肌动蛋白结合蛋白 Anillin 敲低可阻断肝细胞胞质分裂，减少小鼠肝肿瘤发生而不影响再生。
Knockdown of Anillin Actin Binding Protein Blocks Cytokinesis in Hepatocytes and Reduces Liver Tumor Development in Mice Without Affecting Regeneration.
机构信息
Children's Research Institute, Departments of Pediatrics and Internal Medicine, Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, Texas.
Simmons Comprehensive Cancer Center, Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas.
出版信息
Gastroenterology. 2018 Apr;154(5):1421-1434. doi: 10.1053/j.gastro.2017.12.013. Epub 2017 Dec 21.
BACKGROUND & AIMS: Cytokinesis can fail during normal postnatal liver development, leading to polyploid hepatocytes. We investigated whether inhibiting cytokinesis in the liver slows tumor growth without compromising the health of normal hepatocytes. We inhibited cytokinesis in cancer cells by knocking down ANLN, a cytoskeletal scaffolding protein that regulates cytokinesis and might promote tumorigenesis, in mice with liver disease.
METHODS
We analyzed clinical and gene expression data from The Cancer Genome Atlas, Oncomine, PrognoScan, and a hepatocellular carcinoma (HCC) tissue microarray. We knocked down ANLN with small interfering RNAs (siRNAs) in H2.35 liver cells and performed image analyses of cells undergoing cytokinesis. siRNAs were delivered to LAP-MYC mice, which develop hepatoblastoma, using lipid nanoparticles. H2.35 cells with knockdown of ANLN or control cells were injected into FRG mice, which develop chronic liver damage, and tumor growth was monitored. We also developed mice with inducible expression of transgenes encoding small hairpin RNAs (shRNAs) against Anln messenger RNA and studied liver tumorigenesis after administration of diethylnitrosamine and carbon tetrachloride. siRNAs against Anln messenger RNA were conjugated to N-acetylgalactosamine to reduce toxicity and increase hepatocyte tropism; their effects were studied in mouse models of liver cancer and regeneration.
RESULTS
Levels of ANLN messenger RNA were increased in human HCC tissues compared to non-tumor liver tissues. siRNA knockdown of ANLN blocked cytokinesis in H2.35 liver cells. Administration of siRNA against ANLN increased survival times of LAP-MYC mice, compared to mice given a control siRNA. H2.35 liver cells with shRNA knockdown of ANLN formed tumors more slowly in FRG mice than control H2.35 cells. Mice with inducible expression of shRNAs against Anln mRNA developed fewer liver tumors after administration of diethylnitrosamine and carbon tetrachloride than control mice. Knockdown of ANLN did not affect liver regeneration after acute and chronic liver injuries.
CONCLUSIONS
Knockdown of ANLN in liver cells blocks cytokinesis and inhibits development of liver tumors in mice. Agents that inhibit ANLN in the liver might be effective for prevention or treatment of HCC.
背景与目的
细胞分裂在正常出生后肝脏发育过程中可能会失败，导致多倍体肝细胞。我们研究了抑制肝脏中的细胞分裂是否会在不损害正常肝细胞健康的情况下减缓肿瘤生长。我们通过敲低细胞分裂调节蛋白 ANLN 来抑制患有肝病的小鼠的癌细胞中的细胞分裂。
方法
我们分析了癌症基因组图谱、Oncomine、PrognoScan 和肝癌组织微阵列的临床和基因表达数据。我们用小干扰 RNA（siRNA）敲低 H2.35 肝细胞中的 ANLN，并对正在进行细胞分裂的细胞进行图像分析。用脂质纳米粒将 siRNA 递送到 LAP-MYC 小鼠体内，该小鼠会发展成肝癌。用 siRNA 敲低 ANLN 或对照细胞注射 FRG 小鼠，该小鼠会发展成慢性肝损伤，并监测肿瘤生长。我们还开发了可以诱导表达针对 Anln 信使 RNA 的短发夹 RNA（shRNA）的转基因小鼠，并在给予二乙基亚硝胺和四氯化碳后研究肝肿瘤发生情况。将针对 Anln 信使 RNA 的 siRNA 与 N-乙酰半乳糖胺缀合以降低毒性并增加肝细胞趋向性；并在肝癌和再生的小鼠模型中研究了它们的作用。
结果
与非肿瘤性肝组织相比，人肝癌组织中 ANLN 信使 RNA 的水平升高。H2.35 肝细胞中 ANLN 的 siRNA 敲低阻断了细胞分裂。与给予对照 siRNA 的小鼠相比，给予 ANLN 针对 siRNA 的小鼠的存活时间增加。与对照 H2.35 细胞相比，FRG 小鼠中具有 shRNA 敲低 ANLN 的 H2.35 肝细胞形成肿瘤的速度较慢。在给予二乙基亚硝胺和四氯化碳后，可诱导表达针对 Anln mRNA 的 shRNA 的小鼠比对照小鼠形成的肝肿瘤更少。在急性和慢性肝损伤后，ANLN 的敲低对肝再生没有影响。
结论
在肝细胞中敲低 ANLN 可阻断细胞分裂并抑制小鼠肝脏肿瘤的发展。抑制肝脏中的 ANLN 的药物可能对预防或治疗 HCC 有效。
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