2), and a chromanone carboxylic acid namely isocordato-oblongic acid (3) isolated from Calophyllum symingtonianum as well as a biflavonoid morelloflavon e (4) isolated from Garcinia prainiana on MCF-7 breast adenocarcinoma RAW 264.7 macrophages and 3T3-L1 preadipocytes cells, respectively. The cytotoxicity study on MCF-7 cell was conducted by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Meanwhile, the study of anti-inflammatory effects in RAW 264.7 macrophages and adipogenic effects on 3T3-L1 pre-adipocytes were conducted through nitrite determination assay and induction of adipocyte differentiation, respectively. In the cytotoxicity study, inophyllum D (1) was the only compounds that exhibited significant cytotoxic effect against MCF-7 cell with IC₅₀ of 84 µg/mL. Further, all by inhibiting the compounds have shown anti-inflammatory effects in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages of nitrite concentration with production. In addition, the compounds also exhibited adipogenic effects on 3T3-L1 pre-adipocytes by stimulating lipid formation. Thus, this study may provide significant input in discovery of the potential effects cytotoxic, anti-inflammatory and adipogenic agents.
Adenocarcinoma
;
Adipocytes
;
Breast
;
Calophyllum
;
Cell Line*
;
Coumarins
;
Garcinia
;
In Vitro Techniques
;
Macrophages
;
MCF-7 Cells
10.Transient Receptor Potential Vanilloid 1 Expression and Functionality in MCF-7 Cells: A Preliminary Investigation.
Cristina VERCELLI ; Raffaella BARBERO ; Barbara CUNIBERTI ; Silvia RACCA ; Giuliana ABBADESSA ; Francesca PICCIONE ; Giovanni RE
Journal of Breast Cancer 2014;17(4):332-338
PURPOSE: Transient receptor potential vanilloid 1 (TRPV1) is a nonselective cation channel belonging to the transient receptor potential family, and it is expressed in different neoplastic tissues. Its activation is associated with regulation of cancer growth and progression. The aim of this research was to study the expression and pharmacological characteristics of TRPV1 in cells derived from human breast cancer MCF-7 cells. METHODS: TRPV1 presence was assessed by binding studies and Western blotting. Receptor binding characteristics were evaluated through competition assays, while 3-(4,5-dimethylthiazol-2-yl)-2,5,-dipheyltetrazolium bromide reduction assays were performed to confirm an early hypothesis regarding the modulation of cancer cell proliferation. The functionality of TRPV1 was evaluated by measuring Ca2+ uptake in the presence of increasing concentrations of TRPV1 agonists and antagonists. RESULTS: Binding studies identified a single class of TRPV1 (Bmax 1,492+/-192 fmol/mg protein), and Western blot showed a signal at 100 kDa corresponding to the molecular weight of human TRPV1. Among the different tested agonists and antagonists, anandamide (Ki: 2.8x10(-11) M) and 5-iodoresiniferatoxin (5-I-RTX) (Ki: 5.6x10(-11) M) showed the highest degrees of affinity for TRPV1, respectively. All tested TRPV1 agonists and antagonists caused a significant (p<0.05) decrease in cell growth rate in MCF-7 cells. For agonists and antagonists, the efficacy of tested compounds displayed the following rank order: resiniferatoxin>anandamide>capsaicin and 5-I-RTX=capsazepine, respectively. CONCLUSION: These data indicate that both TRPV1 agonists and antagonists induce significant inhibition of MCF-7 cell growth. Even though the mechanisms involved in the antiproliferative effects of TRPV1 agonists and antagonists should be further investigated, it has been suggested that agonists cause desensitization of the receptor, leading to alteration in Ca2+-influx regulation. By contrast, antagonists cause a functional block of the receptor with consequent fatal dysregulation of cell homeostasis.
Blotting, Western
;
Breast Neoplasms
;
Cell Proliferation
;
Homeostasis
;
Humans
;
MCF-7 Cells*
;
Molecular Weight