title: Effect of aldosterone on the amplification of oncolytic vaccinia virus in human cancer lines.
abstract: OBJECTIVE
  JX-594 is an oncolytic virus derived from the Wyeth vaccinia strain that causes replication-dependent cytolysis and antitumor immunity. Starting with a cross-examination of clinical-trial samples from advanced hepatocellular carcinoma patients having high levels of aldosterone and virus amplification in JX-594 treatment, we investigated the association between virus amplification and aldosterone in human cancer cell lines.
  
  
  METHODS
  Cell proliferation was determined by a cell-counting-kit-based colorimetric assay, and vaccinia virus quantitation was performed by quantitative polymerase chain reaction (qPCR) and a viral plaque assay. Also, the intracellular pH was measured using a pH-sensitive dye.
  
  
  RESULTS
  Simultaneous treatment with JX-594 and aldosterone significantly increased viral replication in A2780, PC-3, and HepG2 cell lines, but not in U2OS cell lines. Furthermore, the aldosterone treatment time altered the JX-594 replication according to the cell line. The JX-594 replication peaked after 48 and 24 hours of treatment in PC-3 and HepG2 cells, respectively. qPCR showed that JX-594 entry across the plasma membrane was increased, however, the changes are not significant by the treatment. This was inhibited by treatment with spironolactone (an aldosterone-receptor inhibitor). JX-594 entry was significantly decreased by treatment with EIPA [5-(N-ethyl-N-isopropyl)amiloride; a Na(+)/H(+)-exchange inhibitor], but aldosterone significantly restored JX-594 entry even in the presence of EIPA. Intracellular alkalization was observed after aldosterone treatment but was acidified by EIPA treatment.
  
  
  CONCLUSIONS
  Aldosterone stimulates JX-594 amplification via increased virus entry by affecting the H(+) gradient.
authors.affiliation: Department of Pharmacology, Pusan National University School of Korean Medicine, Busan, Korea.
authors.forename: Hyun Ju
authors.initials: HJ
authors.lastname: Lee
authors.forename: Jasung
authors.initials: J
authors.lastname: Rho
authors.forename: Shao Ran
authors.initials: SR
authors.lastname: Gui
authors.forename: Mi Kyung
authors.initials: MK
authors.lastname: Kim
authors.forename: Yu Kyoung
authors.initials: YK
authors.lastname: Lee
authors.forename: Yeon Sook
authors.initials: YS
authors.lastname: Lee
authors.forename: Jeong Eun
authors.initials: JE
authors.lastname: Kim
authors.forename: Euna
authors.initials: E
authors.lastname: Cho
authors.forename: Mong
authors.initials: M
authors.lastname: Cho
authors.forename: Tae Ho
authors.initials: TH
authors.lastname: Hwang
mesh_terms: D000450:Aldosterone / Q000494:pharmacology*; D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000821:virology; D045744:Cell Line, Tumor; D006801:Humans; D006854:Hydrocortisone / Q000097:blood; D006863:Hydrogen-Ion Concentration; D008113:Liver Neoplasms / Q000097:blood / Q000821:virology; D000451:Mineralocorticoid Receptor Antagonists / Q000494:pharmacology; D018696:Neuroprotective Agents / Q000494:pharmacology; D050130:Oncolytic Virotherapy; D011817:Rabbits; D013148:Spironolactone / Q000494:pharmacology; D014616:Vaccinia virus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000451:Mineralocorticoid Receptor Antagonists; D018696:Neuroprotective Agents; D013148:Spironolactone; D000450:Aldosterone; D000584:Amiloride; C039614:ethylisopropylamiloride; D006854:Hydrocortisone
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