Singh Praveen, Joon Archana, Kumari Munmun, Singh Tanya, Bal Amanjit, Maan Pratibha, Ghosh Sujata
Department of Experimental Medicine and Biotechnology, PGIMER, Chandigarh, 160012, India.
Department of Histopathology, PGIMER, Chandigarh, 160012, India.
Cell Biochem Biophys. 2022 Dec;80(4):781-793. doi: 10.1007/s12013-022-01091-3. Epub 2022 Sep 9.
Sialylation promotes tumorigenesis by affecting various cancer-related events, including apoptosis inhibition, cell growth, invasion, migration, metastasis, chemo-resistance, and immunomodulation in favor of tumor progression. An altered expression of sialyltransferase enzymes is responsible for synthesizing various tumor-associated sialylated structures. In the present study, our findings have revealed a significant up-regulation of ST3Gal-4 transcript in the two major subtypes of NSCLC cell lines [squamous cell carcinoma cell line (NCI-H520) and adenocarcinoma cell line (A549)]. Thus, the role of the ST3Gal-4 gene was assessed on cancer-associated signal transduction pathways in these cells in view of proliferation, invasion, and migration. ST3Gal-4 was silenced by transfection of both the cell lines with esi-ST3Gal-4-RNA, which RT-PCR and western immunoblotting confirmed. Silencing of ST3Gal-4 resulted in a decreased expression of MAL-I interacting membrane-HSP60, identified earlier as an α2,3-sialylated glycoprotein, thus pointing towards the possible role of ST3Gal-4 in its sialylation. The proliferation, invasion, and migration of both types of NSCLC cells were reduced significantly in the ST3Gal-4 silenced cells. Our findings were substantiated by the down-regulation of β-catenin and E-cadherin, a reduced expression of activated AKT1, ERK1/2, and NF-ƙB in these cells. We propose that ST3Gal-4 may be the disease-associated sialyltransferase involved in α2,3 sialylation of the membrane proteins, including HSP60 of the NSCLC cells. This may lead to the conformational alteration of these proteins, required for the activation of E-cadherin/β-catenin, AKT, and ERK/NF-ƙB mediated signal transduction pathways in these cells, resulting in their proliferation, invasion, and migration.
唾液酸化通过影响各种与癌症相关的事件促进肿瘤发生,这些事件包括抑制细胞凋亡、细胞生长、侵袭、迁移、转移、化疗耐药以及有利于肿瘤进展的免疫调节。唾液酸转移酶的表达改变负责合成各种肿瘤相关的唾液酸化结构。在本研究中,我们的发现揭示了在非小细胞肺癌细胞系的两种主要亚型[鳞状细胞癌细胞系(NCI-H520)和腺癌细胞系(A549)]中ST3Gal-4转录物显著上调。因此,鉴于增殖、侵袭和迁移,对这些细胞中癌症相关信号转导途径上ST3Gal-4基因的作用进行了评估。通过用esi-ST3Gal-4-RNA转染这两种细胞系使ST3Gal-4沉默,RT-PCR和蛋白质免疫印迹证实了这一点。ST3Gal-4的沉默导致MAL-I相互作用膜HSP60的表达降低,MAL-I相互作用膜HSP60先前被鉴定为一种α2,3-唾液酸化糖蛋白,因此表明ST3Gal-4在其唾液酸化中可能发挥的作用。在ST3Gal-4沉默的细胞中,两种类型的非小细胞肺癌细胞的增殖、侵袭和迁移均显著降低。β-连环蛋白和E-钙黏蛋白的下调、这些细胞中活化的AKT1、ERK1/2和NF-ƙB表达的降低证实了我们的发现。我们提出,ST3Gal-4可能是与疾病相关的唾液酸转移酶,参与非小细胞肺癌细胞膜蛋白包括HSP60的α2,3唾液酸化。这可能导致这些蛋白质的构象改变,这是这些细胞中E-钙黏蛋白/β-连环蛋白、AKT和ERK/NF-ƙB介导的信号转导途径激活所必需的,从而导致它们的增殖、侵袭和迁移。