摘要
目的研究S100A11表达下调对胃癌细胞增殖、细胞周期和侵袭的影响。方法采用Western blot检测不同胃癌细胞株(MKN-28、SGC-7901和MKN-45)和正常胃黏膜上皮细胞(GES-1)中S100A11蛋白的表达。将S100A11siRNA和对照siRNA分别转染MKN-45细胞,将细胞分为3组:未处理组、对照siRNA组和S100A11 siRNA组。采用Western blot检测转染前后S100A11蛋白的表达,并利用CCK-8试剂、流式细胞术及Boyden小室分别检测3组细胞的增殖、细胞周期和细胞侵袭能力的变化,最后采用Western blot分析细胞周期和侵袭相关蛋白的表达。结果 S100A11蛋白在胃癌组织中表达的阳性率(69.81%,37/53)明显高于正常胃黏膜组织(16.98%,9/53)(χ2=30.110,P=0.000)。3株胃癌细胞株中S100A11蛋白表达水平显著高于正常胃黏膜上皮细胞(P<0.05),其中S100A11在MKN-45细胞中的表达水平显著高于MKN-28和SGC-7901(P<0.05)。S100A11 siRNA能显著下调MKN-45细胞中S100A11蛋白的水平,其表达下调显著抑制细胞的增殖、改变细胞周期分布和降低细胞的侵袭能力。S100A11表达下调能显著降低Cyclin D1、Cdk2和MMP-2蛋白的表达,但增加E-cadherin蛋白的表达水平。结论 S100A11介导胃癌细胞生物学行为的变化可能与Cyclin D1、Cdk2、E-cadherin和MMP-2表达变化密切相关,因而抑制S100A11的水平可能是胃癌潜在的分子治疗靶点。
Objective To investigate the effects of S100All expression down-regulation on the proliferation, cell cycle and invasion in gastric carcinoma cells. Methods Expression of S100A11 protein was examined in 3 gastric carcinoma cell lines （MKN-28, SGC-7901 and MKN-45 ） and normal gastric mucous epithelial cell GES-1 by Western blotting. S100A11 siRNA and control siRNA were utilized to transfect MKN- 45 cells, which were divided into 3 groups, untreated group, control siRNA group and S100A11 siRNA group. Expression of S100A11 protein was investigated after transfeetion by Western blotting. Subsequently, the chan- ges of cell proliferation, cell cycle and cell invasion were detected by CCK-8 kit, flow cytometry and Boyden chamber assay, respectively. Finally, the expressions of cell cycle- and invasion-related proteins were analyzed by Western blotting. Results Positive expression ratio of S100A11 in gastric carcinoma tissues was significantly higher than that in normal gastric mucous epithelial tissues （ Chi square = 30. 110, P = 0. 000 ）. The levels of S100A11 protein expression in the 3 gastric carcinoma cell lines were significantly higher than that in normal gastric mucous epithelial cells （P 〈 0. 05 ）. In addition, expression of S100A11 protein in MKN-45 cells was markedly higher than that in MKN-28 and SGC-7901 cells （P 〈 0.05 ）. S100A11 siRNA significantly downreg- ulated the level of S100A11 protein in MKN-45 cells, and the downregulation evidendy suppressed cell proliferation, altered cell cycle distribution and reduced cell invasion ability. Most notably, the downregulation of S100A11 expression significantly evoked the expression decrease of Cyclin D1, Cdk2 and MMP-2 and increase of E-cadherin. Conclusion S100All mediates the alterations of biological behaviors of gastric carcinoma cells, which may be tightly associated with the expression changes of Cyclin DI, Cdk2, E-cadherin and MMP-2. Thus, S100AI 1 may be a potential molecular therapy target of gastric carcinoma.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2013年第24期2665-2670,共6页
Journal of Third Military Medical University
