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Low‐Dose Aspirin Down‐regulates the Expression of Transcription Factors and Anti‐apoptotic Genes in Human Cervical Cancer Cells
Author(s) -
Rasheed Huma,
Channa Iffat S,
Alvi Ayesha T,
Abbasi Atiya
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.lb97
Subject(s) - apoptosis , dna fragmentation , survivin , aspirin , cytotoxic t cell , hela , microbiology and biotechnology , fragmentation (computing) , cancer cell , chemistry , trypan blue , biology , cyclooxygenase , cancer research , programmed cell death , cancer , cell , enzyme , biochemistry , genetics , ecology , in vitro
Aspirin belongs to a class of Non‐Steroidal Anti‐Inflammatory Drugs (NSAIDs) that reduce pain and inflammation by the inhibition of an enzyme, cyclooxygenase (COX). The COX‐2 isoform of the enzyme is reported to regulate anti‐apoptotic genes in cancer of various origin such as colorectal, ovarian and cervical cancer. The present study deals with the investigation of the anti‐cancer effect of low dose aspirin and its underlying mechanisms in human cervical cancer (HeLa) cells. Cells were cultured in a sterile condition in DMEM with 10% FBS and 5% CO 2 . The cells were then treated with various concentrations (75 μM, 100 μM & 200 μM) of aspirin and used in different assays. Anti‐proliferative and cytotoxic effects of aspirin were studied by trypan blue exclusion assay, cell titer blue (CTB) and colony formation assays. To determine the molecular mechanism involved in the anti‐proliferative effect of aspirin the morphological and apoptotic changes were observed using fluorescence microscopy whereas nuclear degradation was assessed via DNA fragmentation. Finally, the Real Time‐qPCR was performed to detect the mRNA expression levels of cyclooxygenases (COX‐1 & COX‐2), transcription factors (c‐Myc, c‐Jun, c‐Fos) and genes involved in apoptotic pathways (Bcl2, Survivin). The results indicate that aspirin showed potent anti‐proliferative and cytotoxic effect in a concentration dependent manner ranged from 75 to 200 μM as well as inhibited the colony formation of Hela cells. The chromatin condensation and early apoptotic changes were found at 75μM and further a DNA fragmentation was visualized at 200 μM whereas the untreated controlled cells were remained intact. The expression of COX‐1, BCl2, surviving, c‐fos and c‐Jun mRNA was completely inhibited at 75μM of aspirin whereas c‐Myc and COX‐2 were inhibited at 200 μM. Our results demonstrate that aspirin exerts a potent antitumor effect on human cervical cancer cells through the suppression of transcription factors and anti‐apoptotic via cyclooxygenase signaling pathway. Based on these findings, aspirin may be used in combination with other chemotherapeutic as a promising treatment of human cervical cancer. Support or Funding Information The study was supported by International Center for Chemical and Biological Sciences, University of Karachi.Fluorescent micrographs of HeLa cells stained with acridine orange/ethidium bromide were used to estimate the apoptotic damage induced by aspirin: (A) Control cells (untreated) exhibiting normal morphology, (B) cells treated with 75 μM (C) cells treated with 100 μM indicating shrinkage of cells and membrane blebbing (D) cells treated with 200 μM show the late early and late phase of apoptosis. The data shown are representative of the combined means from three independent experiments.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .