Ganglioside GD2 Enhances the Malignant Phenotypes of Melanoma Cells by Cooperating with Integrins.
Ganglioside GD2 Enhances the Malignant Phenotypes of Melanoma Cells by Cooperating with Integrins.
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DOI：
10.3390/ijms23010423
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发表时间：
2021-12-31
影响因子：
5.6
通讯作者：
Furukawa K
中科院分区：
文献类型：
--
作者：
Yesmin F;Bhuiyan RH;Ohmi Y;Yamamoto S;Kaneko K;Ohkawa Y;Zhang P;Hamamura K;Cheung NV;Kotani N;Honke K;Okajima T;Kambe M;Tajima O;Furukawa K;Furukawa K
神经节苷脂被认为可调节细胞膜微区、脂质筏或富含糖脂的微区/筏（GEM/筏）中的细胞信号。特别是，据报道与癌症相关的神经节苷脂会增强癌细胞的恶性特性。事实上，与GD2阴性（GD2 -）细胞相比，GD2阳性（GD2 +）细胞表现出增殖、侵袭和黏附增加。然而，神经节苷脂在GEM/筏中调节细胞信号的精确机制尚不清楚。为了分析神经节苷脂GD2在黑色素瘤细胞恶性特性中的作用，我们使用酶介导的自由基源激活结合质谱法在细胞膜上寻找与GD2相关的分子，并确定整合素β1是一种具有代表性的与GD2相关的分子。然后，我们通过免疫沉淀/免疫印迹证明了GD2和整合素β1的物理关联。免疫细胞染色和邻近连接试验也显示了它们的紧密定位。在细胞黏附过程中，GD2 +细胞显示出多个磷酸化酪氨酸条带，即表皮生长因子受体和黏着斑激酶。整合素β1的敲低表明GD2 +细胞中增加的恶性表型明显被消除。此外，在整合素β1敲低后，GD2 +细胞黏附过程中检测到的磷酸化酪氨酸条带几乎完全消失。最后，通过免疫印迹检查细胞黏附过程中整合素在细胞内的分布，发现在细胞黏附之前和刚刚黏附之后，大量整合素β1定位在GD2 +细胞的GEM/筏组分中，而在GD2 -细胞中大部分定位在非筏组分中。所有这些结果表明，GD2和整合素β1在GEM/筏中协同作用，导致黑色素瘤恶性表型增强。
Gangliosides have been considered to modulate cell signals in the microdomain of the cell membrane, lipid/rafts, or glycolipid-enriched microdomain/rafts (GEM/rafts). In particular, cancer-associated gangliosides were reported to enhance the malignant properties of cancer cells. In fact, GD2-positive (GD2+) cells showed increased proliferation, invasion, and adhesion, compared with GD2-negative (GD2−) cells. However, the precise mechanisms by which gangliosides regulate cell signaling in GEM/rafts are not well understood. In order to analyze the roles of ganglioside GD2 in the malignant properties of melanoma cells, we searched for GD2-associating molecules on the cell membrane using the enzyme-mediated activation of radical sources combined with mass spectrometry, and integrin β1 was identified as a representative GD2-associating molecule. Then, we showed the physical association of GD2 and integrin β1 by immunoprecipitation/immunoblotting. Close localization was also shown by immuno-cytostaining and the proximity ligation assay. During cell adhesion, GD2+ cells showed multiple phospho-tyrosine bands, i.e., the epithelial growth factor receptor and focal adhesion kinase. The knockdown of integrin β1 revealed that the increased malignant phenotypes in GD2+ cells were clearly cancelled. Furthermore, the phosphor-tyrosine bands detected during the adhesion of GD2+ cells almost completely disappeared after the knockdown of integrin β1. Finally, immunoblotting to examine the intracellular distribution of integrins during cell adhesion revealed that large amounts of integrin β1 were localized in GEM/raft fractions in GD2+ cells before and just after cell adhesion, with the majority being localized in the non-raft fractions in GD2− cells. All these results suggest that GD2 and integrin β1 cooperate in GEM/rafts, leading to enhanced malignant phenotypes of melanomas.
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DOI：
10.1073/pnas.90.5.1972
发表时间：
1993-03-01
影响因子：
11.1
作者：
FURUKAWA, K;AKAGI, T;FURUKAWA, K
通讯作者：
FURUKAWA, K
DOI：
10.1073/pnas.0710346105
发表时间：
2008-05-27
影响因子：
11.1
作者：
Kotani, Norihiro;Gu, Jianguo;Honke, Koichi
通讯作者：
Honke, Koichi
影响因子：
7.2
作者：
Kushner, Brian H.;Ostrovnaya, Irina;Cheung, Nai-Kong V.
通讯作者：
Cheung, Nai-Kong V.
DOI：
10.1073/pnas.82.4.1242
发表时间：
1985-01-01
影响因子：
11.1
作者：
HOUGHTON, AN;MINTZER, D;OLD, LJ
通讯作者：
OLD, LJ
影响因子：
3.2
作者：
Hakomori, S
通讯作者：
Hakomori, S