title: A novel alkaloid, evodiamine causes nuclear localization of cytochrome-c and induces apoptosis independent of p53 in human lung cancer cells.
abstract: Lung cancer is the most frequently diagnosed malignancy that contributes to high proportion of deaths globally among patients who die due to cancer. Chemotherapy remains the common mode of treatment for lung cancer patients though with limited success. We assessed the biological effects and associated molecular changes of evodiamine, a plant alkaloid, on human lung cancer A549 and H1299 cells along with other epithelial cancer and normal lung SAEC cells. Our data showed that 20-40 μM evodiamine treatment for 24-48 h strongly (up to 73%, P < 0.001) reduced the growth and survival of these cancer cells. However, it also moderately inhibited growth and survival of SAEC cells. A strong inhibition (P < 0.001) was observed on clonogenicity of A549 cells. Further, evodiamine increased (4-fold) mitochondrial membrane depolarization with 6-fold increase in apoptosis and a slight increase in Bax/Bcl-2 ratio. It increased the cytochrome-c release from mitochondria into the cytosol as well as nucleus. Cytosolic cytochrome-c activated cascade of caspase-9 and caspase-3 intrinsic pathway, however, DR5 and caspase-8 extrinsic pathway was also activated which could be due to nuclear cytochrome-c. Pan-caspase inhibitor (z-VAD.fmk) partially reversed evodiamine induced apoptosis. An increase in p53 as well as its serine 15 phosphorylation was also observed. Pifithrin-α, a p53 inhibitor, slightly inhibited growth of A549 cells and under p53 inhibitory condition evodiamine-induced apoptosis could not be reversed. Together these findings suggest that evodiamine is a strong inducer of apoptosis in lung epithelial cancer cells independent of their p53 status and that could involve both intrinsic as well as extrinsic pathway of apoptosis. Thus evodiamine could be a potential anticancer agent against lung cancer.
authors.affiliation: School of Life Sciences, Central University of Gujarat, Gandhinagar, Gujarat, India.
authors.forename: Vijay
authors.initials: V
authors.lastname: Mohan
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Colorado Denver, Aurora, CO, USA.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Agarwal
authors.affiliation: School of Life Sciences, Central University of Gujarat, Gandhinagar, Gujarat, India; Cancer Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi, India. Electronic address: ranaps@hotmail.com.
authors.forename: Rana P
authors.initials: RP
authors.lastname: Singh
mesh_terms: D000072283:A549 Cells; D000470:Alkaloids / Q000008:administration & dosage; D000970:Antineoplastic Agents / Q000008:administration & dosage; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism* / Q000473:pathology; D019913:Nuclear Localization Signals / Q000187:drug effects; D011799:Quinazolines / Q000008:administration & dosage*; D014018:Tissue Distribution; D016896:Treatment Outcome; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D000470:Alkaloids; D000970:Antineoplastic Agents; D019913:Nuclear Localization Signals; D011799:Quinazolines; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D045304:Cytochromes c; C049639:evodiamine
keywords: Apoptosis; Cytochrome-c; Evodiamine; Lung cancer; Pifithrin-α; p53