Division of Clinical Chemistry, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand.
Biomedical Engineering Program, Faculty of Engineering, Chiang Mai University, Chiang Mai, 50200, Thailand.
Sci Rep. 2019 Jul 17;9(1):10394. doi: 10.1038/s41598-019-46701-y.
Glucosidase II (GluII) plays a major role in regulating post-translation modification of N-linked glycoproteins. We have previously reported that the expression of glucosidase II beta subunit (GluIIβ) was significantly increased in lung tumor tissues and its suppression triggers autophagy and/or apoptosis. Here, we investigated the role of GluIIβ in cell growth, metastatic potential, and receptor tyrosine kinases (RTKs) signaling activity in lung carcinoma cell lines. CRISPR-CAS9 technology was used to knockout the GluIIβ encoding gene (PRKSH) in lung carcinoma cells. GluIIβ knockout cells exhibited drastically slower growth rates in comparison to non-target transfected cells, particularly with lower concentrations of fetal bovine serum, indicating impairment of their ability to survive under nutritional deprivation. Cell migration and anchorage-independent growth, the fundamental components of cancer cell metastasis, were significantly decreased in GluIIβ knockout cells. Knockout of GluIIβ increased the sensitivity of lung cancer cells to cisplatin but reduced their sensitivity to gefitinib. Interestingly, knocking out of GluIIβ lowered overall RTK signaling activities to less than half of those in non-target transfected cells, which could represent a novel strategy for blocking multiple RTKs in tumor cells in an effort to improve lung cancer treatment.
葡糖苷酶 II（GluII）在调节 N-连接糖蛋白的翻译后修饰中起着主要作用。我们之前曾报道过，葡糖苷酶 II β亚基（GluIIβ）在肺癌组织中的表达显著增加，其抑制会触发自噬和/或细胞凋亡。在这里，我们研究了 GluIIβ 在肺癌细胞系中的细胞生长、转移潜能和受体酪氨酸激酶（RTKs）信号活性中的作用。使用 CRISPR-CAS9 技术敲除了肺癌细胞中编码 GluIIβ 的基因（PRKSH）。与非靶向转染细胞相比，GluIIβ 敲除细胞的生长速度明显较慢，特别是在胎牛血清浓度较低的情况下，表明它们在营养剥夺下的生存能力受损。细胞迁移和无锚定生长是癌细胞转移的基本组成部分，在 GluIIβ 敲除细胞中显著降低。敲除 GluIIβ 增加了肺癌细胞对顺铂的敏感性，但降低了它们对吉非替尼的敏感性。有趣的是，敲除 GluIIβ 会将总体 RTK 信号活性降低至非靶向转染细胞的一半以下，这可能代表了一种在肿瘤细胞中阻断多种 RTKs 的新策略，以努力改善肺癌治疗效果。
