|Cancer||Cell death||Cell cycle||Cytoskeleton||Exo/endocytosis||Differentiation||Division||Organelles||Signalling||Stem cells||Trafficking|
Multiple factors influencing the release of hTERT mRNA from pancreatic cancer cell lines in in vitro culture
Gensaku Okada, Hiroyuki Watanabe, Koushiro Ohtsubo1, Hisatsugu Mouri, Yasushi Yamaguchi, Yoshiharu Motoo and Norio Sawabu
Division of Medical Oncology, Kanazawa University Cancer Research Institute, 131, Takaramachi, Kanazawa, Ishikawa 9200934, Japan
Since telomerase expression is highly prevalent in human cancers, the quantitation of serum/plasma hTERT (human telomerase reverse transcriptase) mRNA levels may be useful for early detection of PCa (pancreatic cancer). To analyse the correspondence between exhTERT (extracellular hTERT) mRNA levels and hTERT expression, we designed a cell culture system to investigate factors modulating the extracellular levels of hTERT mRNA in media conditioned by eight PCa cell lines. We found that the level of exhTERT mRNA was dependent on cell growth rate. MIAPaCa-2, PANC-1, KLM-1 and PK-9 cells expressed high levels of exhTERT mRNA, independent of cell density, whereas proliferating PK-59, BxPC-3 and PK-45H cells released low levels of exhTERT mRNA. The augmented release of mRNA by spontaneous dead MIAPaCa-2 cells was further increased at postconfluence. In Capan-1 cells, low correspondence of marker was also due to RNase secretion. Upon reaching confluence, some PCa cell lines showed down-regulation of hTERT expression. Following cell–cell adhesion, as shown by E-cadherin engagement, PK-59 cells showed levels of extracellular message below the limits of detection, a loss not due to an increase in message degradation. These results suggest that the levels of exhTERT mRNA in the medium of PCa cell lines are altered not only in response to cell growth rate and cell destruction, but are responsive to extracellular cues such as RNases and cell density. A cell-free assay for exhTERT mRNA may therefore not be useful for early detection of PCa.
Key words: cell–cell adhesion, conditioned medium, extracellular hTERT mRNA, pancreatic cancer, real-time RT-PCR, tumour marker
Abbreviations: DMEM, Dulbecco's modified Eagle's medium, exhTERT, extracellular hTERT, FBS, fetal bovine serum, H, high cell density, hTERT, human telomerase reverse transcriptase, IgG, immunoglobulin G, L, low cell density, mAb, monoclonal antibody, PCa, pancreatic cancer, RT-PCR, reverse transcription-PCR, SF, serum-free
1To whom correspondence should be addressed (email email@example.com).
Received 26 December 2009/14 May 2010; accepted 16 November 2010
Published as Cell Biology International Immediate Publication 16 November 2010, doi:10.1042/CBI20090471
© The Author(s) Journal compilation © 2012 International Federation for Cell Biology