scholarly journals Apoptosis Induction of Agave lechuguilla Torrey Extract on Human Lung Adenocarcinoma Cells (SK-LU-1)

2018 ◽  
Vol 19 (12) ◽  
pp. 3765 ◽  
Author(s):  
Luis Anguiano-Sevilla ◽  
Eugenia Lugo-Cervantes ◽  
Cynthia Ordaz-Pichardo ◽  
Jorge Rosas-Trigueros ◽  
María Jaramillo-Flores
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Ethanol Extract ◽  
Fragmentation Pattern ◽  
Intrinsic Pathway ◽  
Adenocarcinoma Cells ◽  
Human Lung Adenocarcinoma Cells ◽  
Caspase 7 ◽  
Mcf 7 ◽  
Apoptotic Signal

In this study, an ethanol extract of Agave lechuguilla was evaluated against six carcinogenic cell lines (HCT-15, MCF-7, PC-3, U-251, SK-LU-1 and K-562) with an inhibition of 75.7 ± 2.3% against the SK-LU-1 line. Based on the previous result, the extract was hydrolyzed and fractionated, to which the IC50 was determined; the cell line was more sensitive to the fractionated extract with an IC50 6.96 ± 0.15 µg/mL. Characterization by mass spectrometry showed the presence of kaempferol, quercetin and a flavonoid dimer formed by afzelechin-4β-8-quercetin, according to the generated fragmentation pattern. The fractionated extract presented cell death by apoptosis with 39.8% at 24 h. Molecular docking was performed with the molecules found to try to describe cell death by apoptosis through death receptors such as FasCD95, TNF-R1, DR4/5 and blocking signaling on the EGFR and K-Ras MAPK/ERK pathway, as well as through the intrinsic pathway activating tBID, which promotes the amplification of the apoptotic signal due to the activation of caspase-3, and consequently caspase-7. In addition to the activation of the IIb complex associated with cell death due to necroptosis.

The Cytotoxic Effect of Ethanol Extract of Momordica Charantia, Kuguacin-J and Cisplatin on Healthy MCF-10A and MCF-7 and MDAMB-231 Breast Cancer Cell Lines in Vitro

2021 ◽  
Author(s):  
Chahinez Houacine ◽  
Jaipaul Singh ◽  
Raphael Singh ◽  
Karishma Jeeboo ◽  
Abdullah Adil Ansari ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Death ◽  
Cell Viability ◽  
Cell Lines ◽  
Caspase 3 ◽  
Ethanol Extract ◽  
Momordica Charantia ◽  
High Dose ◽  
Time Points ◽  
Mcf 7

Abstract Traditional medicines, derived from plants, could present alternative treatment strategy for cancer therapy. One such plant is Momordica charantia (MC) which possesses anti-carcinogenic properties. This study investigated the anticancer effect of an ethanol extract of MC fruit, Kuguacin-J (K-J), an isolated compound from the leaves of MC and cisplatin, either alone or combination on healthy MCF-10A mammary cells and compared with breast cancer MCF-7 and MDAMB-231 cell lines. Cell viability was tested using 8 μg/mL and 80 μg/mL doses of MC extract, K-J and cisplatin individually or combined for 24 and 48 hours. Caspase-3- activity was measured in MCF-7 and MDA-MB-231 cells using established methods. The results revealed that MC extract and K-J had no effect on healthy MCF-10A cell viability as compared to cisplatin which induced dose and time-dependent cell death. Similarly, treatment of MCF-7 cells with cisplatin induced cell death at high concentration at both the time points, while MC extract and K-J only induce MCF-7 cell death at high dose after 48 hours only. During combination therapy, both doses of cisplatin enhanced MCF-7 cell death when combined with MC extract or K-J after 24 and 48 hours. In MDAMB-231 cells, the three drugs, either alone or combined, evoked significant cell death at both the doses and time points. All three drugs, at high dose, elicited significant increase in caspase-3- activity in MCF-7 and MDA-MB-231 cells as compared to untreated cells. The results revealed that either MC extract or K-J alone or combined with cisplatin, can elicit significant increase in cell death and caspase–3-activity in MCF-7 and MDA-MB-231cells as compared to untreated cells.

scholarly journals A novel method to detect intracellular metabolite alterations in MCF-7 cells by doxorubicin induced cell death

2019 ◽  
Author(s):  
Ajay Kumar ◽  
Sheetal Patel ◽  
Devyani Bhatkar ◽  
Nilesh Kumar Sharma
Keyword(s):  
Cell Death ◽  
Cancer Cells ◽  
Gel Electrophoresis ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Vertical Tube ◽  
Metabolic Reprogramming ◽  
Intracellular Metabolite ◽  
Intracellular Metabolites ◽  
Mcf 7

ABSTRACTMetabolic reprogramming within cancer cells is suggested as a potential barrier to chemotherapy. Additionally, metabolic tumor heterogeneity is one of factor behind discernible hallmarks such as drug resistance, relapse of tumor and the formation of secondary tumors. In this paper, cell based assays including PI/annexin V staining and immunoblot assay were performed to show the apoptotic cell death in MCF-7 cells treated with DOX. Further, MCF-7 cells were lysed in hypotonic buffer and whole cell lysate was purified by a novel and specifically designed metabolite (100 to 1000 Da) fractionation system as vertical tube gel electrophoresis (VTGE). Further, purified intracellular metabolites were subjected to identification by LC-HRMS technique. The authors show the presence of cleaved PARP 1 in MCF-7 cells treated with DOX. Concomitantly, data show the absence of active caspase 3 in MCF-7 cells. Novel findings are to identify key intracellular metabolites assisted by VTGE system that include lipid (CDP-DG, phytosphingosine, dodecanamide), non-lipid (N-acetyl-D-glucosamine, N1-acetylspermidine and gamma-L-glutamyl-L-cysteine) and tripeptide metabolites in MCF-7 cells treated by DOX. Interestingly, the authors report a first evidence of doxorubicinone, an aglycone form of DOX in MCF-7 cells that is potentially linked to the mechanism of cell death in MCF-7 cells. This paper reports on novel methods and processes that involve VTGE system based purification of hypotonically lysed novel intracellular metabolites of MCF-7 cells treated by DOX. Here, these identified intracellular metabolites corroborate to caspase 3 independent and mitochondria induced apoptotic cell death in MCF-7 cells.SIGNIFICANCE STATEMENTMetabolic reprogramming in cancer cells is implicated in various tumor hallmarks. Interestingly, thousands of research have addressed the molecular basis of drug treatment and resistance in chemotherapy. But, there is a significant gap in the precise methodologies and approaches in addressing intracellular metabolite alterations. This paper reports on a novel approach that helped reveal new findings on intracellular metabolite changes in case of doxorubicin (DOX) induced cell death in MCF-7 cells. This paper highlights the additional insights on debatable findings available in literature in the contexts of DOX induced cell death mechanisms. In this paper, novel and specifically designed vertical tube gel electrophoresis (VTGE) system is claimed to purify intracellular metabolites and this method is compatible with other biological system.

scholarly journals The ethanol extract of Garcinia subelliptica Merr. induces autophagy

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Kyun Ha Kim ◽  
Ji Yeon Lee ◽  
Wan Yi Li ◽  
Sangwoo Lee ◽  
Han-Sol Jeong ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Human Lung ◽  
Ethanol Extract ◽  
Cell Types ◽  
Hek293 Cells ◽  
Human Lung Adenocarcinoma ◽  
Ampk Pathway ◽  
Adenocarcinoma Cells ◽  
Human Lung Adenocarcinoma Cells ◽  
Cancer Suppression

Abstract Background Garcinia subelliptica Merr. is a multipurpose coastal tree, the potential medicinal effects of which have been studied, including cancer suppression. Here, we present evidence that the ethanol extract of G. subelliptica Merr. (eGSM) induces autophagy in human lung adenocarcinoma cells. Methods Two different human lung adenocarcinoma cell lines, A549 and SNU2292, were treated with varying amounts of eGSM. Cytotoxicity elicited by eGSM was assessed by MTT assay and PARP degradation. Autophagy in A549 and SNU2292 was determined by western blotting for AMPK, mTOR, ULK1, and LC3. Genetic deletion of AMPKα in HEK293 cells was carried out by CRISPR. Results eGSM elicited cytotoxicity, but not apoptosis, in A549 and SNU2292 cells. eGSM increased LC3-II production in both A549 and, more extensively, SNU2292, suggesting that eGSM induces autophagy. In A549, eGSM activated AMPK, an essential autophagy activator, but not suppressed mTOR, an autophagy blocker, suggesting that eGSM induces autophagy by primarily activating the AMPK pathway in A549. By contrast, eGSM suppressed mTOR activity without activating AMPK in SNU2292, suggesting that eGSM induces autophagy by mainly suppressing mTOR in SNU2292. In HEK293 cells lacking AMPKα expression, eGSM increased LC3-II production, confirming that the autophagy induced by eGSM can occur without the AMPK pathway. Conclusion Our findings suggest that eGSM induces autophagy by activating AMPK or suppressing mTOR pathways, depending on cell types.

A Newly Developed IAP Inhibitor (IAPi) Induces Apoptosis of Human Myeloma Cells and Synergises with Conventional and Novel Anti-Myeloma Therapeutics

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 5156-5156
Author(s):  
Tiffany T. Khong ◽  
Andrew Spencer
Keyword(s):  
Bone Marrow ◽  
Cell Death ◽  
Caspase 3 ◽  
Mononuclear Cells ◽  
Nuclear Translocation ◽  
Receptor Activation ◽  
Cell Killing ◽  
Parp Cleavage ◽  
Dependent Manner ◽  
Caspase 7

Abstract The IAPs (inhibitor of apoptosis proteins) are a family of 8 structurally related caspase inhibitors that efficiently block the execution phase of apoptosis induced by various stimuli including death receptor activation. Based on reports of elevated expression of IAPs in multiple myeloma (MM) we evaluated the therapeutic potential of a novel IAP (IAPi) in MM. A panel of 9 genetically heterogeneous human myeloma cell lines (HMCL) were screened and found to be sensitive to IAPi with an IC50 of 25 to 50μM in a dose and time dependent manner. Induction of cell death was confirmed by Annexin V and propidium iodide staining of treated HMCL. Mechanistic studies were then undertaken utilising 3 HMCL with high (NCI H929), intermediate (LP-1) or low (OPM2) sensitivity to IAPi with QRT-PCR demonstrating the highest expression of XIAP in the most IAPi sensitive HMCL NCI H929. In all 3 HMCL IAPi exposure was associated with rapid cleavage of caspase 3, caspase 7, ROCK-1, cytosolic degradation of ICAD, nuclear translocation of CAD and PARP cleavage confirming cell death via apoptosis. Replicate experiments utilising caspase specific inhibitors and siRNA knock-down of caspases 3 and 7 confirmed this to be both caspase 3 and caspase 7 dependent. Co-culture of HMCL with conditioned media from the human stromal cell line HS-5 or primary MM bone marrow mononuclear cells reproducibly demonstrated enhanced IAPi-induced HMCL apoptosis with a median increase in HMCL killing of 1.2 fold compared to HMCL treated in standard culture media. Similarly, IAPi activity was not abrogated but enhanced in the presence of either IL-6 or IGF-1 (1.1 and 1.3 fold, respectively) when compared to HMCL treated without the addition of exogenous growth factors. Primary MM cells from 9 multiply relapsed MM patients were treated with IAPi in an autologous bone marrow co-culture assay with concentrations of IAPi ranging from 10–100μM inducing a median of 23% (range, 8–56% as determined by Apo2.7 staining) MM cell killing at 48 hours. Finally the anti-MM activity of IAPi in combination with other conventional (etoposide, adriamycin, velcade) and novel (HSP90 inhibitor, agonistic TRAIL DR5 monoclonal antibody) therapeutics was investigated against both HMCL and primary MM tumour samples. All combinations showed synergistic cell killing with Combination Indices of <1 (Calcusyn) when compared to either agent alone. We conclude that IAPi induces apoptosis of MM cells via a caspase 3 and caspase 7 dependent process. Furthermore, IAPi retains anti-MM activity in the context of pro-survival cytokine exposure and induces synergistic killing of MM cells when combined with both conventional and novel anti-MM therapeutics. Based on these preliminary observations further investigation of the role of IAPi as a potential tumour sensitising agent for the therapy of MM is justified.

Sign in / Sign up

Export Citation Format

Share Document