Oncogenic transformation of NIH/3T3 cells by the overexpression of L-type amino acid transporter 1, a promising anti-cancer target.
Oncogenic transformation of NIH/3T3 cells by the overexpression of L-type amino acid transporter 1, a promising anti-cancer target.
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DOI：
10.18632/oncotarget.27981
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发表时间：
2021-06-22
期刊：
影响因子：
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通讯作者：
Sugiura R
中科院分区：
文献类型：
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作者：
Hayashi N;Yamasaki A;Ueda S;Okazaki S;Ohno Y;Tanaka T;Endo Y;Tomioka Y;Masuko K;Masuko T;Sugiura R
L型氨基酸转运蛋白1（LAT 1）/SLC 7A 5是第一个鉴定的与CD 98重链（CD 98 hc/SLC 3A 2）二硫键连接的CD 98轻链。LAT 1转运大的中性氨基酸，包括激活mTOR的亮氨酸，并在人类癌症中高度表达。我们研究了通过逆转录病毒感染将人LAT 1导入NIH/3 T3细胞的致癌性。建立了稳定表达人天然（164 C）或突变（164 S）LAT 1（分别为naLAT 1/3 T3或muLAT 1/3 T3）的NIH/3 T3细胞系。我们证实了内源性小鼠CD 98 hc在naLAT 1/3 T3中与外源性人LAT 1形成二硫键，但在muLAT 1/3 T3中不形成二硫键。内源性小鼠CD 98 hc mRNA在naNIH/3 T3和muLAT 1/3 T3中均增加，并且在两种细胞系中检测到相似量的外源性人LAT 1蛋白。此外，评价了naLAT 1/3 T3和muLAT 1/3 T3细胞系的细胞生长相关表型（ERK磷酸化、细胞周期进展）和细胞恶性相关表型（非贴壁依赖性细胞生长、裸鼠肿瘤形成）。naLAT 1/3 T3具有比NIH/3 T3和muLAT 1/3 T3更强的生长和恶性相关表型，表明天然LAT 1通过其与CD 98 hc的相互作用的致癌性。抗LAT 1单克隆抗体在裸鼠体内显著抑制naLAT 1/3 T3细胞的体外细胞增殖和体内肿瘤生长，表明LAT 1是一个有前途的抗癌靶点。
L-type amino acid transporter 1 (LAT1)/SLC7A5 is the first identified CD98 light chain disulfide linked to the CD98 heavy chain (CD98hc/SLC3A2). LAT1 transports large neutral amino acids, including leucine, which activates mTOR, and is highly expressed in human cancers. We investigated the oncogenicity of human LAT1 introduced to NIH/3T3 cells by retrovirus infection. NIH/3T3 cell lines stably expressing human native (164C) or mutant (164S) LAT1 (naLAT1/3T3 or muLAT1/3T3, respectively) were established. We confirmed that endogenous mouse CD98hc forms a disulfide bond with exogenous human LAT1 in naLAT1/3T3, but not in muLAT1/3T3. Endogenous mouse CD98hc mRNA increased in both naNIH/3T3 and muLAT1/3T3, and a similar amount of exogenous human LAT1 protein was detected in both cell lines. Furthermore, naLAT1/3T3 and muLAT1/3T3 cell lines were evaluated for cell growth-related phenotypes (phosphorylation of ERK, cell-cycle progression) and cell malignancy-related phenotypes (anchorage-independent cell growth, tumor formation in nude mice). naLAT1/3T3 had stronger growth- and malignancy- related phenotypes than NIH/3T3 and muLAT1/3T3, suggesting the oncogenicity of native LAT1 through its interaction with CD98hc. Anti-LAT1 monoclonal antibodies significantly inhibited in vitro cell proliferation and in vivo tumor growth of naLAT1/3T3 cells in nude mice, demonstrating LAT1 to be a promising anti-cancer target.