摘要
[目的]研究pPeOp蛋白对胃癌细胞MC-4迁移的抑制作用,为雷丸的开发应用奠定理论基础。[方法]通过MTT法研究p Pe Op蛋白对人胃癌细胞MC-4的增殖抑制作用;划痕实验观察pPeOp蛋白干预后MC-4的迁移情况;RT-PCR、明胶酶谱法以及ELISA法分析MMP-2和MMP-9的表达和活性。[结果]在30μg·m L^-1-120μg·m L^-1浓度范围内,随着pPeOp浓度的增加,MC-4细胞的存活率逐渐减小;划痕实验显示MC-4细胞的迁移能力受到不同程度的抑制,且在p Pe Op蛋白浓度为120μg·m L^-1时,抑制作用最明显;RT-PCR的结果显示,MC-4细胞在对照组和30μg·m L^-1p Pe Op蛋白组中能检测到MMP-2基因的表达产物,但在45μg·m L^-1-120μg·m L^-1浓度范围内未检测到MMP-2基因的表达产物;明胶酶谱法结果表明MMP-2含量随pPeOp浓度的增加而减小;各组均无法检测到MMP-9蛋白的条带;ELISA法结果显示,pPeOp蛋白可降低MC-4细胞MMP-2的分泌,但对MMP-9无显著影响。[结论]pPeOp蛋白能够抑制胃癌细胞MC-4的增殖和迁移,且迁移抑制作用与MMP-2具有一定的相关性,与MMP-9无显著相关性,具体作用机制尚需进一步深入研究。
[Objective] Evaluate the migration activity of pPeOp on human gastric tumor cell in vitro and provide a meaningful basis for early diagnosis and treatment. [Methods]MTT assay and wound scratch assay were utilized to estimate the cell proliferation and migration ability. The m RNA expression of metallopeptidase 2（MMP-2） and MMP-9 was observed by RT-PCR. Then the activity of MMP-2 and MMP-9 was measured by gelatin zymography and ELISA assay. [Results]With the pPeOp protein concentration increasing,the survival rate of MC-4 cell declined significantly. Wound scratch assay showed that the pPeOp protein possessed a significant migration inhibition on MC-4 cell in a concentration-dependent manner,with a maximum inhibition at the highest concentration. RT-PCR stated that MMP-2 expressed only in the control group and the lowest concentration of pPeOp protein.Zymography assay demonstrated MMP-2 protein activity decreased with increasing concentration of pPeOp. However,MMP-9 was not expressed in any treatment group. ELISA assay showed that pPeOp can reduce the MMP-2 activity of MC-4 cells,but had no effect on MMP-9. [Conclusion]The paper proved that the pPeOp protein of O. lapidescens can inhibit the activity and migration of gastric cancer cells. The inhibition had so me relevance with MMP-2, but not with MMP. It needs further study to explain the mechanism.
出处
《浙江中医药大学学报》
CAS
2015年第1期9-14,共6页
Journal of Zhejiang Chinese Medical University
基金
国家自然科学基金(81374023)
浙江省自然科学基金(Y207765)
浙江省大学生科技创新活动计划(2013R410029)~~
