Yang Y, Kost-Alimova M, Ingvarsson S, Qianhui Q, Kiss H, Szeles A, Kholodnyuk I, Cuthbert A, Klein G, Imreh S
Microbiology and Tumor Biology Center, Karolinska Institute, Stockholm, Sweden.
Proc Natl Acad Sci U S A. 2001 Jan 30;98(3):1136-41. doi: 10.1073/pnas.98.3.1136.
By passaging microcell hybrids (MCHs) containing human chromosome 3 (chr3) on A9 mouse fibrosarcoma background through severe combined immunodeficient (SCID) mice (elimination test), we have previously defined a 1-Mb-long common eliminated region 1 (CER1) at 3p21.3, a second eliminated region (ER2) at 3p21.1-p14 and a common retained region (CRR) at 3q26-qter. In the present work, chr3 was transferred by microcell fusion into the human nonpapillary renal cell carcinoma line KH39 that contained uniparentally disomic chr3. Four MCHs were generated. Compared with KH39, they developed fewer and smaller tumors, which grew after longer latency periods in SCID mice. The tumors were analyzed in comparison with corresponding MCHs by chr3 arm-specific painting, 19 fluorescent in situ hybridization (FISH) probes, and 27 polymorphic markers. Three MCHs that maintained the intact exogenous chr3 in vitro lost one 3p copy in all 11 tumors. Seven of 11 tumors lost the exogenous 3p, whereas four tumors contained mixed cell populations that lacked either the exogenous or one endogenous KH39 derived 3p. In one MCH the exogenous chr3 showed deletions within CER1 and ER2 already in vitro. It remained essentially unchanged in 8/9 derived tumors. The third, exogenous copy of the 3q26-q27 region (part of CRR) was retained in 16/20 tumors. It can be concluded that the human/human MCH-based elimination test identifies similar eliminated and retained regions on chr3 as the human/murine MCH-based test.
通过在严重联合免疫缺陷（SCID）小鼠中传代培养具有A9小鼠纤维肉瘤背景且包含人类3号染色体（chr3）的微细胞杂种（MCH）（消除试验），我们之前在3p21.3区域定义了一个1兆碱基长的常见消除区域1（CER1），在3p21.1 - p14区域定义了第二个消除区域（ER2），在3q26 - qter区域定义了一个常见保留区域（CRR）。在本研究中，通过微细胞融合将chr3转移到含有单亲二体chr3的人类非乳头状肾细胞癌系KH39中。产生了四个MCH。与KH39相比，它们形成的肿瘤数量更少且体积更小，在SCID小鼠中的潜伏期更长。通过chr3臂特异性染色体涂染、19种荧光原位杂交（FISH）探针和27个多态性标记对肿瘤与相应的MCH进行了分析。在体外维持完整外源chr3的三个MCH在所有11个肿瘤中均丢失了一个3p拷贝。11个肿瘤中有7个丢失了外源3p，而4个肿瘤包含缺乏外源或一个源自KH39的内源3p的混合细胞群体。在一个MCH中，外源chr3在体外CER1和ER2区域内已出现缺失。在8/9个衍生肿瘤中其基本保持不变。3q26 - q27区域（CRR的一部分）的第三个外源拷贝在16/20个肿瘤中得以保留。可以得出结论，基于人/人MCH的消除试验与基于人/鼠MCH的试验在chr3上鉴定出了相似的消除和保留区域。
