The effects of CD148 Q276P/R326Q polymorphisms in A431D epidermoid cancer cell proliferation and epidermal growth factor receptor signaling.
The effects of CD148 Q276P/R326Q polymorphisms in A431D epidermoid cancer cell proliferation and epidermal growth factor receptor signaling.
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DOI：
10.1002/cnr2.1566
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发表时间：
2022-09
期刊：
影响因子：
1.7
通讯作者：
Takahashi, Takamune
中科院分区：
文献类型：
--
作者：
He, Lilly;Takahashi, Keiko;Pasic, Lejla;Narui, Chikage;Ellinger, Philipp;Grundmann, Manuel;Takahashi, Takamune
CD 148是一种跨膜蛋白酪氨酸磷酸酶，在多种细胞类型中表达。先前的研究表明，CD 148使生长因子受体及其信号分子（包括EGFR和ERK 1/2）去磷酸化，并负调控癌细胞生长。此外，对临床患者的研究表明，高度连锁的CD 148基因多态性Gln 276 Pro（Q276 P）和Arg 326 Gln（R326 Q）与几种癌症的风险增加有关。然而，这些错义突变的生物学效应尚未研究。我们的目的是确定CD 148 Q276 P/R326 Q突变在癌细胞增殖和生长因子信号传导中的生物学效应，重点是EGFR信号传导。将CD 148形式、野生型（WT）或Q276 P/R326 Q逆转录病毒导入缺乏CD 148表达的A431 D表皮样癌细胞中。通过流式细胞术分选表达相当水平的CD 148的稳定细胞。用空逆转录病毒感染的A431 D细胞用作对照。通过免疫染色、细胞增殖试验、酶联免疫吸附试验和蛋白质印迹法评估CD 148定位、细胞增殖率、EGFR信号传导和对血小板反应蛋白-1（TSP 1）（一种CD 148配体）的反应。两种CD 148形式（WT，Q276 P/R326 Q）均分布于细胞表面，所有三种细胞系均表达相同水平的EGFR。与对照细胞相比，表达CD 148形式的A431 D细胞显示出显著较低的细胞增殖率。在这些细胞中，EGF诱导的EGFR和ERK 1/2磷酸化以及细胞增殖也显著降低。此外，TSP 1抑制表达CD 148（WT，Q276 P/R326 Q）的A431 D细胞的细胞增殖，而在对照细胞中没有显示出效果。然而，在CD 148 WT和Q276 P/R326 Q细胞之间未观察到显著差异。我们的数据表明，Q276 P/R326 Q突变对TSP 1-CD 148相互作用以及CD 148的细胞定位和抑制EGFR信号传导和细胞增殖的活性没有重大影响。
CD148 is a transmembrane protein tyrosine phosphatase that is expressed in multiple cell types. Previous studies have shown that CD148 dephosphorylates growth factor receptors and their signaling molecules, including EGFR and ERK1/2, and negatively regulates cancer cell growth. Furthermore, research of clinical patients has shown that highly linked CD148 gene polymorphisms, Gln276Pro (Q276P) and Arg326Gln (R326Q), are associated with an increased risk of several types of cancer. However, the biological effects of these missense mutations have not been studied. We aimed to determine the biological effects of CD148 Q276P/R326Q mutations in cancer cell proliferation and growth factor signaling, with emphasis on EGFR signaling. CD148 forms, wild‐type (WT) or Q276P/R326Q, were retrovirally introduced into A431D epidermoid carcinoma cells that lacks CD148 expression. The stable cells that express comparable levels of CD148 were sorted by flow cytometry. A431D cells infected with empty retrovirus was used as a control. CD148 localization, cell proliferation rate, EGFR signaling, and the response to thrombospondin‐1 (TSP1), a CD148 ligand, were assessed by immunostaining, cell proliferation assay, enzyme‐linked immunosorbent assay, and Western blotting. Both CD148 forms (WT, Q276P/R326Q) were distributed to cell surface and all three cell lines expressed same level of EGFR. Compared to control cells, the A431D cells that express CD148 forms showed significantly lower cell proliferation rates. EGF‐induced EGFR and ERK1/2 phosphorylation as well as cell proliferation were also significantly reduced in these cells. Furthermore, TSP1 inhibited cell proliferation in CD148 (WT, Q276P/R326Q)‐expressing A431D cells, while it showed no effects in control cells. However, significant differences were not observed between CD148 WT and Q276P/R326Q cells. Our data demonstrates that Q276P/R326Q mutations do not have major effects on TSP1‐CD148 interaction as well as on CD148's cellular localization and activity to inhibit EGFR signaling and cell proliferation.