胃泌素对人胃癌细胞裸鼠移植瘤生长与转移潜能影响研究  

Effects of gastrin over-expression on the growth of xenografts in nude mice and the metastasis of gastric cancer cells

作　　者：曾怡[1] 姜琼[2] 周建奖[1] 谢渊[1] 赵艳[1] 

机构地区：[1]贵阳医学院分子生物学重点实验室,贵州贵阳550004 [2]贵阳医学院附属医院内镜中心,贵州贵阳550004

出　　处：《中华肿瘤防治杂志》2015年第19期1505-1511,共7页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81260303);贵州省科技计划基金(黔科合SY字[2011]3067号)

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。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 abil- ity in vivo and the cell invasion and metastasis in vitro were detected by nude mice xenograft, cell wound healing assay and Tran- swell 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 gastrin- overexpressed cells and higher than that in control ceils （50%）. Twenty-five days after inoculation, the xenograh 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 ± 0. 352 6） cm3 and （0. 128 0±0. 201 4） g in control cells, and P values 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 X field （SGC-7901） and （86±5） and （24±4） cells/400 Xfield （ACTS） 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/400X field, significantly higher than the （16±5） cells and （68±3） cells/400 X 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 MMP2 and VEGF protein increased by 166% and 190% （SGC-7901） and 50o/oo and 68% （AGS）, as well as the rela- tive expression of MMP2 and VEGF mRNA was up-regulated by 20.82 and 98.61 times （SGC-7901） and 32. 58 and 26. 71 times （AC

关 键 词：胃肿瘤 胃泌素 裸鼠成瘤 肿瘤转移 肿瘤浸润 印迹法 蛋白质 

分 类 号：R735.2[医药卫生—肿瘤]