Human Recombinant Arginase I [HuArgI (Co)-PEG5000]-Induced Arginine Depletion Inhibits Colorectal Cancer Cell Migration and Invasion
Human Recombinant Arginase I [HuArgI (Co)-PEG5000]-Induced Arginine Depletion Inhibits Colorectal Cancer Cell Migration and Invasion
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DOI：
10.3390/ijms20236018
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发表时间：
2019-12-01
影响因子：
5.6
通讯作者：
El-Sibai, Mirvat
中科院分区：
文献类型：
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作者：
Al-Koussa, Houssam;Al-Haddad, Maria;El-Sibai, Mirvat
结直肠癌（CRC）是全球第三大常见癌症，占所有胃肠道癌症死亡的一半以上。已知癌细胞具有高增殖率，它们需要大量的氨基酸，包括精氨酸。此外，几种肿瘤类型已被证明下调ASS-1表达，成为精氨酸营养缺陷型。因此，精氨酸剥夺是靶向癌细胞的有前途的治疗方法之一。这可以通过使用重组人精氨酸降解酶HuArgI（Co）-PEG 5000来实现。研究方法：在本研究中，通过细胞毒性测定来检查HuArgI（Co）-PEG 5000对CRC细胞系（HT-29、Caco-2、Sw 837）的细胞毒性作用。还进行伤口愈合测定、侵袭测定和粘附测定以检测对转移的影响。结果：伤口愈合和侵袭试验显示，用酶处理后细胞迁移和侵袭减少。用荧光素酶处理的细胞也显示出粘附的减少，这与RhoA活化的减少一致，这通过使用FRET生物传感器在单细胞测定中检测RhoA活化和MMP-9表达的减少来证明。用精氨酸酶和L-瓜氨酸处理细胞，可显著恢复细胞内精氨酸水平，逆转HuArgI（Co）-PEG 5000对细胞活力、迁移和侵袭的影响。结论：因此，我们可以得出结论，结肠直肠癌是部分营养缺陷型精氨酸和精氨酸耗尽是一个潜在的选择性抑制结肠癌细胞的运动和侵袭的方法。
Purpose: Colorectal cancer (CRC) is the third most common type of cancer worldwide, and it represents over half of all gastrointestinal cancer deaths. Knowing that cancer cells have a high proliferation rate, they require high amounts of amino acids, including arginine. In addition, several tumor types have been shown to downregulate ASS-1 expression, becoming auxotrophic for arginine. Therefore, Arginine deprivation is one of the promising therapeutic approaches to target cancer cells. This can be achieved through the use of a recombinant human arginase, HuArgI(Co)-PEG5000, an arginine degrading enzyme. Methods: In this present study, the cytotoxic effect of HuArgI(Co)-PEG5000 on CRC cell lines (HT-29, Caco-2, Sw837) is examined though cytotoxicity assays. Wound healing assays, invasion assays, and adhesion assays were also performed to detect the effect on metastasis. Results: Wound healing and invasion assays revealed a decrease in cell migration and invasion after treatment with arginase. Cells that were treated with arginase also showed a decrease in adhesion, which coincided with a decrease in RhoA activation, demonstrated though the use of a FRET biosensor to detect RhoA activation in a single cell assay, and a decrease in MMP-9 expression. Treating cells with both arginase and L-citrulline, which significantly restores intracellular arginine levels, reversed the effect of HuArgI(Co)-PEG5000 on cell viability, migration, and invasion. Conclusion: We can, therefore, conclude that colorectal cancer is partially auxotrophic to arginine and that arginine depletion is a potential selective inhibitory approach for motility and invasion in colon cancer cells.