A Novel Mutation of Aryl Hydrocarbon Receptor Interacting Protein Gene Associated with Familial Isolated Pituitary Adenoma Mediates Tumor Invasion and Growth Hormone Hypersecretion
A Novel Mutation of Aryl Hydrocarbon Receptor Interacting Protein Gene Associated with Familial Isolated Pituitary Adenoma Mediates Tumor Invasion and Growth Hormone Hypersecretion
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与家族性孤立性垂体腺瘤 (FIPA) 相关的芳基烃受体相互作用蛋白 (AIP) 基因的新突变介导肿瘤侵袭和生长激素分泌过多
DOI：
10.1016/j.wneu.2018.11.021
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发表时间：
2019-03-01
影响因子：
2
通讯作者：
Zhang, Jianmin
中科院分区：
文献类型：
--
作者：
Cai, Feng;Hong, Yuan;Zhang, Jianmin
背景：在近20%的家族性孤立性垂体腺瘤家族中发现了芳烃受体相互作用蛋白（AIP）基因的种系突变。已证实AIP的某些变异可诱导肿瘤细胞增殖和侵袭性；然而，其机制尚不清楚。方法：在3例中国家族性孤立性垂体腺瘤患者中发现一种新的错义突变（c.512C >t, p.T171I）。在硅和多重连接依赖探针扩增分析预测突变是致病的。采用对照、空白载体、野生型AIP、p. T171I变异体（实验组）、p. Q315*变异体、AIP小干扰RNA等不同载体转染GH3和293FT细胞系，验证该变异体对细胞增殖（cell Counting Kit-8）、侵袭性（Transwell）和生长激素（GH）分泌（酶联免疫吸附法）的影响。同时检测各组Zac1、Sstr2、白细胞介素(IL)-6、Stat3/磷酸化-Stat3的表达（逆转录聚合酶链反应，Western blot）。结果：实验组、p. Q315*变异组、AIP小干扰rna过表达组分别在24、48 h促进细胞增殖（与对照组比较，p < 0.01）。同样，与对照组相比，实验组细胞的侵袭和GH分泌也明显增加（P < 0.01和P < 0.05）。此外，实验组细胞表达较少的Sstr2（对生长抑素类似物的反应性的先决条件）和Zac1（肿瘤抑制基因），但更多的IL-6和磷酸化stat3 （gh分泌相关）。结论：新的AIP突变c. 512C>T （p.T171I）是一种致病变异，通过调节Sstr2、Zac1和IL-6/磷酸化stat3的表达，促进细胞增殖、侵袭性和GH分泌。
BACKGROUND: Germline mutations in the aryl hydrocarbon receptor-interacting protein (AIP) gene were identified in nearly 20% of families with familial isolated pituitary adenoma. Some variants of AIP have been confirmed to induce tumor cell proliferation and invasiveness; however, the mechanism is still unclear.METHODS: A novel missense mutation (c.512C>T, p.T171I) was discovered in 3 patients from a Chinese family with familial isolated pituitary adenoma. In silico and multiplex ligation-dependent probe amplification analysis predicted the mutation to be pathogenic. GH3 and 293FT cell lines were used to verify the variant's effect on cell proliferation (Cell Counting Kit-8), invasiveness (Transwell) and growth hormone (GH) secretion (enzyme-linked immunosorbent assay) by transfection with different vectors: control, blank vector, wild-type AIP, p. T171I variant (experimental group), p. Q315* variant, and AIP small interfering RNA. Furthermore, Zac1, Sstr2, interleukin (IL)-6, and Stat3/phosphorylation-Stat3 expression (reverse transcription polymerase chain reaction, Western blot) in each group was also evaluated.RESULTS: The experimental group, p. Q315* variant group, and AIP small interfering RNA-overexpressing group promoted cell proliferation at 24 and 48 hours, respectively (compared with the control group; P < 0.01 for both). Similarly, the cells in the experimental group manifested more invasion and GH secretion compared with the control group (P < 0.01 and P < 0.05, respectively). Furthermore, the experimental group cells expressed less Sstr2 (a prerequisite for the responsiveness to somatostatin analogues) and Zac1 (tumor suppressor gene), but more IL-6 and phosphorylated-Stat3 (GH-secretion related).CONCLUSIONS: The novel AIP mutation c. 512C>T (p.T171I) is a pathogenic variant that promoted cell proliferation, invasiveness, and GH secretion through regulation of Sstr2, Zac1, and IL-6/phosphorylated-Stat3 expression.