Human multidrug resistant KB cells overexpress protein kinase C: involvement in drug resistance

Among the many phenotypic characteristics of multidrug resistance (MDR), the presence of P-glycoprotein is nearly always observed, and it appears that the plasma membrane of the multidrug resistant cell is integrally involved in controlling drug resistance. Another membrane-associated protein kinase, protein kinase C (PKC), has been shown to regulate the flow of information to the cell interior and to control the efflux of a number of different compounds. We therefore initiated a study of PKC and MDR. We found that multidrug resistant sublines from both mouse sarcoma 180 and human KB lines exhibited 80-90% increases in basal PKC activity. The mechanism of the increase appears to be quite different in the two cell lines. The human KB cells overexpress the alpha isozyme of PKC, commensurate with the increase in alpha-PKC protein, whereas the mouse cells do not overexpress alpha-mRNA but increase alpha-PKC protein. Furthermore, it appears that PKC activity plays a functional role in drug resistance, since inhibition of endogenous PKC activity by staurosporine resulted in decreased resistance to Adriamycin. We also found that phosphorylation of MDR cell membrane vesicles by purified PKC, followed by immunoprecipitation of P-glycoprotein with monoclonal antibody C219, resulted in a level of phosphorylation of P-glycoprotein that was greater than the endogenous phosphorylation level. The data presented indicate that MDR cells of diverse species exhibited enhanced PKC activity but that the mechanisms were different. The increased kinase activity may have biological relevance to MDR since PKC appears to be coupled to P-glycoprotein function.     

Pancreatic transplant imaging

Forty-four clinical episodes of suspected (pancreas) transplant rejection in 17 pancreatic transplantation patients were reviewed retrospectively. The clinical impression of acute graft rejection, chronic rejection, or nonrejection in each episode was correlated with the results of 19 nuclear medicine, 12 ultrasound (US), and 44 magnetic resonance (MR) imaging studies. US was found to be a moderately sensitive (82%) method of detecting graft rejection. US also was effective in identifying intra- and peripancreatic fluid accumulations. Nuclear medicine imaging was also a sensitive technique (86%) and the only modality that provided physiologic information regarding graft perfusion. MR imaging allowed correct prediction of the presence or absence of graft rejection in 39 of 44 cases (sensitivity, 100%; specificity, 76%) and was an effective means of detecting pathologic fluid collections. Nuclear medicine, US, and MR imaging are all believed to be sensitive methods of detecting graft rejection and are complementary adjuncts to the clinical evaluation of pancreatic transplants.