Pyruvate dehydrogenase kinase 1 and carbonic anhydrase IX targeting in hypoxic tumors.

Pyruvate dehydrogenase kinase 1 (PDHK1) and carbonic anhydrase IX (CAIX) are some of the most hypoxia-inducible proteins associated with tumors, implicated in glucose metabolism and pH regulation, respectively. They both appear to be necessary for model tumor growth, and their high level of expression in human tumors predicts poor patient outcome. Another thing they have in common is that hypoxia not only induces their expression but also their enzymatic activity. This work therefore simultaneously targets these two hypoxia-inducible proteins either pharmacologically or genetically in vitro and in vivo, leading to decreased cancer cell survival and significantly slower model tumor growth. It also suggests that CAIX and PDHK1 are important for cells originating from a colorectal primary tumor, as well as from its metastasis. Moreover, our analysis reveals a unique relationship between these two HIF-1 target genes. In conclusion, the attributes of PDHK1 and CAIX predict them to be promising targets for the design of new, specific inhibitors that could negatively influence tumor cell proliferation and survival, or increase efficacy of standard treatment regimens, and at the same time avoid normal tissue toxicity.

Sequence analysis of the regions flanking terminal repeats of the genome of umava isolate of murine gammaherpesvirus.

The umava isolate of murine gammaherpesvirus (MHV-umava) slightly differs from Murine gammaherpesvirus 68 (MHV-68) and two other isolates of murine gammaherpesvirus (MHV), MHV-76 and MHV-72 in some biological properties. To identify the region(s) in the MHV-umava genome responsible for this phenomenon, we compared the sequences flanking terminal repeats (TRs) of the MHV-umava genome with those of MHV-68, MHV-76 and MHV-72. Restriction and sequence analyses revealed in MHV-umava as compared to MHV-68 approximately 9.3 kbp deletion at the left end of the genome and approximately 1.5 kbp deletion at the right end of the genome. While the approximately 9.3 kbp deletion was similar to that in MHV-76, the approximately 1.5 kbp deletion was unique for MHV-umava.

Lymphocytic choriomeningitis virus mx strain does not induce the expression of tumor-associated carbonic anhydrase IX in persistently infected HeLa cells.

Summary. - Lymphocytic choriomeningitis virus (LCMV) is an arenavirus that readily causes persistent infections, in which it does not interfere with vital functions of the cells, but can affect expression of "luxurious" genes. MX strain of LCMV (MX LCMV) has been identified as an agent transmissible by cell-to-cell contact in the human carcinoma MaTu cells grown in a mixed culture with HeLa cells. When compared to uninfected HeLa, the MaTu-MX-infected HeLa cells, to which the virus was transmitted via co-cultivation with mitomycin C-treated MaTu cells, showed an elevated expression of a protein called MN, suggesting that MN can be induced by MX LCMV. MN protein was later identified as the carbonic anhydrase isoform IX (CA IX), whose expression has been predominantly associated with hypoxic tumors of poor prognosis. Since the proposal that MX LCMV can induce such a cancer-related protein could substantially change our view on the biology of LCMV-host interaction we undertook its verification. Instead of co-cultivation, we used MaTu cell-free extracts to transmit MX LCMV to HeLa cells. These cells were then grown for more than 30 passages, but the level of MN/CA IX did not increase throughout the whole cultivation period as compared to uninfected HeLa cells. Moreover, a treatment of MaTu-MX-infected HeLa cells with ribavirin eliminated the virus, but did not reduce the MN/CA IX expression. Our results clearly showed that MN/CA IX is independent of MX LCMV and that the virus itself does not influence the MN/CA IX level in HeLa cells.