Connexin 37 gene is not mutated in lung carcinomas 3LL and CMT

Our previous studies have shown that intercellular communication mediated by gap junctions is impaired in most tumors as well as in cancer cell lines. However, connexin genes that encode gap junction proteins are only rarely mutated in cancer cells. On the other hand, it was reported that mutated Connexin 37 (Cx37) is the origin of shared tumor-associated antigenic octa-peptides (MUT 1 and MUT 2) of two independently derived lung carcinomas 3LL and CMT 64 of mouse origin. Two Cx37 mutations have been implicated: a Cys-54-Gln substitution in FEQNTAQP (MUT 1) and FEQNTAQA (MUT 2); an additional Pro-59-Ala substitution has been proposed in MUT 2. A Cys-54-Gln mutation in both tumors requires three base changes (TGT-to-CAG) to have occurred twice in independently derived tumors. Another complication stems from the fact that Cys 54, which is located in the extra-cellular domain is conserved in all connexins. Due to the important implications that these findings may have regarding the role of gap junctional communication in lung carcinomas as well as in the origin of tumor-associated antigens, we decided to re-examine these mutations. Thus, we PCR-amplified genomic DNA from 3LL and CMT and sequenced the coding region of Cx37 encompassing codon 54. We then analyzed the PCR products by digestion with the restriction enzyme MaeIII, to discern the presence of the putative mutation. Here we have unambiguously demonstrated that clones Kb39.5 (39.5) and D122 of 3LL, and C6 and E9 of CMT 64, previously employed, have only normal Cx37 sequences, including those of codon 54. Therefore, we concluded that Cx37 is not mutated in 3LL and CMT 64 carcinomas.