The present paper is focused on zinc(ii) treatment effects on prostatic cell lines PC-3 (tumour) and PNT1A (non-tumour). Oxidative status of cells was monitored by evaluation of expression of metallothionein (MT) isoforms 1A and 2A at the mRNA and protein level, glutathione (oxidised and reduced), and intracellular zinc(ii) after exposition to zinc(ii) treatment at concentrations of 0-150 μM using electrochemical methods, western blotting and fluorescent microscopy. A novel real-time impedance-based growth monitoring system was compared with widely used end-point MTT assay. Impedance-based IC(50) for zinc(ii) is 55.5 and 150.8 μM for PC-3 and PNT1A, respectively. MTT-determined IC(50) are >1.3-fold higher. Impedance-based viability correlates with viable count (r > 0.92; p < 0.03), not with MTT. Two-fold lower intracellular zinc(ii) in the tumour PC-3 cell line was found. After zinc(ii) treatment >2.6-fold increase of intracellular zinc(ii) was observed in non-tumour PNT1A and in tumour PC-3 cells. In PC-3 cells, free and bound zinc(ii) levels were enhanced more markedly as compared to PNT1A. PNT1A produced 4.2-fold less MT compared to PC3. PNT1A cells showed a 4.8-fold increase trend (r = 0.94; p = 0.005); PC-3 did show a significant trend at MT1 and MT2 protein levels (r = 0.93; p = 0.02) with nearly ten-fold increase after 100 μM zinc(ii) treatment. In terms of redox state, PNT1A had a predominance of reduced GSH forms (GSH : GSSG ratio > 1), when exposed to zinc(ii) compared to PC3, where predominance of oxidised forms remains at all concentrations. IC(50) differs significantly when determined by MTT and real-time impedance-based assays due to dependence of impedance on cell morphology and adhesion. When real-time growth monitoring, precise electrochemical methods and fluorescent microscopy are performed together, accurate information for metal fluxes, their buffering by thiol compounds and monitoring of the redox state become a powerful tool for understanding the role of oxidative stress in carcinogenesis.
- MeSH
- buňky - růstové procesy účinky léků MeSH
- fluorescenční barviva chemie MeSH
- fluorescenční mikroskopie metody MeSH
- formazany chemie MeSH
- glutathion metabolismus MeSH
- impedanční spektroskopie MeSH
- lidé MeSH
- lineární modely MeSH
- metalothionein genetika metabolismus MeSH
- nádorové buněčné linie MeSH
- nádory prostaty farmakoterapie metabolismus patologie MeSH
- oxidace-redukce MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- RNA chemie genetika MeSH
- síran zinečnatý farmakologie MeSH
- tetrazoliové soli chemie MeSH
- viabilita buněk fyziologie MeSH
- western blotting MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Photofrin-mediated PDT was applied to malignant (A549 and MCF-7) and normal (HUV-EC-C) cells. The cells were incubated for different lengths of time after PDT. The cell responses to the therapy were examined by changes in SOD activity, phototoxicity, and mode of the cell death. PDT induced dynamic changes in SOD activity. Initially, an increase in SOD activity was observed, and after 6 hours of culture it decreased to the control level. Results obtained from MTT and the comet assay indicate that PDT caused immediate cell death via apoptosis in the A549, MCF-7, and HUV-EC-C cell lines. Our studies confirm that SOD is involved in the response of both cancer and normal cells to PDT.
- MeSH
- dihematoporfyrinether farmakologie MeSH
- formazany MeSH
- fotochemoterapie MeSH
- fotosenzibilizující látky farmakologie MeSH
- kometový test MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory farmakoterapie patologie MeSH
- oxidace-redukce účinky záření MeSH
- senioři MeSH
- superoxiddismutasa metabolismus MeSH
- světlo MeSH
- tetrazoliové soli MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
Aim: A prospective study investigated survival of patients with stage IIIA non-small-cell-lung cancer (NSCLC)treated with a combination of neoadjuvant and adjuvant chemotherapy. Methods: Consecutive chemo-naive patients with potentially operable stage IIIA NSCLC received carboplatin-basedneoadjuvant treatment. Tumor cells harvested during surgery underwent methylthiazolyl tetrazolium blue (MTT)cytotoxic assay. After surgery, adjuvant chemotherapy was selected, where possible, according to MTT results. Results: A total of 65 patients were evaluated (31 received carboplatin/vinorelbine, 34 carboplatin/paclitaxel).The overall response rate was 67.7 % (95% confi dence interval [CI]: 56.3–79.1 %) with downstaging in 52.3 % (95%CI: 40.2–64.5 %) and no signifi cant diff erences between regimens. Median follow-up was 86 months: median overallsurvival (OS) was 32.1 months (95% CI: 7.4–46.5), median time to progression was 25.1 months (95% CI: 15.1–34.9months) and fi ve-year overall survival was 35.7 % (95% CI: 23.7–47.7 %). Forty-seven patients (72.3 %) underwentsurgery and 43 patients received adjuvant chemotherapy. Five-year survival after tumor resection was 49.5 % (95% CI:34.2–64.8 %), median OS was 59.0 months (95% CI: 34.2–83.1) and median disease free survival after surgery was57.3 months (95% CI: 29.5–84.4). With MTT-directed therapy, median OS was 85.1 months (95% CI: 15.4–148.6)and the 5-year survival rate was 57.0 % (95% CI: 34.5–79.5 %); the trend for longer survival failed to reach statisticalsignifi cance. Conclusions: A combination of carboplatin-based neoadjuvant chemotherapy, surgical resection and adjuvantchemotherapy achieved satisfactory survival rates in stage IIIA NSCLC, especially in patients with complete resectionof tumor and those given MTT-directed adjuvant treatment. Our results suggest MTT testing may help optimiseadjuvant chemotherapy.
- MeSH
- adjuvantní chemoterapie metody využití MeSH
- chemorezistence genetika imunologie účinky léků MeSH
- financování organizované MeSH
- formazany diagnostické užití MeSH
- karboplatina terapeutické užití MeSH
- lidé MeSH
- medicína založená na důkazech MeSH
- nemalobuněčný karcinom plic farmakoterapie chirurgie terapie MeSH
- neoadjuvantní terapie metody využití MeSH
- paclitaxel analogy a deriváty terapeutické užití MeSH
- přežití bez známek nemoci MeSH
- prospektivní studie MeSH
- statistika jako téma MeSH
- tetrazoliové soli diagnostické užití MeSH
- vinblastin analogy a deriváty terapeutické užití MeSH
- výsledky a postupy - zhodnocení (zdravotní péče) MeSH
- Check Tag
- lidé MeSH
