【摘要】 目的探讨胃泌素在胃癌细胞生长、侵袭和转移中的作用。方法构建过表达胃泌素的细胞模型,用裸鼠成瘤实验检测体内肿瘤形成;细胞伤愈和Transwell小室实验检测体外细胞侵袭和转移;蛋白质印迹法、实时荧光定量PCR和酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)检测基质金属蛋白酶-2(matrix metalloproteinases,MMP-2)和血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达及分泌。结果过表达胃泌素的胃癌细胞,裸鼠体内成瘤率为100%(6/6),高于对照细胞50%(3/6)。接种后25d,移植瘤的体积和体质量分别为(0.980 9±0.626 0)cm3和(0.445 6±0.280 1)g,均高于对照细胞的(0.222 1±0.352 6)cm3和(0.128 0±0.201 4)g,P值分别为0.018和0.048。Transwell小室实验结果显示,过表达胃泌素和对照的SGC-7901细胞的迁移细胞数为(24±4)和(10±5)个/400倍视野,q=4.24,P<0.05;AGS细胞为(86±5)和(24±4)个/400倍视野,q=4.93,P<0.05。SGC-7901细胞的侵袭细胞数为(58±5)和(16±5)个/400倍视野,q=4.65,P<0.05;AGS细胞为(112±5)和(68±3)个/400倍视野,q=5.24,P<0.05。蛋白质印迹法、实时荧光定量PCR和ELISA结果显示,与对照细胞比较,过表达胃泌素的SGC-7901细胞中MMP-2和VEGF蛋白相对表达量分别增加166%和190%,AGS细胞中分别增加50%和68%;SGC-7901细胞中MMP-2和VEGF mRNA的相对表达量分别上调20.82倍和98.61倍,AGS细胞中分别上调32.58倍和26.71倍;培养液中MMP-2和VEGF的含量也增加,差异均有统计学意义,P值均<0.05。加入丙谷胺后,胃泌素的促进作用被抑制。结论胃泌素能促进胃癌细胞体内外肿瘤生长、侵袭和转移,有可能成为阻断胃癌侵袭转移的新靶点。更多还原

【Abstract】 OBJECTIVE To study the effects of gastrin on the growth,invasion and metastasis of gastric cancer cells.METHODS Human gastric cancer SGC-7901 and AGS cells with gastrin over-expression were established.Tumor formation ability in vivo and the cell invasion and metastasis in vitro were detected by nude mice xenograft,cell wound healing assay and Transwell assay.The expression level of matrix metalloproteinases-2(MMP-2)and vascular endothelial growth factor(VEGF)mRNA and protein and their concentrations in culture medium were measured by Real-time RT-PCR,Western blot and Enzyme-Linked Immunosorbent Assay(ELISA)respectively.RESULTS The tumor formation rate in nude mice was 100%(6/6)in gastrinoverexpressed cells and higher than that in control cells(50%).Twenty-five days after inoculation,the xenograft volume and weigh in experiment cells were(0.980 9±0.626 0)cm3 and(0.445 6±0.280 1)g,which was higher than the(0.222 1±0.352 6)cm3 and(0.128 0±0.201 4)g in control cells,and Pvalues were 0.018 and 0.048,respectively.Transwell assay showed that the numbers of migrating cells in gastrin-overexpression and control cells were(24±4)cells and(10±5)cells/400×field(SGC-7901)and(86±5)and(24±4)cells/400×field(AGS)respectively,with a significant difference between them(all P<0.05).The numbers of invading SGC-7901 and AGS cells in gastrin-overexpressed groups were(58±5)cells and(112±5)cells/400×field,significantly higher than the(16±5)cells and(68±3)cells/400×field in control group(all P<0.05).Western blot,Real-time RT-PCR and ELISA assays revealed that compared with the control cells,the relative expression levels of MMP-2and VEGF protein increased by 166%and 190%(SGC-7901)and 50%and 68%(AGS),as well as the relative expression of MMP-2and VEGF mRNA was up-regulated by 20.82 and 98.61times(SGC-7901)and 32.58 and 26.71times(AGS)respectively in gastrin-overexpressed cells(all P<0.05).Further the contents of MMP-2and VEGF in culture medium in gastrin-overexpressed cells were increased(P<0.05).After proglumid treatment,the gastrin promoting ability was inhibited.CONCLUSION Gastrin could promote the tumor growth,invasion and metastasis of gastric cancer cells in vivo and vitro,which may become a new target for blocking the invasion and metastasis of gastric cancer.更多还原