人表皮生长因子受体显性负性突变体对胃癌细胞体外侵袭转移能力的抑制作用

Inhibitive effect of dominant negative human epidermal growth factor receptor on the invasion and metastasis of gastric cancer cells

摘要:

目的：探讨人表皮生长因子受体（Epidermal growth factor receptor，EGFR）显性负性突变体（Dominant negative epidermal growth factor receptor，DNEGFR）对胃癌细胞体外侵袭转移能力的影响及其分子机制。方法：构建pEGFPN1-DNEGFR，转染人胃癌细胞并筛选稳定转染株，采用Transwell侵袭小室模型检测人胃癌细胞体外侵袭转移能力，RT-PCR检测细胞基质金属蛋白酶-2（Matrix metalloproteinase-2，MMP-2）和基质金属蛋白酶-9（Matrix metalloproteinase-9，MMP-9）mRNA水平，酶联免疫吸附测定（Enzyme-linked immunosorbent assay，ELISA）检测培养液MMP-2和MMP-9 蛋白水平。结果：成功构建pEGFPN1-DNEGFR，转染并筛选出稳定转染株，检测到人胃癌细胞体外侵袭转移能力降低，细胞MMP-2和MMP-9 mRNA水平降低，培养液MMP-2和MMP-9 蛋白水平降低。结论：DNEGFR通过抑制人胃癌细胞分泌MMP-2和MMP-9，使其体外侵袭转移能力显著降低，将为DNEGFR在胃癌生物治疗中的深入研究打下基础。

Abstract:

Objective：To explore the effect of dominant negative epidermal growth factor receptor（DNEGFR） on the invasion and metastasis of human gastric cancer cells and to elucidate its molecular mechanism. Methods：The pEGFPN1-DNEGFR was construct－ed and human gastric cancer cells were transfected stably. The invasion and metastasis in vitro were tested by Transwell invasion cham－ber assay. Matrix metalloproteinase-2（MMP-2） and Matrix metalloproteinase-9（MMP-9） mRNA levels of cells were detected by re－versed transcription-polymerase chain reaction（RT-PCR）. MMP-2 and MMP-9 protein levels of culture were detected by enzyme-linked immunosorbent assay（ELISA）. Results：pEGFPN1-DNEGFR was constructed successfully，human gastric cancer cells was transfected stably. MMP-2 and MMP-9 mRNA levels of cells were down-regulated；MMP-2 and MMP-9 protein levels of culture were down-regulated as well. Conclusions：DNEGFR represses the invasion and metastasis of human gastric cancer cells by inhibiting them from secreting MMP-2 and MMP-9，which lays a solid foundation for further research of DNEGFR in gastric cancer biotherapy.