Induction of chemoresistance in HL-60 cells concomitantly causes a resistance to apoptosis and the synthesis of P-glycoprotein.

The appearance of multidrug-resistant (MDR) proteins or the acquisition of a defective apoptotic programme are major drawbacks in the treatment of cancers since both induce a resistance to classical chemotherapy. However, a link between the two mechanisms has not, as yet, been clearly established. In this study, HL-60 cells cultured in the continual presence of a sub-lethal dose of doxorubicin (dox; HL-60/Dox) were used as a model to study acquired chemoresistance. During the induction of chemoresistance, the appearance of a functional P-glycoprotein (P-gp), in addition to the expression of anti-apoptotic Bcl-2, Bcl-XL and pro-apoptotic Bax proteins was assessed. Parental cells which are sensitive to dox, have no P-gp activity and express Bcl-2 and Bax. After 4 weeks of treatment, a functional P-gp was detected in HL-60/Dox cells. In addition, the synthesis of Bcl-2 appeared to be replaced by Bcl-XL while that of Bax remained unchanged. These cells were also resistant to apoptosis induced by both P-gp and non-P-gp substrates. This inability to induce apoptosis could have resulted from the induction of the expression of the inhibitor of apoptosis protein (XIAP). Our data show that acquired chemoresistance could involve a parallel induction of P-gp and an impairment of the apoptotic pathway.

Production and characterisation of a monoclonal antibody specific for apoptotic bodies derived from several tumour cell lines.

We have recently shown that apoptotic bodies (apobodies) derived from rat colon carcinoma cell lines (PROb) after sodium butyrate (NaB) treatment were able to cure rats with induced peritoneal carcinomatosis ( [BOISTEAU] ). A specific immune response was assumed to be involved since the serum of cured rats contained antibodies specific for apobodies. In the present study, a mAb (clone 6E8) produced by immunisation of rats with apobodies strongly recognized apobodies but had little reactivity with parental tumour cell lines, as demonstrated by enzyme-linked immunosorbent assay (ELISA), immunostaining and flow cytometry. Immunoelectron microscopy showed that 6E8 mAb mainly stained the hyaloplasm or cytosol of apobodies. A protein was detected at 67 kDa by immunoprecipitation of apobodies with mAb, followed by immunoblotting, using serum of rats immunised with apobodies. The 6E8 mAb recognized apobodies derived from several rat or human colon cancer cell lines and a rat glioma cell line, regardless of the apoptosis stimulus used (NaB, staurosporine or UV). Our results clearly show that 6E8 mAb defines an epitope specifically generated during apoptosis, which suggests that the protein recognized may be involved in the molecular cascade of apoptotic cell death.

Regional odontodysplasia: new histopathological data.

Light microscopy, microradiography, SEM and TEM of 4 tooth follicles in a 12-year-old caucasian girl presenting regional odontodysplasia showed widespread globular dentin, calcifications located in the enlarged pulp chambers, hypoplastic and hypomineralized enamel. Hypomineralized strands were sandwiched between two normal enamel layers, which indicates that amelogenesis, interrupted for a while, has once more become established. The enamel surface was covered with calcoglobules. Numerous rounded calcifications were scattered within the dental follicles. Some of these occurred in microfibrils (possibly oxytalan fibers), distinct from collagen fibers. Calcification of the sheath surrounding epithelial rests was a conspicuous feature. The fibroblasts in contact with calcifications developed numerous cytoplasmic extensions, which suggests that they may have assumed a phagocytosis behaviour.

[Dental enamel pearls: histopathological study]. / Les perles d'émail: étude histopathologique.

Enamel pearls are small rounded or egg-shaped nodules which most frequently develop in the furcation area of molars. They may consist of enamel only or enamel containing a small core of tubular dentin. They often sit upon strands of cementum or Tomes granular layer. Histologically, the enamel surface of enamel pearls resembles immature enamel and may be covered with cementum. Small calcoglobules presenting varying degrees of organization are often found related to enamel pearls. Some of them, which have the same radiodensity as the enamel of the pearl and of the affected tooth, may be considered as an enamel-type of calcifications. Others, which present a concentric structure revealed by a polarization cross when viewed between cross polars, may be similar to cementicles. All calcifications located in contact with enamel pearls or at a distance, from them are dystrophic calcifications.