Apoptotic and Autophagic Effects of Sesbania grandiflora Flowers in Human Leukemic Cells

Abstract

BackgroundIdentification of cytotoxic compounds that induce apoptosis has been the mainstay of anti-cancer therapeutics for several decades. In recent years, focus has shifted to inducing multiple modes of cell death coupled with reduced systemic toxicity. The plant Sesbania grandiflora is widely used in Indian traditional medicine for the treatment of a broad spectrum of diseases. This encouraged us to investigate into the anti-proliferative effect of a fraction (F2) isolated from S. grandiflora flowers in cancer cells and delineate the underlying involvement of apoptotic and autophagic pathways.Principal FindingsUsing MTT based cell viability assay, we evaluated the cytotoxic potential of fraction F2. It was the most effective on U937 cells (IC50∶18.6 µg/ml). Inhibition of growth involved enhancement of Annexin V positivity. This was associated with elevated reactive oxygen species generation, measured by flow cytometry and reduced oxygen consumption – both effects being abrogated by anti-oxidant NAC. This caused stimulation of pro-apoptotic proteins and concomitant inhibition of anti-apoptotic protein expressions inducing mitochondrial depolarization, as measured by flow cytometry and release of cytochrome c. Interestingly, even with these molecular features of apoptosis, F2 was able to alter Atg protein levels and induce LC3 processing. This was accompanied by formation of autophagic vacuoles as revealed by fluorescence and transmission electron microscopy – confirming the occurrence of autophagy. Eventually, F2 triggered caspase cascade – executioners of programmed cell death and AIF translocation to nuclei. This culminated in cleavage of the DNA repair enzyme, poly (ADP-ribose) polymerase that caused DNA damage as proved by staining with Hoechst 33258 leading to cell death.ConclusionsThe findings suggest fraction F2 triggers pro-oxidant activity and mediates its cytotoxicity in leukemic cells via apoptosis and autophagy. Thus, it merits consideration and further investigation as a therapeutic option for the treatment of leukemia.

Should charge variants of monoclonal antibody therapeutics be considered critical quality attributes?

“…B and it is seen that the model is statistically relevant ( R 2 = 0.85, RMSE = 5.75) (Fig. B) . It is expected that the main product would exhibit antiproliferation on the fibroblast cells and this is consistent with the prediction of the model.…”

“…However, not all of these variants impacted the proliferation activity of the cells. Therefore, the variants exhibiting significant effect based on the abundance, sensitivity in terms of their ability to cause change in proliferative activity per unit change in percentage variant, or both were selected to build a refined model as per the procedure outlined in . In the refined model, the significance of the parameters was determined based on p ‐values ( p < 0.0001) and F value (ratio of mean square of the model to the mean square of the error).…”

Antiproliferative and apoptosis inducing effect of essential oil extracted from Cyrtomium fortumei (J.) Smith leaves

“…The inhibitory effect of EO on cancer cells via induction of apoptosis was further confirmed by Hoechst 33258 staining. Live cells with homogeneous light blue nuclei could be observed under fluorescence microscopy after staining with Hoechst 33258, and the apoptotic cells displayed bright blue due to karyopyknosis and chromatin condensation after treating with Hoechst 33258, and the nuclei of dead cells could not be stained (Roy et al, 2013).…”

Medicinal and Therapeutic Value of Sesbania grandiflora

“…Plants contain a huge range of active compounds with the most abundant being polyphenols, carotenoids, vitamin (vitamin A, C, riboflavin, nicotinic acid), and minerals like zinc and selenium which form an integral part of antioxidant systems and reduce cellular damages [21,22]. Roy et.al, 2013 reported that a fraction isolated from flowers, preferentially kills leukemic cells (particularly those of histiocyte lymphoma) by triggering programmed cell death [23]. In 4 different studies from 2012 to 2016, it was found that the flower, fruit and the whole plant extract reduced blood glucose, cholesterol, TG and LDL, lipid peroxidation and increased and superoxide dismutase and catalase of insulin and hemoglobin in experimental animals [24][25][26][27].…”