CLONING of TUMOR SUPPRESSOR GENE LOCUS for ENDOMETRIAL CARCINOMA on CHROMOSOME 1

为了鉴定chr. 1上的候选抑制基因的位点，通过微细胞融合将携带间质q臂缺失（del-1 q）或源自chr. 1 q的亚染色体可转移片段（30 A3 STF）的neo标记的人1号染色体转移到人子宫内膜癌细胞系HHUA中。del-1 q和30 A2 STF具有诱导细胞生长停滞和类似衰老细胞的形态学变化的潜力。这些结果表明，对HHUA细胞具有衰老诱导活性的靶基因定位于由D1 S510-D1 S237或D1 S103-D1 S547限定的chr 1 q区域。为了进一步确定候选基因座，我们筛选了61例手术切除的子宫内膜癌样本的缺失重排。最后，我们在1 q41 -42区域发现了一个高发生率的洛丢失，该区域由STS D1 S225和D1 S459定义。在确定了一个保留D1 S225和D1 S459标记的YAC克隆后，我们将其转移到HHUA中，并将其命名为748 H11。YAC克隆还能诱导细胞衰老。利用此YAC DNA或位于YAC内的aBAC DNA，从正常人子宫内膜cDNA文库中分离出几个cDNA克隆。其中，我们发现了一个候选者，其主要序列在许多子宫内膜癌中发生了改变。

To identify the locus of a candidate suppressor gene (s) on chr. 1, A neo-tagged human chromosome 1 that carried an interstitial q arm deletion (del-1q) or a subchromosomal transferrable 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 30A2 STF had the potential to induce the growth arrest and the morphologixal changes analogous to senescent cells. 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, we screened 61 surgically removed endometrial cancer samples for rearrangements of 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 clone, designated 748H11by Research Genetics, which keeps the both D1S225 and D1S459 markers, we transferred it into HHUA. The YAC clone could also induce the cellular senescence. By using this YAC DNA, or aBAC DNA which region is located within the YAC, 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 endometrial cancers.