Abstract：
Shwachman-Diamond syndrome (SDS) is an autosomal recessive disorder characterized by exocrine pancreatic insufficiency and bone marrow failure. The depletion of SBDS protein by RNA interference has been shown to cause inhibition of cell proliferation in several cell lines. However, the precise mechanism by which the loss of SBDS leads to inhibition of cell growth remains unknown. To evaluate the impaired growth of SBDS-knockdown cells, we analyzed Epstein-Barr virus-transformed lymphoblast cells (LCLs) derived from two patients with SDS (c. 183_184TA > CT and c. 258 + 2 T > C). After 3 days of culture, the growth of LCL-SDS cell lines was considerably less than that of control donor cells. By annealing control primer-based GeneFishing PCR screening, we found that galectin-1 (Gal-1) mRNA expression was elevated in LCL-SDS cells. Western blot analysis showed that the level of Gal-1 protein expression was also increased in LCL-SDS cells as well as in SBDS-knockdown 32Dcl3 murine myeloid cells. We confirmed that recombinant Gal-1 inhibited the proliferation of both LCL-control and LCL-SDS cells and induced apoptosis (as determined by annexin V-positive staining). These results suggest that the overexpression of Gal-1 contributes to abnormal cell growth in SBDS-deficient cells.
摘要：
Shwachman-Diamond综合征（SDS）是一种常染色体隐性遗传疾病，其特征是胰腺外分泌功能不全和骨髓衰竭。通过RNA干扰的SBDS蛋白的消耗已被证明在几种细胞系中引起细胞增殖的抑制。然而,SBDS丢失导致细胞生长抑制的确切机制仍然未知。为了评估SBDS敲低细胞的生长受损，我们分析了来自两名SDS患者的EB病毒转化的淋巴母细胞（LCLs）（c.183_184TA>CT和c。258+2T>C)。培养3天后,LCL-SDS细胞系的生长明显低于对照供体细胞。通过基于退火对照引物的GeneFishingPCR筛选,我们发现半乳糖凝集素-1（Gal-1）mRNA在LCL-SDS细胞中表达升高。Western印迹分析显示,Gal-1蛋白表达水平在LCL-SDS细胞以及SBDS敲低32Dcl3鼠骨髓细胞中也增加。我们证实了重组Gal-1抑制了LCL对照和LCL-SDS细胞的增殖并诱导了细胞凋亡（通过膜联蛋白V阳性染色确定）。这些结果表明Gal-1的过表达有助于SBDS缺陷细胞中的异常细胞生长。
