摘要
目的:通过体外实验研究钠氢交换蛋白1(NHE1)特异性抑制剂Cariporide对人宫颈癌HeLa细胞体外转移能力的影响,并初步探讨其作用机制。方法:用Cariporide处理HeLa细胞,采用细胞-基质粘附实验、细胞划痕实验和Transwell侵袭实验分别检测Cariporide对HeLa细胞的粘附、迁移和侵袭能力的影响:Real-time RT-PCR、Western blot法分别检测Cariporide对HeLa细胞膜型基质金属蛋白酶1(MT1-MMP)mRNA及蛋白表达的影响结果:细胞-基质粘附实验结果显示,与对照组相比,Cariporide处理过的HeLa细胞体外粘附率降低25%,差异具有统计学意义(P<0.05);细胞划痕实验和Transwell侵袭实验结果显示,Cariporide处理后,HeLa细胞损伤愈合的速度和穿过滤膜的细胞数量均明显少于对照组;Real-time RT-PCR及Western blot结果表明Cariporide可显著降低MT1-MMPmRNA和蛋白水平的表达量(P均<0.05)。结论:Catiporide可有效抑制人宫颈癌HeLa细胞的体外转移能力,其作用机制可能与降低MT1-MMP的表达有关。
Objective: To investigate the effects of sodium hydrogen exchange protein 1 (Na+/H+ exchanger 1; NHE 1)-specific in- hibitor Cariporide on the in vitro metastasis of HeLa cells and to preliminarily explore the potential anti-metastasis mechanism of Cariporide. Methods: HeLa cells were treated with Cariporide, and its effects on the adhesion, migration, and invasion of HeLa cells were determined using cell-matrix adhesion assay, wound healing assay, and cell invasion assay. Real-time RT-PCR and Western blot assay were used to detect the effects of Cariporide on the levels of MT1-MMP ( membrane-type 1 matrix metalloproteinase ) mRNA and on the protein expression in the Hela cells, respectively. Results: The cell-matrix adhesion assay showed that the adhesive ratio of the HeLa cells treated with Cariporide was reduced by 25% compared with the controls. There were also significant differences between the two groups ( P〈 0.05 ). The wound healing and cell invasion assays showed that after the Cariporide treatment, the healing speed of HeLa cell injury and the amount of the cells passing through the filter membrane were less in the treatment groups than in the control groups. RT-PCR and Western blot results showed that Cariporide produced a distinguished inhibition of MT1-MMP expression in HeLa cells, both on the mRNA and on the protein levels ( P 〈 0.05 ). Conclusion: Cariporide can inhibit the in vitro metastasis of HeLa cells effectively. The mechanism may be related to the downregulation of the expression of MT 1-MMP.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2011年第17期1028-1031,共4页
Chinese Journal of Clinical Oncology
基金
教育部高等学校博士学科点专项科研基金(编号:20091106110038)
天津市自然科学基金重点项目(编号:09JCZDJC17300)资助~~
关键词
钠氢交换蛋白1
膜型基质金属蛋白酶1
粘附
迁移
侵袭
Na+/H+ exchanger 1
Membrane-type 1 matrix metalloproteinase
Adhesion
Migration
Invasion