estrone-sulfate and cholesteryl-sulfate

In this work, we studied indolium and benzothiazolium pentamethine salts 1-3 as novel type of receptors for the recognition of sulphated signalling molecules (sulphated steroids: oestrone, pregnenolone and cholesterol sulphate). A recognition study was performed in an aqueous medium (1mM phosphate buffer (H2O:MeOH; 99:1 (v/v))) at pH 7.34. The tested salts displayed a high affinity for these sulphated analytes, mainly for cholesterol sulphate. However, no interaction between the salts and control, non-sulphated sterol analytes (cholesterol and bile acid) was observed. The highest affinity for the sulphated steroids was observed for benzothiazole salt 1. This salt also displayed different spectral behaviour from that observed for carbocyanine salts 2 and 3. In this presence of cholesterol sulphate, benzothiazole salt 1 displayed significant spectral changes depending on the medium used: a blue shift in the aqueous medium and a red shift in the methanolic one (H2O:MeOH; 2:1 (v/v)). Subsequently preliminary in vivo study showed that, salt 1 significantly inhibits a growth of breast carcinoma on Nu/nu mice model.

Topics: Animals; Antineoplastic Agents; Benzothiazoles; Breast Neoplasms; Carbocyanines; Cholesterol Esters; Estrone; Female; Heterocyclic Compounds; Mice, Nude; Pregnenolone; Xenograft Model Antitumor Assays

Although the exact function of cholesteryl sulfate (CS) is unknown, it is present in low concentration in lipoproteins, in red blood cells and spermatozoa. In the present study, we investigated whether CS is present in blood platelets and its possible biological involvement in platelet function. Extensively washed platelets were prepared from rat and human blood. After lipid extraction and thin layer chromatography (TLC) on silica gel, a compound with the same mobility as authentic CS was isolated and identified by two different methods: (1) without hydrolysis, negative ion fast atom bombardment combined with tandem mass spectrometry (MS/MS); (2) after acidic hydrolysis, identification of cholesterol (Chol) by TLC and gas chromatography-MS. CS concentrations measured using beta-sitosteryl sulfate as internal standard in normal rat or human platelets were in the range of 164-512 pmol/10(9) platelets. This represented less than 1% of cell Chol. Biological effects of CS on platelet function were studied in vitro. CS incubated with rat platelets either as methanol solution or as albumin-bound complex potentiated the ADP- or thrombin-induced aggregation and serotonin secretion. The results of platelet sterol analysis indicated that CS was incorporated into platelet membrane and did not significantly change the platelet cholesterol composition. The potentiating effect of CS on platelet-induced aggregation and secretion was not obtained with cholesterol, cholesteryl acetate or estrone. In contrast, an inhibitory effect of estrone sulfate was observed. These results indicate that both the sulfate group and the cholesterol moiety are involved in the pro-aggregant property of CS. In addition, platelet mediators seem to be implicated in the mechanism since the thrombin-induced production of thromboxane B2, the stable end-product of arachidonic acid metabolism, was also enhanced in the presence of CS. These results suggest a new role for CS which may be involved in the modulation of platelet function.

Topics: Animals; Blood Platelets; Cholesterol; Cholesterol Esters; Chromatography, Thin Layer; Estrone; Humans; Male; Platelet Aggregation; Rats; Serotonin; Spectrometry, Mass, Fast Atom Bombardment; Thrombin; Thromboxane B2