重组人S100A6促进人乳腺癌细胞MCF-7的增殖和迁移侵袭并抑制其凋亡  

Recombinant Human S100A6 Promotes the Proliferation,Migration and Invasion but Inhibits the Apoptosis of Human Breast Cancer Cell Line MCF-7

作　　者：游莉[1] 徐兰兰[1] 郭元元[1] 孙双双[1] 邹正渝[1] 黎玉叶[1] 罗进勇[1] 何通川[1] 周兰[1] 

机构地区：[1]重庆医科大学医学检验系临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国细胞生物学学报》2011年第7期746-751,共6页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(No.30772548)资助项目~~

摘　　要：该研究旨在探讨重组人S100A6蛋白对乳腺癌细胞株MCF-7的增殖、凋亡、迁移及侵袭能力的影响。利用原核表达制备重组人S100A6蛋白(GST-hS100A6),SDS-PAGE显示其大小为36 kDa,Western blot显示其可以被S100A6抗体特异识别,BCA法测定1 L菌液共收获约16.7 mg蛋白;将其作用于人乳腺癌细胞MCF-7,MTT显示细胞培养48 h时,浓度为100μg/mL和300μg/mL的GST-hS100A6组的D_(492)值较GST组增加29.1%和84.6%(P<0.05),提示S100A6促进MCF-7细胞增殖;平板克隆形成实验显示GST-hS100A6组的克隆形成率较GST组高38.7%(P<0.05),提示S100A6促进MCF-7的克隆形成;Hoechst染色显示GST-hS100A6组在24 h时细胞凋亡率较GST组减少67.8%(P<0.05),48 h时细胞凋亡率较GST组减少58.4%(P<0.05),提示S100A6抑制MCF-7细胞凋亡;划痕实验显示在24 h时GST-hS100A6组的划痕愈合率为GST组的2.2倍(P<0.05),提示S100A6促进MCF-7细胞迁移;Transwell显示GST-hS100A6组在24 h时穿膜细胞数较GST组增加88.1%(P<0.05),提示S100A6促进MCF-7细胞侵袭。以上结果显示S100A6对人乳腺癌具有一定的促进作用,有可能成为乳腺癌分子诊断的标志物和治疗的新靶标。To aim at effects of human S 100A6 on proliferation, apoptosis, migration and invasion of human breast cancer cell line MCF-7, recombinant protein GST-hS 100A6 was purified from bacteria BL21 that identi- fied as 36 kDa by SDS-PAGE and recognized by S100A6 antibody with Western blot. MCF-7 were treated by GST- hS 100A6 with different concentrations while GST as a control group. MTT was used to detect the cell proliferation. Results showed that after 48 h, the D492 value of 100 μg/mL and 300 p.g/mL group of GST-hS100A6 increased by 29.1% and 84.6% compared with GST group, respectively （P〈0.05）. Colony-forming assay showed that the abil- ity of colony formation of GST-hS100A6 increased by 38.7% compared with GST group （P〈0.05）. Hoechst stain- ing showed that cell apoptosis rate of GST-hS100A6 group decreased by 67.8% compared with GST group at 24 h （P〈0.05） and decreased by 58.4% at 48 h （P〈0.05）. Results of wound healing assay indicated that the healing rate of GST-hS100A6 group was 2.2 times of GST group （P〈0.05） at 24 h. Transwell invasion assay showed that the trans- membrane cell number of GST-hS 100A6 group increased by 88.1% （P〈0.05） compared with GST group at 24 h. To sum up, S 100A6 could promote cell proliferation, colony formation, migration, and invasion, but inhibit cell apop- tosis on human breast cancer cell line MCF-7, indicates that S100A6 has a promoting effect on human breast cancer which would be a new molecular target for treatment of human breast cancer.

关 键 词：S100A6 乳腺癌 增殖 凋亡 迁移 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]