Gemcitabine-induced apoptosis in 5637 cell line: An in-vitro model for high-risk superficial bladder cancer

Paola Gazzaniga, Ida Silvestri, Angela Gradilone, Susanna Scarpa, Stefania Morrone, Orietta Gandini, Walter Gianni, Luigi Frati, Anna Maria Aglianò

Research output: Contribution to journalArticle

Abstract

Recent data suggest that new treatment options for superficial bladder cancer are necessary, owing to the high recurrence rate after conventional treatment, especially in T1G3 and Bacillus Calmette-Guerin-refractory patients. Phase I and II studies have demonstrated that gemcitabine may represent a candidate for intravesical therapy in superficial bladder cancer. Despite clinical trials, the in-vitro cytotoxic and proapoptotic effects of gemcitabine have been poorly investigated. In the present study, we investigated how gemcitabine affects apoptosis in bladder cancer cell line 5637, which has the same molecular features of high-risk superficial bladder cancer. Apoptosis was evaluated by DNA fragmentation, flow cytometry and caspase activation. bcl-2, bcl-X, bax, survivin and fas gene expression were also evaluated by reverse-transcriptase polymerase chain reaction. Nuclear factor-kappa B activation was assessed by immunofluorescence. Gemcitabine induced apoptosis in 5637 cells in a time-dependent manner, with activation of caspase-3, -8 and -9. Expression of bcl-2, bax, survivin and bcl-X was not affected by treatment, whereas fas strongly increased after 24 h of treatment. After treatment, we failed to find any nuclear localization of nuclear factor-kappa B. As gemcitabine-induced apoptosis involves fas upregulation, these results may encourage the investigation of intravesical gemcitabine in fas-negative bladder tumors. Furthermore, as nuclear factor-kappa B activation by cisplatin, doxorubicin and adriamycin may result in enhanced proliferation, migration, immortality and inhibition of apoptosis, the observation that gemcitabine does not activate nuclear factor-kappa B may have implications in intravesical therapy of high-risk superficial bladder cancer.

Original languageEnglish
Pages (from-to)179-185
Number of pages7
JournalAnti-Cancer Drugs
Volume18
Issue number2
DOIs
Publication statusPublished - Feb 2007

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Keywords

  • Apoptosis
  • Gemcitabine
  • Nuclear factor-kappa B
  • Superficial bladder cancer

ASJC Scopus subject areas

  • Cancer Research
  • Oncology
  • Pharmacology

Cite this

Gazzaniga, P., Silvestri, I., Gradilone, A., Scarpa, S., Morrone, S., Gandini, O., Gianni, W., Frati, L., & Aglianò, A. M. (2007). Gemcitabine-induced apoptosis in 5637 cell line: An in-vitro model for high-risk superficial bladder cancer. Anti-Cancer Drugs, 18(2), 179-185. https://doi.org/10.1097/CAD.0b013e328010ef47