Clusterin, a Haploinsufficient Tumor Suppressor Gene in Neuroblastomas
Clusterin, a Haploinsufficient Tumor Suppressor Gene in Neuroblastomas
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DOI：
10.1093/jnci/djp063
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发表时间：
2009-05-06
影响因子：
10.3
通讯作者：
Sala, Arturo
中科院分区：
文献类型：
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作者：
Chayka, Olesya;Corvetta, Daisy;Sala, Arturo
在各种类型的人类癌症中，丛生蛋白的表达可能高于或低于正常组织，并且丛生蛋白可以促进或抑制细胞凋亡、细胞运动和炎症。我们在小鼠神经母细胞瘤模型中研究了clusterin在肿瘤发展中的作用。我们通过实时逆转录-聚合酶链反应在MYCN转染的SH-SY 5 Y和SH-EP细胞中评估了miR-17-92簇中microRNA的表达，并通过转染microRNA反义寡核苷酸抑制其表达。在MYCN转基因和/或丛生蛋白基因杂合或纯合的小鼠（n = 66）中研究肿瘤发展;这些小鼠来自MYCN转基因小鼠（其发展成神经细胞瘤）和丛生蛋白敲除小鼠之间的杂交。在免疫缺陷小鼠中研究肿瘤生长和转移，所述免疫缺陷小鼠注射具有增强（通过clusterin转染，每组4只小鼠）或降低（通过clusterin短发夹RNA [shRNA]转染，每组8只小鼠）clusterin表达的人神经母细胞瘤细胞。所有统计学检验均为双侧检验。与乱序寡核苷酸相比，当转染反义寡核苷酸抑制SH-SY 5 Y神经母细胞瘤细胞中MYCN诱导的miR-17-92 microRNA簇表达时，Escherin表达增加。在统计学上，发现携带MYCN转基因小鼠的神经母细胞瘤在具有零个或一个clusterin等位基因的组中比在具有两个clusterin等位基因的组中显著更多（例如，零等位基因组中12只荷瘤小鼠对双等位基因组中3只荷瘤小鼠，每组n = 22只小鼠;神经母细胞瘤发展的相对风险= 4.85，95%置信区间[CI] = 1.69至14.00; P = .005）。注射后5周，在小鼠肝脏或骨髓中发现比对照细胞更少的clusterin过表达LA-N-5人神经母细胞瘤细胞，但在统计学上显著更多的clusterin shRNA转染的HTLA 230细胞（3.27%，具有降低的clusterin表达）比对照转染细胞（1.53%）（差异= 1.74%，95%CI = 0.24%~ 3.24%，P = 0.026）。
Clusterin expression in various types of human cancers may be higher or lower than in normal tissue, and clusterin may promote or inhibit apoptosis, cell motility, and inflammation. We investigated the role of clusterin in tumor development in mouse models of neuroblastoma.We assessed expression of microRNAs in the miR-17-92 cluster by real-time reverse transcription-polymerase chain reaction in MYCN-transfected SH-SY5Y and SH-EP cells and inhibited expression by transfection with microRNA antisense oligonucleotides. Tumor development was studied in mice (n = 66) that were heterozygous or homozygous for the MYCN transgene and/or for the clusterin gene; these mice were from a cross between MYCN-transgenic mice, which develop neuroblastoma, and clusterin-knockout mice. Tumor growth and metastasis were studied in immunodeficient mice that were injected with human neuroblastoma cells that had enhanced (by clusterin transfection, four mice per group) or reduced (by clusterin short hairpin RNA [shRNA] transfection, eight mice per group) clusterin expression. All statistical tests were two-sided.Clusterin expression increased when expression of MYCN-induced miR-17-92 microRNA cluster in SH-SY5Y neuroblastoma cells was inhibited by transfection with antisense oligonucleotides compared with scrambled oligonucleotides. Statistically significantly more neuroblastoma-bearing MYCN-transgenic mice were found in groups with zero or one clusterin allele than in those with two clusterin alleles (eg, 12 tumor-bearing mice in the zero-allele group vs three in the two-allele group, n = 22 mice per group; relative risk for neuroblastoma development = 4.85, 95% confidence interval [CI] = 1.69 to 14.00; P = .005). Five weeks after injection, fewer clusterin-overexpressing LA-N-5 human neuroblastoma cells than control cells were found in mouse liver or bone marrow, but statistically significantly more clusterin shRNA-transfected HTLA230 cells (3.27%, with decreased clusterin expression) than control-transfected cells (1.53%) were found in the bone marrow (difference = 1.74%, 95% CI = 0.24% to 3.24%, P = .026).We report, to our knowledge, the first genetic evidence that clusterin is a tumor and metastasis suppressor gene.