α-苦瓜素和PEG-α-苦瓜素抑制绒癌JAR细胞迁移和侵袭的研究

Inhibitory effect of α-momorcharin and its PEGylated conjugates on the migration and invasion of human choriocarcinoma JAR cells

目的考察α-苦瓜素(α-MMC)对绒癌JAR细胞体外迁移侵袭行为的影响及其聚乙二醇(PEG)修饰物保留这一活性的程度。方法利用分子量为20 kDa的聚乙二醇氨基酸衍生物[(mPEG)2-Lys-NHS]以共价连接的方式对α-MMC分子表面进行修饰。采用MTT法、细胞黏附试验、划痕损伤试验、transwell法和明胶酶谱法分别检测JAR细胞经α-MMC或PEG-α-MMC作用后细胞的生长、存活、黏附、迁移、侵袭行为及分泌基质金属蛋白酶(MMPs)能力的变化。结果α-MMC及PEG-α-MMC依赖于剂量和时间方式显著抑制了绒癌JAR细胞的生长,PEG-α-MMC可保留67%天然α-MMC的抗增殖活性(1.5 mg.mL-1作用72 h)。经修饰后蛋白处理,细胞分泌的基质金属蛋白酶-2(MMPs-2)活性减弱,削弱了细胞的运动能力,JAR细胞的行进过程受到抑制。结论α-MMC及其PEG修饰物通过减弱MMPs-2活性抑制绒癌细胞的体外行进过程,首次揭示了核糖体失活蛋白可能影响体外肿瘤转移过程。

OBJECTIVE To assess the anti-invasive ability of α-momorcharin（α-MMC） and evaluate to the PEGylated modefication of α-MMC to maintain the activity.METHODS The surface modification was performed with a 20 kDa amino-derivative PEG（（mPEG）2-Lys-NHS）to α-MMC by covalent attachment.The quantitative MTT test,adhesion assay,wound healing assay,matrigel invasion assay and gelatin zymography were carried out to test the cell viability and proliferation,cell adhesion,migration and invasiveness,and the activities of matrix metalloproteinases（MMPs）of JAR cells after proteins treatment.RESULTS Both native and PEGlyated α-MMC significantly inhibited the growth of choriocarcinoma JAR cells in a dose-and time-dependent manner.PEG-α-MMC preserved about 67% antiproliferative activity of native（1.5 mg·mL-1,treatment for 72 h）.The activity of matrix-metalloproteinase MMPs-2 was potently reduced after protein treatment,which was associated with decreasing cell movement potential.In vitro cell migration and invasion assays showed the coincident results.CONCLUSION The inhibitory of α-MMC and its PEGylated conjugates on the metastatic stage of human choriocarcinoma JAR cells indicates the possible role of RIP in tumor progression.