Selective induction of DNA damage, G2 abrogation, and mitochondrial apoptosis by leaf extract of traditional medicinal plant Wrightia arborea in K562 cells

Abstract

Plants have proved to be an important source of anti-cancer drugs. Wrightia arborea, an Indian Ayurvedic medicinal plant, is used traditionally to treat a variety of ailments. This study evaluates the antiproliferative/apoptotic potential of Wrightia arborea leaf extracts, prepared in different organic solvents, on cancer cell lines. MTT assay, light and fluorescence microscopy, flow cytometry, DNA laddering, alkaline comet assay, and western blotting were some of the techniques used for evaluation. Combinations of camptothecin, either with CHK1 inhibitor-PD407824 or with W. arborea leaf extract, were deployed to determine the G2 abrogating potential of the extract. The chloroform extract (WAC) selectively killed K562 cells, without affecting cancerous MCF-7, Hep G2 cells, and normal human peripheral blood lymphocytes. Cell death was characterized by observation of apoptotic bodies, increased Ca2+ and ROS, phosphatidyl serine externalization, mitochondrial membrane depolarization, DNA laddering, increased sub-G1 population, and altered expression of caspase 3, -9, and PARP. WAC also induced DNA damage, alterations in key G2/M phase protein expression, cell cycle perturbation, and potent G2 abrogation. The present study showed that W. arborea leaf extract, WAC, is capable of selectively killing leukemic cancer cells leaving normal lymphocytes unaffected. Our results indicate that this is effectuated through DNA damage and G2 abrogation leading to mitochondrial apoptosis. Taken together, this report contributes toward a better understanding of the anticancer properties of this traditional medicinal plant extract possessing valuable bioactive constituents which can serve as a bioresource for promising complimentary/alternative/chemopreventive therapeutics.

GC-MS analysis and cytotoxic activity of the n-hexane fraction from Curcuma sahuynhensis Škornick. & N.S.Lý leaves collected in Vietnam

“…Where I: inhibition rate of cancer cells, ODSp: average optical density value of testing sample; ODBl: average optical density value of blank sample; ODDMSO: average optical density value of DMSO. HL60 cytotoxicity effect: The HL60 cytotoxicity effect of vegetable turmeric leaves n-hexane extract was conducted according to the method of Lakshmipriya et al with some minor modifications (17). The HL-60 test cell line was cultured in DMEM with the described above procedure.…”

Anticancer potential of rhizome extract and a labdane diterpenoid from Curcuma mutabilis plant endemic to Western Ghats of India

“…Cytotoxicity assay. For cytotoxicity evaluation, all cancer and normal cell types were exposed to varying concentrations (1-100 μg/mL) of different C. mutabilis rhizome extracts or column sub-fractions from CMRP, prepared in DMSO and incubated for 24 h, followed by MTT assay as previously described 47 .…”

“…Immunoblotting. SDS-PAGE and western blotting were performed essentially as described previously 47 .…”

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“…DNA Fragmentation Assay: DNA isolated from control and treated cells were analyzed on 1% agarose gels to check for DNA laddering arising due to internucleosomal cleavage, which is considered to be a hallmark of apoptosis induction 31 .…”