title: Hesperidin inhibits HeLa cell proliferation through apoptosis mediated by endoplasmic reticulum stress pathways and cell cycle arrest.
abstract: BACKGROUND
  Hesperidin (30, 5, 9-dihydroxy-40-methoxy-7-orutinosyl flavanone) is a flavanone that is found mainly in citrus fruits and has been shown to have some anti-neoplastic effects. The aim of the present study was to investigate the effect of hesperidin on apoptosis in human cervical cancer HeLa cells and to identify the mechanism involved.
  
  
  METHODS
  Cells were treated with hesperidin (0, 20, 40, 60, 80, and 100 μM) for 24, 48, or 72 h and relative cell viability was assessed using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay.
  
  
  RESULTS
  Hesperidin inhibited the proliferation of HeLa cells in a concentration- and time-dependent manner. Hesperidin-induced apoptosis in HeLa cells was characterized by increased nuclear condensation and DNA fragmentation. Furthermore, increased levels of GADD153/CHOP and GRP78 indicated hesperidin-induced apoptosis in HeLa cells involved a caspase-dependent pathway, presumably downstream of the endoplasmic reticulum stress pathway. Both of these proteins are hallmarks of endoplasmic reticulum stress. Hesperidin also promoted the formation of reactive oxygen species, mobilization of intracellular Ca(2+), loss of mitochondrial membrane potential (ΔΨm), increased release of cytochrome c and apoptosis-inducing factor from mitochondria, and promoted capase-3 activation. It also arrested HeLa cells in the G0/G1 phase in the cell cycle by downregulating the expression of cyclinD1, cyclinE1, and cyclin-dependent kinase 2 at the protein level. The effect of hesperidin was also verified on the human colon cancer cell HT-29 cells.
  
  
  CONCLUSIONS
  We concluded that hesperidin inhibited HeLa cell proliferation through apoptosis involving endoplasmic reticulum stress pathways and cell cycle arrest.
authors.affiliation: Department of Gynecology, Third Affiliated Hospital of Harbin Medical University, 150 Hapin Road, Harbin, Heilongjiang Province, 150086, China. wangwan_yue@126.com.
authors.forename: Yaoxian
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Cardiopulmonary Function Room, Third Affiliated Hospital of Harbin Medical University, 150 Hapin Road, Harbin, Heilongjiang Province, 150086, China. yuhui197800@yeah.net.
authors.forename: Hui
authors.initials: H
authors.lastname: Yu
authors.affiliation: Department of Gynaecology, Fourth Affiliated Hospital of Harbin Medical University, 37 Yiyuan Street, Harbin, Heilongjiang Province, 150001, China. zhangjin_2000@163.com.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Gynecology, Third Affiliated Hospital of Harbin Medical University, 150 Hapin Road, Harbin, Heilongjiang Province, 150086, China. gaojing150086@yeah.net.
authors.forename: Jing
authors.initials: J
authors.lastname: Gao
authors.affiliation: Department of General Surgery, Heilongjiang Provincial Hospital, 82 Zhongshan Road, Harbin, Heilongjiang Province, 150036, China. gexin628@163.com.
authors.forename: Xin
authors.initials: X
authors.lastname: Ge
authors.affiliation: Department of Gynecology, Third Affiliated Hospital of Harbin Medical University, 150 Hapin Road, Harbin, Heilongjiang Province, 150086, China. gexincom@163.com.
authors.forename: Ge
authors.initials: G
authors.lastname: Lou
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D051033:Apoptosis Inducing Factor / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D019938:Cyclin D1 / Q000378:metabolism; D019927:Cyclin E / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D000091342:Endoplasmic Reticulum Chaperone BiP; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects*; D019073:HT29 Cells; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000378:metabolism; D006569:Hesperidin / Q000494:pharmacology*; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D015513:Oncogene Proteins / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D051742:Transcription Factor CHOP / Q000378:metabolism
chemical_list: D051033:Apoptosis Inducing Factor; C475820:CCNE1 protein, human; D019927:Cyclin E; C497249:DDIT3 protein, human; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; D015513:Oncogene Proteins; D017382:Reactive Oxygen Species; D019938:Cyclin D1; D051742:Transcription Factor CHOP; D045304:Cytochromes c; D006569:Hesperidin; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D053148:Caspase 3; D002118:Calcium