Cytotoxicity of Wortmannin Triggers Programmed Cell Death in MCF-7 Cells; Biochemical and (original) (raw)
Related papers
2016
The present study aimed to explore the molecular mechanisms and biological pathways involved in Wortmannin induced breast cancer MCF-7 programmed cell death. The direct cellular and molecular effect of Wtmn was investigated selectively on the MCF-7 cancer cell line. To study morphological effects phase contrast microscopy was used. The mitochondrial membrane potential and detection of caspase activity were investigated using fluorescent microscopy. Our morphological, molecular, and caspases expression indicated the intrinsic apoptosis pathways involved, and confirmed the role of Wtmn in these pathways. Our experimental results demonstrated that Wortmannin (Wtmn), an irreversible and selective PI3-K inhibitor, inhibits the proliferation of MCF-7 breast cancer cells, and facilitates their entry into apoptosis. Our data established the anti-cancer properties of Wtmn on MCF-7 breast cancer cells. These studies directed us towards elucidating the molecular mechanisms and biological pathw...
Breast Cancer: Targets and Therapy, 2012
Background: Apoptosis can be used as a reliable marker for evaluating potential chemotherapeutic agents. Because wortmannin is a microbial steroidal metabolite, it specifically inhibits the phosphatidyl inositol 3-kinase pathway, and could be used as a promising apoptosis-based therapeutic agent in the treatment of cancer. The objective of this study was to investigate the biomolecular mechanisms involved in wortmannin-induced cell death of breast cancer-derived MCF-7 cells. Methods and results: Our experimental results demonstrate that wortmannin has strong apoptotic effects through a combination of different actions, including reduction of cell viability in a dose-dependent manner, inhibition of proliferation, and enhanced generation of intracellular reactive oxygen species. Conclusion: Our findings suggest that wortmannin induces MCF-7 cell death via a programmed pathway showing chromatin condensation, nuclear fragmentation, reactive oxygen species, and membrane blebbing, which are characteristics typical of apoptosis.
Wogonin Stimulation of Cell Death and Reducing Survivin in MDM-MB231 Breast Tumors
ImmunoRegulation
Background: Wogonin as a flavonoid compound is known for its anticancer effect through growth control, differentiation, and apoptosis. This study investigates the possible activity of wogonin in inducing cell death of breast tumors MDA-MB231. Materials and Methods: The level of cell proliferation was evaluated by MTT assay. Cycle analysis and the apoptotic rate were assessed by flow cytometry. mRNA level of Bax, Bcl-2, p53, and survivin was evaluated by real-time PCR. Western blot was utilized to assert the relative protein expression. Results: Wogonin inhibits the proliferation of MDA-MB231 over time, and in particular, wogonin can induce the arrest of phase G1 of MDA-MB231 cells. The apoptosis of wogonin was related to a remarkable decrease in Bcl-2 and survivin and an increment in p53 and Bax. Wogonin also significantly elevated the active apoptotic forms of caspase-3, -8, -9. Z-DEVD-FMK, a specific inhibitor of caspase-3, remarkably inhibits cellular apoptosis caused by wogonin....
Apoptotic Effect of Wortmannolone on Cancer Cells through Potent ROS Induction
International journal of cancer research, 2013
Nuclear factor κappa-B inhibitors isolated from natural sources that induce apoptosis are promising new agents with anticancer properties. Wortmannin and wortmannolone were isolated from endophytic fungus (Penicillum polonicum) and showed NF-κB inhibitory effects with inhibitory concentration (IC50) of 0.47 μM and 2.06 μM for wortmannin and wortmannolone, respectively. The activity was compared with rocaglamide (IC50=0.075 μM). The mechanism through which wortmannin and wortmannolone exhibited an attenuating effect on the NF-κB pathway was further evaluated in this study. Wortmannolone showed significant reactive oxygen species (ROS) inducing effects in HeLa cervical cells. The ROS inducing effect was concentration dependent, and the ROS generating activity was comparable with daunomycin, a potent chemotherapeutic agent. The findings suggested that the elevated formation of ROS was partially involved in the induction of apoptosis in treated cells. Potent cytotoxic and apoptotic effe...
Wogonin sensitizes resistant malignant cells to TNFα- and TRAIL-induced apoptosis
2006
TNFα has previously been used in anti-cancer therapy. However, the therapeutic application of TNFα was largely limited due to its general toxicity and the fact that it activates the NF-κB-family transcription factors which are pro-inflammatory and antiapoptotic. To overcome this problem in vitro, specific NF-κB inhibitors, transcription or protein synthesis inhibitors such as actinomycin D and cycloheximide are usually used in combination to increase TNFα-killing of tumor cells. However, these agents also cause harmful side effects in vivo. We show here that wogonin, derived from the popular Chinese herb Huang-Qin, attenuates NF-κB activity by shifting TNFα-induced free radical . O 2 to a more reduced non-radical product H 2 O 2 and thereby sensitizes TNFα-resistant leukemia cells to TNFα-induced apoptosis. Importantly, wogonin does not affect the viability of normal peripheral blood T cells. Wogonin also sensitizes TRAIL-induced apoptosis. Our data suggest a potential use of wogonin as a TNFα or TRAIL adjuvant for cancer treatment. Our data also demonstrate how an herbal compound enhances killing of tumor cells at reduced side effects compared to other treatments.
Differential Effect of Wortmannolone Derivatives on MDA-MB-231 Breast Cancer Cells
Anticancer research, 2017
The survival rate of women diagnosed with triple-negative breast-cancer (TNBC) remains low. Hence, this study aimed at the chemical and biological optimization of furanosteroid derivatives for the treatment of this type of malignancy using TNBC cells. Semi-synthetic analogs of wortmannolone (1-6) that negatively affected the aberrant pathways in tumor cells were evaluated in hormone-independent breast cancer cells using western blot and cell-cycle analysis. Wortmannolone derivatization generated NF-ĸB inhibitors as new lead structures for further development. Compound (3) was found to be the most significantly active lead. Structure-activity analysis in the present study showed that acetylation of the hydroxyl groups and substitution on C3 and C17 of wortmannolone enhanced biological activity. Alpha-substitution of the acetyl group in C3 on ring A (compound 3) resulted in ROS inducing effect; however, presence of an acetyl group in β-position of C3 displayed the highest NF-ĸB p65 in...
Biochemical Pharmacology, 2002
Seven structurally related¯avonoids including luteolin, nobiletin, wogonin, baicalein, apigenin, myricetin and ®setin were used to study their biological activities on the human leukemia cell line, HL-60. On MTT assay, wogonin, baicalein, apigenin, myricetin and ®setin showed obvious cytotoxic effects on HL-60 cells, with wogonin and ®setin being the most-potent apoptotic inducers among them. The cytotoxic effects of wogonin and ®setin were accompanied by the dose-and time-dependent appearance of characteristics of apoptosis including DNA fragmentation, apoptotic bodies and the sub-G1 ratio. Treatment with an apoptosis-inducing concentration of wogonin or ®setin causes rapid and transient induction of caspase 3/CPP32 activity, but not caspase 1 activity. Further, cleavage of poly(ADP-ribose) polymerase (PARP) and decrease of pro-caspase 3 protein were detected in wogonin-and ®setin-treated HL-60 cells. An increase in the pro-apoptotic protein, bax, and a decrease in the anti-apoptotic protein, Mcl-1, were detected in ®setin-and wogonintreated HL-60 cells. However, Bcl-2, Bcl-XL, and Bad all remained unchanged in wogonin-and ®setin-treated HL-60 cells. In vitro chromatin digestion revealed that endonuclease activity was profoundly enhanced in wogonin-and ®setin-treated HL-60 cells, and the addition of ethylenediaminetetraacetic acid (EDTA) or ethyleneglycoltetraacetic acid (EGTA) into the reaction blocked endonuclease activation and at an optimum pH of 7.5. The caspase 3 inhibitor, Ac-DEVD-CHO, but not the caspase 1 inhibitor, Ac-YVAD-CHO, attenuated wogonin-and ®setin-induced DNA ladders, PARP cleavage, and endonuclease activation. Pretreatment of HL-60 cells with N-acetyl-cysteine or catalase ef®ciently inhibited H 2 O 2 (200 mM)-induced apoptosis, but showed no inhibitory effect on wogonin-and ®setin-induced DNA ladders, caspase 3 activation, or bax protein induction. Decrease in endogenous ROS production was detected in wogonin-and ®setin-treated HL-60 cells by DCHF-DA assay. In conclusion, our experiments indicate that a decrease in intracellular peroxide level was involved in wogonin-and ®setin-induced apoptosis; activation of caspase 3 and endonuclease, induction of bax protein and suppression of Mcl-1 protein were detected in the process.
Journal of Cellular Biochemistry, 2008
We investigate the roles of methoxyl (OCH 3) and hydroxyl (OH) substitutions at C8 of flavonoids on their apoptosis-inducing activities. Wogonin (Wog) and nor-wogonin (N-Wog) are structurally related flavonoids, and respectively contain an OH and OCH 3 at C8. In leukemia HL-60 cells, N-Wog exhibited more-potent cytotoxicity than Wog according to the MTT and LDH release assays, and the IC 50 values of Wog and N-Wog in HL-60 cells were 67.5 AE 2.1 and 21.7 AE 1.5 mM, respectively. Apoptotic characteristics including DNA ladders, apoptotic bodies, and hypodiploid cells accompanied by the induction of caspase 3 protein processing appeared in Wog-and N-Wog-treated HL-60 cells. Interestingly, an increase in intracellular peroxide production was detected in N-Wog-but not Wog-treated HL-60 cells by the DCHF-DA assay, and the reduction of intracellular peroxide by catalase (CAT) induced by N-Wog significantly reduced the N-Wog-but not the Wog-induced cytotoxic effect according to the MTT assay in accordance with the blocking of DNA ladder formation and caspase 3 and PARP protein processing elicited by N-Wog. We further analyzed the effect of six structurally related compounds, including 5-OH, 7-OH, 5,7-diOH, 5,7-diOCH 3 , 7,8-diOCH 3 , and 7-OCH 3-8-OH flavones, on apoptosis induction in HL-60 cells. Results suggested that OH at C5 and C7 is essential for both the apoptosis-inducing activity of flavonoids, and OH at C8 may contribute to apoptosis induction ability. Evidence to support a distinct structure-activity relationship in apoptosis induction of flavonoids is provided for the first time in this study.
Intracellular effects of prodrug-like wortmannin probes
Chinese Chemical Letters, 2018
Wortmannin, a known inhibitor of phosphoinositide 3-kinases (PI3Ks), their low selectivity and high toxicity is still problematic and less is known about their effects on PI3Ks in cellular systems. Hence, we have synthesized a series of multifunctional wortmannin probes with the ability to self-activate, by installing a clickable handle at C11 site, and secondary amine and cancer-targeting moiety at C20 site, respectively. MTT assay first confirmed that self-activating probes have better inhibition potency and biotin enhanced their cancer cell uptake. Further experiments showed most of probes can target PI3K/ Akt/mTOR pathway with prolonged turnover time. Protein profiling and pull-down results were observed that the derivatives not only labelled four PI3Ks with selectivity, but also engaged in covalent interactions with numerous cellular proteins which could be the major reason of their high toxicity.
Regulation of cell signaling pathways by Wogonin in different cancers: Mechanistic review
Cellular and Molecular Biology, 2021
Natural products have historically been invaluable as a premium source of therapeutic agents. Recent advancements in genomics and structural biology have portrayed a high-resolution landscape of the diversity of proteins targeted by pharmacologically active products from natural sources. Natural product research has generated valuable wealth of information and cutting-edge research-works have leveraged our conceptual knowledge altogether to a new level. Wogonin (5,7-dihydroxy-8-methoxyflavone) is an O-methylated flavone and has attracted noteworthy appreciation because of its ability to pharmacologically target plethora of cell signaling pathways in different cancers. In this mini-review, we have gathered scattered pieces of available scientific evidence to summarize how wogonin pharmaceutically targeted Wnt/β-catenin, JAK/STAT, VEGF/VEGFR and TRAIL-driven apoptotic pathways in wide variety of cancers. We have also critically analyzed how wogonin prevented carcinogenesis and metasta...