Laboratory of Physiological Chemistry, Hiroshima International University, 5-1-1 Hirokoshingai, Kure-Shi, Hiroshima, 737-0112, Japan.
Department of Child Health and Development, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, BUnkyo-ku, Tokyo, 113-8519, Japan.
Int J Hematol. 2024 Apr;119(4):383-391. doi: 10.1007/s12185-024-03709-z. Epub 2024 Jan 19.
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.
Shwachman-Diamond 综合征(SDS)是一种常染色体隐性疾病,其特征为外分泌胰腺功能不全和骨髓衰竭。通过 RNA 干扰耗竭 SBDS 蛋白已被证明可抑制几种细胞系的细胞增殖。然而,SBDS 缺失导致细胞生长抑制的确切机制尚不清楚。为了评估 SBDS 敲低细胞的生长受损情况,我们分析了来自两名 SDS 患者(c.183_184TA>CT 和 c.258+2T>C)的 Epstein-Barr 病毒转化的淋巴母细胞(LCL)。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 GeneFishing PCR 筛选,我们发现 LCL-SDS 细胞中半乳糖凝集素-1(Gal-1)mRNA 的表达升高。Western blot 分析显示,Gal-1 蛋白表达水平在 LCL-SDS 细胞以及 SBDS 敲低 32Dcl3 鼠髓样细胞中也升高。我们证实重组 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 annexin V 阳性染色确定)。这些结果表明 Gal-1 的过表达有助于 SBDS 缺陷细胞的异常细胞生长。