Antitumor effects of novel mAbs against cationic amino acid transporter 1 (CAT1) on human CRC with amplified CAT1 gene.
Antitumor effects of novel mAbs against cationic amino acid transporter 1 (CAT1) on human CRC with amplified CAT1 gene.
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DOI：
10.1111/cas.14741
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发表时间：
2021-03
期刊：
影响因子：
5.7
通讯作者：
Masuko T
中科院分区：
文献类型：
--
作者：
Okita K;Hara Y;Okura H;Hayashi H;Sasaki Y;Masuko S;Kitadai E;Masuko K;Yoshimoto S;Hayashi N;Sugiura R;Endo Y;Okazaki S;Arai S;Yoshioka T;Matsumoto T;Makino Y;Komiyama H;Sakamoto K;Masuko T
通过比较基因组杂交(CGH)检测到的拷贝数改变可以导致新的癌症相关基因的识别。我们用计算全息技术分析了100例人类原发结直肠癌(CRC)的染色体异常，发现了一个溶质载体(SLC)7A1基因，它编码阳离子氨基酸转运蛋白1(CAT1)，有14个跨膜区，位于染色体(13q12.3)上，基因扩增频率较高。SLC7A1/CAT1是一种转运蛋白，负责摄取细胞生长所必需的阳离子氨基酸(精氨酸、赖氨酸和鸟氨酸)。基因芯片和聚合酶链式反应分析表明，从CAT1转录的mRNA在70%以上的人结直肠癌组织中过表达，RNA干扰介导的CAT1基因敲除抑制了癌细胞的生长。用绿色荧光蛋白(GFP)标记的表达CAT1的大鼠肝癌细胞免疫大鼠，并将大鼠脾细胞与小鼠骨髓瘤细胞融合。从已建立的杂交瘤克隆中筛选出5株与表达CAT1-GFP的HEK293细胞以GFP表达依赖的方式反应的大鼠单抗(CA1~CA5)。免疫组织化学结果显示，新型抗CAT1单抗与人结直肠癌组织及邻近正常组织有选择性反应，流式细胞仪检测发现其与多种人癌细胞株有较强的结合。抗CAT1单抗对结直肠癌细胞具有内化活性、抗体依赖的细胞毒作用和迁移抑制活性。此外，CA2还能抑制裸鼠体内人HT29和Sw-C4结直肠癌的生长。本研究提示CAT1有望成为抗癌单抗治疗的靶点。在结直肠癌中，SLC7A1/CAT1基因扩增频繁，CAT1基因表达增加。研制了新型抗CAT1单抗，证实了CAT1蛋白在结直肠癌中的高表达及单抗对裸鼠移植瘤的体内抗肿瘤作用。CAT1可能是抗结直肠癌治疗的一个有前景的靶点。
Copy number alterations detected by comparative genomic hybridization (CGH) can lead to the identification of novel cancer‐related genes. We analyzed chromosomal aberrations in a set of 100 human primary colorectal cancers (CRCs) using CGH and found a solute carrier (SLC) 7A1 gene, which encodes cationic amino acid transporter 1 (CAT1) with 14 putative transmembrane domains, in a chromosome region (13q12.3) with a high frequency of gene amplifications. SLC7A1/CAT1 is a transporter responsible for the uptake of cationic amino acids (arginine, lysine, and ornithine) essential for cellular growth. Microarray and PCR analyses have revealed that mRNA transcribed from CAT1 is overexpressed in more than 70% of human CRC samples, and RNA interference–mediated knockdown of CAT1 inhibited the cell growth of CRCs. Rats were immunized with rat hepatoma cells expressing CAT1 tagged with green fluorescent protein (GFP), and rat splenocytes were fused with mouse myeloma cells. Five rat monoclonal antibodies (mAbs) (CA1 ~ CA5) reacting with HEK293 cells expressing CAT1‐GFP in a GFP expression–dependent manner were selected from established hybridoma clones. Novel anti‐CAT1 mAbs selectively reacted with human CRC tumor tissues compared with adjacent normal tissues according to immuno‐histochemical staining and bound strongly to numerous human cancer cell lines by flow cytometry. Anti‐CAT1 mAbs exhibited internalization activity, antibody‐dependent cellular cytotoxicity, and migration inhibition activity against CRC cell lines. Furthermore, CA2 inhibited the in vivo growth of human HT29 and SW‐C4 CRC tumors in nude mice. This study suggested CAT1 to be a promising target for mAb therapy against CRCs. Frequent gene amplification of SLC7A1/CAT1 and increased CAT1 mRNA expression in colorectal cancers (CRC) were demonstrated. Novel anti‐CAT1 monoclonal antibodies (mAbs) were developed, and higher expression of CAT1 protein in CRC and in vivo antitumor effects of mAbs on xenografted CRC cells in nude mice were confirmed. CAT1 may be a promising target for anti‐CRC therapy.
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影响因子：
158.5
作者：
Lynch, TJ;Bell, DW;Haber, DA
通讯作者：
Haber, DA
影响因子：
8.8
作者：
Kremer JC;Prudner BC;Lange SES;Bean GR;Schultze MB;Brashears CB;Radyk MD;Redlich N;Tzeng SC;Kami K;Shelton L;Li A;Morgan Z;Bomalaski JS;Tsukamoto T;McConathy J;Michel LS;Held JM;Van Tine BA
通讯作者：
Van Tine BA
影响因子：
6.2
作者：
Dillon, BJ;Prieto, VG;Clark, MA
通讯作者：
Clark, MA
影响因子：
4.2
作者：
Closs, EI;Simon, A;Rotmann, A
通讯作者：
Rotmann, A
影响因子：
20.3
作者：
Miraki-Moud, Farideh;Ghazaly, Essam;Taussig, David C.
通讯作者：
Taussig, David C.