Laboratory of Molecular and Cell Biology, IDI-IRCCS, via Monti di Creta 104, 00167 Rome, Italy.
Genetics and Rare Diseases Research Division, Bambino Gesù Children's Hospital, IRCCS, viale di San Paolo 15, 00146 Rome, Italy.
Int J Mol Sci. 2020 Feb 20;21(4):1426. doi: 10.3390/ijms21041426.
Variably reduced expression of the basement membrane component laminin-332 (α3aβ3γ2) causes junctional epidermolysis bullosa generalized intermediate (JEB-GI), a skin fragility disorder with an increased susceptibility to squamous cell carcinoma (SCC) development in adulthood. Laminin-332 is highly expressed in several types of epithelial tumors and is central to signaling pathways that promote SCC tumorigenesis. However, laminin-332 mutations and expression in individuals affected by JEB-GI and suffering from recurrent SCCs have been poorly characterized. We studied a JEB-GI patient who developed over a hundred primary cutaneous SCCs. Molecular analysis combined with gene expression studies in patient skin and primary keratinocytes revealed that the patient is a functional hemizygous for the p.Cys1171* mutant allele which is transcribed in a stable mRNA encoding for a β3 chain shortened of the last two C-terminal amino acids (Cys1171-Lys1172). The lack of the Cys1171 residue involved in the C-terminal disulphide bond to γ2 chain did not prevent assembly, secretion, and proteolytic processing of the heterotrimeric molecule. Immunohistochemistry of SCC specimens revealed accumulation of mutant laminin-332 at the epithelial-stromal interface of invasive front. We conclude that the C-terminal disulphide bond is a structural element crucial for laminin-332 adhesion function in-vivo. By saving laminin-332 amount, processing, and signaling role the p.Cys1171* mutation may allow intrinsic pro-tumorigenic properties of the protein to be conveyed, thus contributing to invasiveness and recurrence of SCCs in this patient.
层粘连蛋白-332(α3aβ3γ2)的表达可变减少导致交界性大疱性表皮松解症广义中间型(JEB-GI),这是一种皮肤脆弱性疾病,成年后对鳞状细胞癌(SCC)的发展易感性增加。层粘连蛋白-332在几种上皮肿瘤中高度表达,是促进 SCC 肿瘤发生的信号通路的核心。然而,JEB-GI 患者和患有复发性 SCC 的 JEB-GI 患者中发生的层粘连蛋白-332突变和表达特征描述不足。我们研究了一位 JEB-GI 患者,他患有超过一百个原发性皮肤 SCC。分子分析结合患者皮肤和原代角质形成细胞的基因表达研究表明,该患者是 p.Cys1171突变等位基因的功能半合子,该等位基因转录稳定的 mRNA,编码最后两个 C 末端氨基酸(Cys1171-Lys1172)缺失的β3 链。缺乏涉及与 γ2 链的 C 末端二硫键的 Cys1171 残基不阻止三聚体分子的组装、分泌和蛋白水解加工。SCC 标本的免疫组织化学显示突变型层粘连蛋白-332在侵袭前沿的上皮-基质界面处积聚。我们得出结论,C 末端二硫键是层粘连蛋白-332 在体内粘附功能的结构元件。通过保存 p.Cys1171突变的层粘连蛋白-332 量、加工和信号作用,可能允许该蛋白的固有促肿瘤特性得以传递,从而促进该患者 SCC 的侵袭性和复发。