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
- Cell Growth Processes drug effects MeSH
- Fluorescent Dyes chemistry MeSH
- Microscopy, Fluorescence methods MeSH
- Formazans chemistry MeSH
- Glutathione metabolism MeSH
- Dielectric Spectroscopy MeSH
- Humans MeSH
- Linear Models MeSH
- Metallothionein genetics metabolism MeSH
- Cell Line, Tumor MeSH
- Prostatic Neoplasms drug therapy metabolism pathology MeSH
- Oxidation-Reduction MeSH
- Reverse Transcriptase Polymerase Chain Reaction MeSH
- RNA chemistry genetics MeSH
- Zinc Sulfate pharmacology MeSH
- Tetrazolium Salts chemistry MeSH
- Cell Survival physiology MeSH
- Blotting, Western MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study 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
- Dihematoporphyrin Ether pharmacology MeSH
- Formazans MeSH
- Photochemotherapy MeSH
- Photosensitizing Agents pharmacology MeSH
- Comet Assay MeSH
- Middle Aged MeSH
- Humans MeSH
- Cell Line, Tumor MeSH
- Neoplasms drug therapy pathology MeSH
- Oxidation-Reduction radiation effects MeSH
- Aged MeSH
- Superoxide Dismutase metabolism MeSH
- Light MeSH
- Tetrazolium Salts MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female 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
- Chemotherapy, Adjuvant methods utilization MeSH
- Drug Resistance, Neoplasm genetics immunology drug effects MeSH
- Financing, Organized MeSH
- Formazans diagnostic use MeSH
- Carboplatin therapeutic use MeSH
- Humans MeSH
- Evidence-Based Medicine MeSH
- Carcinoma, Non-Small-Cell Lung drug therapy surgery therapy MeSH
- Neoadjuvant Therapy methods utilization MeSH
- Paclitaxel analogs & derivatives therapeutic use MeSH
- Disease-Free Survival MeSH
- Prospective Studies MeSH
- Statistics as Topic MeSH
- Tetrazolium Salts diagnostic use MeSH
- Vinblastine analogs & derivatives therapeutic use MeSH
- Outcome and Process Assessment, Health Care MeSH
- Check Tag
- Humans MeSH
- MeSH
- Precursor Cell Lymphoblastic Leukemia-Lymphoma drug therapy MeSH
- Child MeSH
- Formazans MeSH
- Drug Resistance MeSH
- Humans MeSH
- Tumor Cells, Cultured drug effects MeSH
- Antineoplastic Agents pharmacology MeSH
- Recurrence drug therapy MeSH
- In Vitro Techniques MeSH
- Tetrazolium Salts MeSH
- Check Tag
- Child MeSH
- Humans MeSH
