Comparison of P-glycoprotein expression with in vitro drug sensitivity in fresh blast cells from acute myeloid leukaemia patients.

Anthracyclines and etoposide have been implicated in the multi-drug resistance phenotype. The mdr 1 gene encodes for the transmembrane protein P-glycoprotein. P-glycoprotein expression was measured in the fresh blast cells from 19 patients with acute myeloid leukemia using three monoclonal antibodies, C219, JSB-1 and MRK 16, and immunocytochemistry with the enzyme alkaline phosphatase as marker. Drug resistance can be identified in vitro using the predictive chemosensitivity test, the MTT (3-4,5-dimethylthiazol-2,5-diphenyl tetrazolium bromide) assay. In order to assess cell viability after drug exposure, this technique utilises the ability of cellular dehydrogenase enzymes to reduce the tetrazolium salt MTT to formazan. In vitro resistance to multi-drug resistance related cytotoxic agents was identified in the blast cells from these patients. This study showed no correlation between the results of the MTT assay and P-glycoprotein expression in this disease, suggesting either that more sensitive techniques are required to measure P-glycoprotein expression or that other drug resistance mechanisms may be involved.

Metabolism of the anti-depressant lortalamine.

Humans excreted an oral dose of 5 mg of the anti-depressant lortalamine (radiolabelled with carbon-14) mainly in the urine (98% during 5 days). Plasma 14C concentrations were highest (about 44 ng equiv./ml) between 1.5-3 h when the corresponding concentrations of unchanged drug were about 17 ng/ml. Unchanged drug concentrations appeared to decline with a half-life of about 5 h. Concentrations of 14C then declined rapidly and were below the limits of detection (12 ng equiv./ml) at 24-36 h. Measurement of whole-blood 14C concentrations showed that there was some uptake into the blood cells (about 65% of the peak plasma 14C level). Radioactivity in the urine was mainly associated with unchanged drug and three metabolites. The major metabolite (about 50% urinary 14C) was identified by mass spectrometry as the N-demethylated compound and another metabolite as a keto derivative of lortalamine where oxidation had occurred in the piperidine ring.

Acquisition of a limited lifespan by differentiating cells derived from PC13 embryonal carcinoma cells.

Retinoic acid (RA) has previously been shown to induce the differentiation of mouse embryonal carcinoma (EC) cells to endoderm-like cells that have a slower rate of proliferation and are nontumorigenic. These cells also acquire the ability to respond to a range of exogenous growth factors. We have analysed the change in growth phenotype for PC13 EC cells using video recordings and autoradiography. We have shown that the endoderm-like cells have a longer cell cycle time than their undifferentiated counterparts (five cell divisions after exposure to RA the differentiated cells had a median cell cycle time of 1800 min compared to 800 min for control cells). The endoderm-like cells also have a progressively decreasing probability of dividing again and this indicates that the differentiation process is accompanied by the acquisition of a limited life-span. The characteristics of mortal cells are well documented, and the endoderm-like cells demonstrate the properties of such cells. In addition, we have confirmed the observation that epidermal growth factor (EGF) can stimulate the proliferation of the endoderm-like cells and have shown, using autoradiography, that 92% of these cells express EGF receptors. Using video recordings, we have demonstrated that the effect of EGF is to shorten the cell cycle of the differentiating cells. We have also shown that EGF can enhance the survival of the endoderm-like cells and thereby prolong their life-span. It is known that EGF and other growth factors can prolong the life-span of mortal cells derived from normal tissues, but we have demonstrated that EGF can have this effect on the differentiated derivatives of a tumour cell.