钙离子结合蛋白S100A16对胃癌细胞生物学行为的影响

目的：探讨钙结合蛋白S100A16在胃癌组织和细胞中的表达，及其对胃癌细胞SGC?7901增殖、迁移、侵袭能力的影响。方法：应用免疫组织化学染色法检测S100A16在胃癌和相应癌旁组织中的差异表达。应用Western blot检测正常胃上皮细胞GES?1以及胃癌细胞MGC?803和SGC?7901中S100A16的表达水平。将S100A16高表达质粒和干扰质粒分别转染至人胃癌细胞株SGC?7901中，用Western blot检测各组细胞中S100A16的表达。采用磺酰罗丹明B（sulforhodamine B，SRB）比色法和克隆形成实验检测细胞的增殖能力。采用划痕实验检测细胞迁移能力，采用Transwell实验检测细胞侵袭能力。结果：在胃癌组织及胃癌细胞中，S100A16的表达量明显高于相应癌旁组织及正常胃上皮细胞。SRB染色和克隆形成实验显示，过表达S100A16会增加SGC?7901细胞的增殖能力，敲低S100A16后增殖能力降低。划痕实验Transwell实验显示，过表达S100A16会增加SGC?7901细胞的迁移和侵袭能力，敲低S100A16则会降低。免疫共沉淀实验显示在胃癌细胞SGC?7901中，S100A16与YBX?1存在结合。结论：胃癌组织及细胞中S100A16的表达明显上调，S100A16在胃癌中的异常表达可能是由于与YBX?1的相互结合，进而促进胃癌细胞增殖、迁移、侵袭从而影响胃癌的发生发展。

Effects of calcium binding protein S100A16 on biological behavior of gastric cancer cells

Abstract] Objective: To investigate the expression of calcium binding protein S100A16 in gastric cancer tissues and cells and its effect on the proliferation, migration and invasion of gastric cancer cell SGC-7901. Methods: Immunohistochemical staining is used to detect the differential expression of S100A16 in 8 pairs of gastric cancer and corresponding adjacent tissues. S100A16 overexpression plasmid and empty plasmid, interference plasmid and empty vector are transfected into human gastric cancer cell line SGC-7901 respectively, and the expression of S100A16 in each group is detected by Western blot. SRB staining and colony formation assay are used to detect the proliferation of cells in each group. Scratch test is used to detect the migration ability of cells in each group. Transwell assay is used to detect the invasive ability of each group. Results: The expression of S100A16 in gastric cancer tissues and gastric cancer cells are significantly higher than that in corresponding paracancerous tissues and normal gastric epithelial cells. The expression levels of S100A16 in SGC-7901 and MGC-803 cells are 3.6 times and 1.4 times (p <0.05), respectively, of normal gastric epithelial cells GES-1. The S100A16 overexpression plasmid transfected SGC-7901 cells significantly up-regulated S100A16 expression, S100A16 expression level is 2.9 times of the control group (p <0.05); SGC-7901 cells transfected with S100A16 interference plasmid can significantly reduce S100A16 expression, S100A16 expression level is 61.6% of the control group (p<0.05). SRB staining and colony formation assays show that overexpression of S100A16 increases the proliferation of SGC-7901 cells and knocks down the proliferation of S100A16 cells. Scratch experiments show that overexpression of S100A16 increases SGC-7901 cell migration, while knockdown of S100A16 decreases. Transwell experiments show that overexpression of S100A16 increases the invasiveness of SGC-7901 cells and knockdown S100A16 decreases. Western blot analysis shows that the expression of YBX-1 in SGC-7901 cells transfected with S100A16 over-expression plasmid is significantly increased compares with the control group. Co-immunoprecipitation experiments show that S100A16 binds to YBX-1 in gastric cancer cell line SGC-7901. Conclusion:The expression of S100A16 in gastric cancer tissues and cells is significantly up-regulated. The abnormal expression of S100A16 in gastric cancer may be due to the interaction with YBX-1, thereby promoting the proliferation, migration, invasion of gastric cancer cells and affecting the occurrence and development of gastric cancer. Key words] Gastric cancer; Calcium binding protein S100A16; Proliferation; YBX-1