Multidrug resistance increment in a human colon carcinoma cell line by colchicine.

The most important mechanism in drug resistance is the multidrug resistance (MDR) phenomenon. It is possible to select MDR cells by in vitro exposure to cytotoxic agents. The resistance is due to the hyperexpression of the P-glycoprotein (P-Gp) that take drugs out from the cells. In this study, a colchicine resistant subline (HCA-2/1cch) was selected from a human colon adenocarcinoma after a short period of drug exposure, as an in vitro model of drug resistance selection. These cells showed cross-resistance to other drugs, which were not present in the medium during selection. The relative resistance was 3.32 for colchicine, 3.15 for vinblastine, 2.62 for vincristine and 5.22 for mitomycin C. P-glycoprotein levels were assayed by flow cytometry. It was found that a significant increase of 2.35 and 1.59 had occurred in the peak and mean channel of fluorescence, respectively, indicating an increment of P-glycoprotein expression in relation to the parental line. Moreover, verapamil (10 microg/ml) produced a partial reversion of multidrug resistance. The sensitisation rates were 7.41 for colchicine, 1.25 for vinblastine, 2.36 for vincristine and 1.17 for mitomycin C. The data obtained suggest that colchicine exposure period (10 weeks) and dose (0.5 microg/ml) assayed were sufficient to produce an increment in multidrug resistance. This resistance could be due to higher level of P-Gp expression.

P-glycoprotein, glutathione and glutathione S-transferase increase in a colon carcinoma cell line by colchicine.

The acquisition of resistance to anticancer agents used in chemotherapy is the main cause of treatment failure in malignant disorders, provoking tumours to become resistant during treatment, although they initially respond to it. The main multidrug resistance (MDR) mechanism in tumour cells is the expression of P-gly-coprotein (P-gly), that acts as an ATP-dependent active efflux pump of chemotherapeutic agents. Furthermore, an increased detoxification of compounds mediated by high levels of glutathione (GSH) and glutathione S-transferase (GST), has been found in resistant cells. We developed a study aiming to evaluate the evolution of the main drug resistance markers in tumour cells: P-gly, GSH and GST, during the acquisition of resistance to colchicine, for the purpose of studying the adaptation process and its contribution to the MDR phenomenon. A human colon adenocarcinoma cell line was exposed to colchicine during 82 days, being P-gly, GSH levels and GST activity evaluated by flow cytometry, spectrofluorimetry and spectrophotometry, during exposure time. P-gly and GSH levels increased gradually during the exposure to colchicine, reaching 2.35 and 3.21 fold each. On day 82, GST activity increased 1.84 fold at the end of the exposure period. Moreover, an increment in drug cross-resistance was obtained that ranges from 2.62 to 5.22 fold for colchicine, vinblastine, vincristine and mitomycin C. The increments obtained in P-gly, GSH and GST could probably contribute to the MDR phenomenon in this human colon adenocarcinoma cell line.

He-Ne laser has no effect on cell cycle phases of human colon adenocarcinoma cells.

The aim of this study is to investigate the effect of He-Ne continuous laser (12.6 mW, 632.8 nm), at low energy densities, on cell cycle synchronization of monolayer growing human colon adenocarcinoma cell line. The doubling time of cell culture was used as optimum time to verify laser effect. The monolayer cultures were exposed to single doses of different energy densities (0.042 J cm-2 to 1.68 J cm-2). The nuclear DNA content has been studied by flow cytometry to obtain the cell percentage in each cell cycle phase. Results show no effect of He-Ne laser irradiation on cell cycle short time synchronization under the previously mentioned conditions and cell type. Higher energy densities and multiple irradiations should be investigated.

[Efficacy of various antihypertensive agents according to age and initial arterial pressure]. / Eficacia de distintos antihipertensivos según la edad y la presión arterial inicial.

210 patients with mild-moderate high blood pressure have been studied. These patients were followed up for 4-6 weeks. Fifty patients were treated with 50 mg hydrochlorothiazide + 5 mg amiloride daily; 40 with 160 mg oxprenolol daily; 22 with 30-40 mg nifedipine daily; 38 with 240 mg verapamil daily and 60 with 50-100 mg captopril daily. We have observed: 1. All treatments have similar antihypertensive effectiveness. 2. Decrement in blood pressure was correlated with initial blood pressure in all treatments, directly. 3. This effectiveness was correlated positively with age in treatment with both diuretics and calcium-antagonists, and an inverse correlation with both beta-blockers and captopril. 4. It had no correlation between age and initial blood pressure in diuretic or beta-blocker treated groups. So diuretics shall be suggested in elderly patients and beta-blockers in younger patients. 5. The greater effectiveness of calcium-antagonists in the elderly should be explained because these patients have a higher initial blood pressure. 6. In this work there is an inverse correlation between age and initial blood pressure in the captopril-treated group; it should explain the greater effectiveness in younger persons. 7. Nothing opposes the use of calcium-antagonists or angiotensin converting enzyme inhibitor in all age groups.

Noradrenaline as a possible mediator of angiotensin II induced fluid transport in rat ileum in vitro.

In isolated segments of ileum excised from bilaterally adrenalectomized and nephrectomized rats, 10(-12) M angiotensin or 10(-3) M noradrenaline added to serosal medium stimulated both fluid transfer and NaCl transport. The alpha adrenergic antagonist phentolamine blocked the stimulation of fluid transfer induced by angiotensin. These results are consistent with the hypothesis that noradrenaline may mediate the increase of intestinal fluid absorption induced by angiotensin in the rat. In segments of isolated ileum from normal rats 10(-12) M angiotensin only stimulated fluid transfer under one of the two following conditions when 10(-3) M imipramine, a noradrenaline uptake blocker, was present in the serosal medium; or when the rats had been previously treated with L-Dopa, a precursor of noradrenaline biosynthesis. These results suggested that the necessity for bilateral adrenalectomy and nephrectomy might be associated to the necessity of increasing the tissue levels of noradrenaline. Direct measurement of noradrenaline tissue content confirmed this.

Stimulatory action of angiotensin II on water and electrolyte transport by the proximal colon of the rat.

In segments of isolated proximal colon from bilaterally adrenalectomized and nephrectomized rats 10(-12) g angiotensin/ml added to the serosal medium stimulated fluid transfer and sodium transport whose increase was accompanied by a decrease in transepithelial potential difference and short circuit current. Transepithelial electrical resistance remained unchanged. Replacement of chloride by sulphate in the bathing medium blocked the response to angiotensin. This suggested that angiotensin stimulated a coupled NaCl transport. Administration of aldosterone also blocked the response to angiotensin, suggesting that a low concentration of endogenous aldosterone is partly responsible for the action of angiotensin. This would explain the necessity for some form of pretreatment such as bilateral adrenalectomy and nephrectomy. Hydraulic conductivity of proximal colon sacs was computed from the variation in the water flow from serosa to mucosa in response to an osmotic shock. A significant (P < 0.01) increase in the hydraulic conductivity of the proximal colon of bilaterally adrenalectomized and nephrectomized rats was found when 10(-12) g angiotensin/ml was present in the serosal bathing medium. The observed decrease in the transepithelial potential difference has been related to an action of angiotensin on the paracellular shunt pathway.