The establishment of chromosome engineering for the gene cloning

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To identify the locus of a candidate suppressor gene(s) on chr. 1 was a model case for the establishment of chromosome engineering for the gene cloning in this study. A neo-tagged human chromosome 1 that carried an interstitial q arm deletion (del-1q) or a subchromosomal transferable fragment (30A3 STF) derived from a chr. 1q was transferred into human endometrial cancer cell line, HHUA via microcell fusion. The del-1q and the 30A3 STF had the potential to induce the growth arrest and the morphological changes analogous to senescent cell. These results indicate that the target gene that has senescence-inducing activity for HHUA cells is localized to the chr. 1q region defined by D1S510-D1S237 or D1S103-D1S547. To further define the candidate locus, the utilization of telomere targeting vector or chromosome pulverization to make finer chromosome fragment was also studied. It seemed useful but more improvements is necessary for the actual usages. Alternatively, we screened 61 surgically removed endometrial cancer samples for rearrangements or deletions. Finally, we found a high incidence of LOH in the 1q41-42 region defined by the STS D1S225 and D1S459. After identifying a YAC (748H11 by Research Genetics) involved in this region, we transferred it into HHUA. The YAC clone also induce the cellular senescence. By using this YAC DNA, several cDNA clones were isolated from the normal human endometrial cDNA library. Among them, we found one candidate which primary sequence was altered in many endrometrial cancers.