聚集素，一种在神经母细胞瘤中具有单倍剂量不足特性的肿瘤抑制基因。
Clusterin, a haploinsufficient tumor suppressor gene in neuroblastomas.
作者信息
Chayka Olesya, Corvetta Daisy, Dews Michael, Caccamo Alessandro E, Piotrowska Izabela, Santilli Giorgia, Gibson Sian, Sebire Neil J, Himoudi Nourredine, Hogarty Michael D, Anderson John, Bettuzzi Saverio, Thomas-Tikhonenko Andrei, Sala Arturo
机构信息
Molecular Haematology and Cancer Biology Unit, UCL Institute of Child Health, 30 Guilford St, London WC1N 1EH, UK.
出版信息
J Natl Cancer Inst. 2009 May 6;101(9):663-77. doi: 10.1093/jnci/djp063. Epub 2009 Apr 28.
BACKGROUND
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.
METHODS
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.
RESULTS
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).
CONCLUSIONS
We report, to our knowledge, the first genetic evidence that clusterin is a tumor and metastasis suppressor gene.
背景
在各类人类癌症中，簇集素的表达可能高于或低于正常组织，且簇集素可能促进或抑制细胞凋亡、细胞运动及炎症反应。我们在神经母细胞瘤小鼠模型中研究了簇集素在肿瘤发生发展中的作用。
方法
我们通过实时逆转录-聚合酶链反应评估了MYCN转染的SH-SY5Y和SH-EP细胞中miR-17-92簇微小RNA的表达，并通过转染微小RNA反义寡核苷酸抑制其表达。在携带MYCN转基因和/或簇集素基因杂合或纯合的小鼠（n = 66）中研究肿瘤发生发展情况；这些小鼠来自发生神经母细胞瘤的MYCN转基因小鼠与簇集素基因敲除小鼠的杂交后代。在免疫缺陷小鼠中研究肿瘤生长和转移情况，这些小鼠注射了簇集素表达增强（通过簇集素转染，每组4只小鼠）或降低（通过簇集素短发夹RNA [shRNA]转染，每组8只小鼠）的人神经母细胞瘤细胞。所有统计检验均为双侧检验。
结果
与乱序寡核苷酸相比，用反义寡核苷酸转染抑制SH-SY5Y神经母细胞瘤细胞中MYCN诱导的miR-17-92微小RNA簇表达时，簇集素表达增加。在携带神经母细胞瘤的MYCN转基因小鼠中，发现零个或一个簇集素等位基因的组中患神经母细胞瘤的小鼠在统计学上显著多于有两个簇集素等位基因的组（例如，零等位基因组中有12只患瘤小鼠，而两个等位基因组中有3只，每组n = 22只小鼠；神经母细胞瘤发生的相对风险 = 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过表达簇集素的LA-N-5人神经母细胞瘤细胞比对照细胞少，但在骨髓中发现，转染簇集素shRNA的HTLA230细胞（3.27%，簇集素表达降低）在统计学上显著多于转染对照的细胞（1.53%）（差异 = 1.74%，95% CI = 0.24%至3.24%，P = .026）。
结论
据我们所知，我们首次报告了簇集素是一种肿瘤和转移抑制基因的遗传学证据。
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