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Increased adenoviral transduction efficacy in human laryngeal carcinoma cells resistant to cisplatin is associated with increased expression of integrin α v β 3 and coxsackie adenovirus receptor
Author(s) -
AmbriovićRistov Andreja,
Gabrilovac Jelka,
ČimboraZovko Tamara,
Osmak Maja
Publication year - 2004
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.20176
Subject(s) - transduction (biophysics) , biology , signal transduction , flow cytometry , microbiology and biotechnology , cisplatin , downregulation and upregulation , adenoviridae , viral vector , cell culture , cancer research , genetic enhancement , gene , genetics , biochemistry , recombinant dna , chemotherapy
In our study, we investigated molecular mechanisms of increased adenoviral transduction efficacy in cisplatin‐resistant human laryngeal carcinoma cells CA3 ST as compared to parental cells HEp2. Using reverse transcription‐PCR, the genes potentially implicated in adenoviral entry were screened. In cisplatin‐resistant cells, only upregulation of α v β 3 integrin was detected, which was additionally confirmed by flow cytometry. Moderately increased expression of CAR was determined in cisplatin‐resistant CA3 ST cells using flow cytometry and measurement of wild‐type adenovirus Ad5CMVβgal attachment. In order to test the implication of α v β 3 integrin in transduction efficacy, 6 HEp2‐derived α v β 3 ‐expressing clones with graded expression of α v β 3 were isolated. To a certain degree of density, expression of α v β 3 positively correlated with Ad5CMVβgal transduction efficacy ( i.e ., increased viral transduction), suggesting a role of α v β 3 in transduction efficacy. However, HEp2 clones with the highest α v β 3 expression were negatively correlated with transduction efficacy ( i.e ., decreased viral transduction). This was shown to be associated with downregulation of α v β 5 integrin, also involved in viral transduction, in clones with the highest α v β 3 expression. The implication of CAR in increased adenoviral transduction efficacy in cisplatin resistant CA3 ST cells was further assessed by transduction experiments using adenoviral mutant Ad5FbΔ639 whose entry is only to a very small extent dependent on the presence of CAR. Indeed, Ad5FbΔ639 infected 2.5‐fold more, in comparison to wild‐type adenovirus, which infected 5‐fold more efficiently resistant CA3 ST cells than parental HEp2 cells, indicating that increased expression of CAR contributes to increased efficacy of adenoviral transduction. Thus, the data presented provide evidence that both α v β 3 integrin and CAR are involved in increased adenoviral transduction efficacy in cisplatin resistant CA3 ST cells. These findings may have significant implications in human gene therapy using adenoviruses, especially in patients after unsuccessful cisplatin treatment. © 2004 Wiley‐Liss, Inc.