喹诺酮类化合物H25抗肿瘤侵袭活性体外研究  

Inhibition of a new quinolone derivative H25 on cancer invasion in vitro

作　　者：贺晓波[1] 刘伟[1] 姜威[1] 尤启冬[2] 司书毅[1] 

机构地区：[1]中国医学科学院医药生物技术研究所,北京100050 [2]中国药科大学药学院,南京210009

出　　处：《中国抗生素杂志》2010年第5期391-394,共4页Chinese Journal of Antibiotics

基　　金：国家自然科学基金(30271543);"重大新药创制"科技重大专项(2009ZX09501-003)资助

摘　　要：目的研究喹诺酮类化合物H25抗肿瘤细胞侵袭转移的活性,初步探讨其作用机制。方法明胶酶谱法验证H25对Ⅳ型胶原酶的竞争性抑制作用。MTT法检测H25对HT1080细胞的生长抑制作用。TranswellTM侵袭小室试验检测H25对HT1080细胞侵袭转移的抑制作用。明胶酶谱法和RT-PCR法检测H25对MMP-2、MMP-9表达水平的影响。细胞粘附试验观察H25对HT1080细胞粘附作用的影响。结果 H25能竞争性抑制Ⅳ型胶原酶的水解酶活性。5h和48h药物处理条件下,H25对HT1080细胞生长抑制作用的IC50分别为13.51和1.311μmol/L。TranswellTM侵袭小室试验中,H25对HT1080细胞侵袭Matrigel的抑制率分别为49.1%(1μmol/L,5h)、25.9%(0.1μmol/L,5h)、69.4%(0.1μmol/L,48h)、39.6%(0.01μmol/L,48h),其有效抑制浓度低于细胞生长抑制浓度。明胶酶谱分析证明,H25处理过的HT1080细胞培养上清中Ⅳ型胶原酶活性明显降低。但RT-PCR分析未检测到MMP-2、MMP-9mRNA表达水平的变化。另外,1μmol/L的H25能抑制HT1080细胞的异质粘附作用。结论 H25能有效抑制HT1080细胞的侵袭,不仅因为H25是Ⅳ型胶原酶的竞争性抑制剂,而且可能与其对MMP-2、MMP-9表达的转录后抑制和异质粘附抑制相关。Objective A new quinolone derivative H25, gotten in screening for Type Ⅳ collagenase （Col. Ⅳ） inhibitor, was investigated in this study about its effect on invasion and metastasis of cancer cells and the possible molecular mechanism. Methods Gelatin zymography assay was employed to verify the competitive inhibitory effect of H25 on Col.Ⅳ. MTT assay was used to value growth inhibition of HT1080 cell treated with H25. The Transwell^TM system was used for invasion and migration assays. The expression of MMP-2 and MMP-9 was analyzed by zymography assay and RT-PCR assay. Cell adhesion assay was used to investigate the effect of H25 on adhesion character of HT1080 cell. Results Experimental result verified the ability of H25 to competitively inhibit Col. IV. H25 showed cytostatic effect on HT1080 cell with IC50 of 13.51μmol/L and 1.31μmol/L respectively after 5h and 48h treatment. H25 inhibited HT1080 cell to invade through Matrigel layor with inhibition ratio of 49. 1% （1 μmol/ L for 5h）, 25.9% （0.1μmol/L for 5h）, 69.4% （0.1μmol/L for 48h）, 39.6% （0.01μmol/L for 48h）, and the inhibition occurred without affecting cell proliferation. Type Ⅳ collagenase activity in the serum-free supernatant of HT1080 cells treated with H25 was reduced, but no change was observed on expression of MMP-2 and MMP-9 mRNA by RT-RCR assay. In addition, H25 could inhibit heterogeneous adhesion of HT1080 cell effectually at 1 μmol/L. Conclusion H25 could inhibit cell invasion in vitro, which was not only caused by its Col. Ⅳ inhibitor activity, but also possibly induced by Col. Ⅳ expression inhibition in post-transcription level and heterogeneous adhesion inhibition.

关 键 词：喹诺酮 Ⅳ型胶原酶 抑制剂 HT1080细胞 

分 类 号：R978.1[医药卫生—药品]