摘要
目的 体外考察趋化因子受体7 （chemokine receptor 7,CXCR7）在胃癌细胞MGC-803中的表达和CXCR7-siRNA重组腺病毒表达载体靶向抑制CXCR7的表达对人胃癌细胞MGC-803增殖、迁移活性的影响.方法 将MGC-803细胞分为实验组、空载体组及空白对照组,实验组加入CXCR7-siRNA重组腺病毒,空载体组加入空载腺病毒,空白对照组加入等量容积细胞培养基.噻唑蓝（MTT）法测各组细胞增殖活性;Hoechst 33258 染色法观察各组细胞凋亡情况;Western blot 免疫印迹法检测各组MGC-803细胞中CXCR7的表达情况及凋亡执行蛋白Caspase-3 表达情况; Transwell细胞迁移实验检测各组细胞趋化活性.结果 实验组细胞增殖受到抑制,转染3天后,实验组细胞MTT值为1.22±0.01,与空载体转染组（1.48±0.05）及空白对照组（1.61±0.03）比较,差异有统计学意义（F=31.2,P＜0.05）.实验组细胞凋亡比率为（37.66±1.31）%,与空载体组（3.39±0.76）%及空白对照组（13.12±0.85）%比较差异具有统计学意义（F=391.9,P〈0.05）.实验组CXCR7表达水平低于空载体组和空白对照组;实验组Caspase-3剪切激活,空载体组见少许Caspase-3剪切激活,空白对照组未见Caspase-3剪切激活;转染3天后,Transwell细胞迁移实验显示实验组穿透滤过膜细胞数量（15.00±1.73）,较空载体组（35±2.89）及空白对照组（37±4.04）显著减少,差异均有统计学意义（F=12.51,P〈0.05）.结论 靶向沉默CXCR7的表达抑制胃癌细胞MGC-803的增殖和迁移活性.
Objective To investigate the expression of chemokine receptor 7 （ CXCR7 ） in human gastric cancer MGC - 803 cells and changes in the proliferation and migration of MGC - 803 cells after inhibition of CXCR7 by recombi- nant adenovirus CXCR7 - siRNA. Methods MGC - 803 cells were divided into a treatment group, an empty vector group and a control group, which received recombinant adenovirus CXCR7 -siRNA, empty vectors or culture media re- spectively. Then cell proliferation was measured by MTF assay. The apoptotie effects in each group were detected by Ho- echst 33258 staining. The expressions of CXCR7 and caspase - 3 were examined by Western blotting. The chemotactic activities were determined using Transwell migration assay. Results The proliferation of MGC - 803 cells in the treat- ment group was remarkably inhibited. Three days after transfection, the MTr absorbance value in the treatment group were （1.22 ±0.01 ）, which was significantly statistic different from those in the empty vector group （1.48 ±0.05 ） and the control group （ 1.61 ±0.03 ） （ F = 31.2, P 〈 0.01 ）. The apoptotic ratio of the treatment group was （ 37.66 ±1.31 ） % , which was significantly different from those in the empty vector group （3.39 ±0.76） % and the control group （ 13.12 ±0.85 ） % （ F = 391.9, P 〈0.05 ）. Western blotting analysis showed a lower level of CXCR7 in the treatment than in either the empty vector or control group. Activated caspase - 3 was detected in the treatment group while less was seen in the empty vector group and none in the control group. Three days after transfection, Transwell migration assay re- vealed that the number of MGC - 803 cells penetrating the filtration membrane in the treatment group was （ 15.00 ±1.73 ）, which was significantly lower than those in the empty vector group （ 35 ±2.89 ） （ P 〈 0.05 ） and the control group （37 ±4.04） （ P 〈 0.05 ）. Conclusion Targeti^g inhibition of CXCR7 expression can suppress the proliferative and migratory activities of gastric cancer MGC - 803 cells.
出处
《徐州医学院学报》
CAS
2013年第11期717-720,共4页
Acta Academiae Medicinae Xuzhou
基金
江苏省肿瘤生物治疗重点实验室开放课题（ZL1206）
