title: Synergistic effect of piperine and paclitaxel on cell fate via cyt-c, Bax/Bcl-2-caspase-3 pathway in ovarian adenocarcinomas SKOV-3 cells.
abstract: Ovarian cancer is fourth most common and lethal among all gynecologic malignancies. The chemotherapy usually requires in all stages of ovarian cancer but drugs have several side effects. We hypothesized that use of combination therapy of paclitaxel (PTX) and phytochemical piperine (PIP) may reduce the PTX dose as well as toxicity. The human ovarian adenocarcinomas SKOV3 cell treated with PTX-5nM and PIP-10µm after determination of IC50by MTT assay. Reactive oxygen species generation, mitochondrial membrane potential (MMP), DNA damage, cell death pathway markers as release of cyt-c, Bax/Bcl2-caspase-3 and cell cycle arrest were analyzed. The dose dependent treatment of SKOV-3 cells showed IC50and synergism at combination of 5nM-PTX and 10µm-PIP in cell viability assay. PTX and PIP increases the accumulation of reactive oxygen species which subsequently leading to increase in JC-1 and fragmented nuclei in mitotracker/DAPI staining. Comet assay showed 4.4-fold increase of tail formation in combined treated cells as compared to control. PTX-PIP arrests the cell cycle in sub-G1 phase. Immunocytochemistry of Bax showed increase in red fluorescence intensity whereas decrease in green fluorescence i.e Bax/Bcl-2 ratio increased. Moreover morphological EB/AO and Hoechst staining confirmed the enhanced apoptosis in combined treatment. Significant upregulation of apoptotic genes, cyt-c (3.4 fold) Bax (2.8 fold), caspase-3 (3.6 fold) whereas no change occurred in Bcl2 mRNA expression and protein expressions. The combination of PTX with PIP produces synergistic effects in SKOV-3 cells via the modulation of pro and anti-apoptotic gene and may compensate the toxicity and side effects of PTX.
authors.affiliation: Department of Obstetrics and Gynecology, KGMU, Lucknow 226003, India; Photobiology Division, CSIR-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Manish Kumar
authors.initials: MK
authors.lastname: Pal
authors.affiliation: Department of Obstetrics and Gynecology, KGMU, Lucknow 226003, India. Electronic address: spjaiswar59@gmail.com.
authors.forename: Shyama Pyari
authors.initials: SP
authors.lastname: Jaiswar
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Ajeet Kumar
authors.initials: AK
authors.lastname: Srivastav
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Shruti
authors.initials: S
authors.lastname: Goyal
authors.affiliation: Department of Zoology, Banaras Hindu University, Varanasi, India.
authors.forename: Ashish
authors.initials: A
authors.lastname: Dwivedi
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Ankit
authors.initials: A
authors.lastname: Verma
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow 226001, India.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Singh
authors.affiliation: Dr. Ram Manohar Lohia Institute of Medical Sciences, Lucknow, India.
authors.forename: Anumesh Kumar
authors.initials: AK
authors.lastname: Pathak
authors.affiliation: Department of Obstetrics and Gynecology, KGMU, Lucknow 226003, India. Electronic address: sankhwar_pushpa@yahoo.com.
authors.forename: Pushpa Lata
authors.initials: PL
authors.lastname: Sankhwar
authors.affiliation: Photobiology Division, CSIR-Indian Institute of Toxicology Research, Post Box No. 80, M.G. Marg, Lucknow 226001, India. Electronic address: ratanray.2011@rediffmail.com.
authors.forename: Ratan Singh
authors.initials: RS
authors.lastname: Ray
mesh_terms: D000230:Adenocarcinoma / Q000473:pathology*; D000470:Alkaloids / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D052117:Benzodioxoles / Q000494:pharmacology*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D004357:Drug Synergism; D005260:Female; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D051336:Mitochondrial Membranes / Q000187:drug effects; D010051:Ovarian Neoplasms / Q000473:pathology*; D017239:Paclitaxel / Q000494:pharmacology*; D010880:Piperidines / Q000494:pharmacology*; D053284:Polyunsaturated Alkamides / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D000470:Alkaloids; D000970:Antineoplastic Agents; D052117:Benzodioxoles; D010880:Piperidines; D053284:Polyunsaturated Alkamides; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; D053148:Caspase 3; D017239:Paclitaxel; C008922:piperine
keywords: Apoptosis; Caspase-3; DNA damage; Paclitaxel; Piperine