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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。
The aim of this study was to investigate the effect of recombinant human S100A6 protein on the proliferation, apoptosis, migration and invasion of breast cancer cell line MCF-7. Recombinant human S100A6 protein (GST-hS100A6) was prepared by using prokaryotic expression. The size of the recombinant protein was 36 kDa by SDS-PAGE. Western blot showed that it could be recognized by S100A6 antibody. Compared with GST group, the D 492 values of GST-hS100A6 group increased by 29.1% and 84.6% (P <0.05) when treated with human breast cancer cell line MCF-7 at MTT for 48 h <0.05), suggesting that S100A6 promoted the proliferation of MCF-7 cells. The clonal formation assay showed that the clonogenic rate of GST-hS100A6 group was 38.7% higher than that of GST group (P <0.05), suggesting that S100A6 promoted the formation of MCF-7. Hoechst staining The results showed that the apoptotic rate of GST-hS100A6 group was 67.8% (P <0.05) lower than that of GST group at 24 h, and decreased 48.4% (P <0.05) at 48 h, suggesting that S100A6 inhibited MCF-7 Scratch test showed that the wound healing rate of GST-hS100A6 group was 2.2 times that of GST group (P <0.05) at 24 h, suggesting that S100A6 promoted the migration of MCF-7 cells; Transwell showed that GST-hS100A6 group at 24 h, the number of transmembrane cells increased by 88.1% (P <0.05) compared with GST group, suggesting that S100A6 promoted the invasion of MCF-7 cells. The above results show that S100A6 can promote human breast cancer and may become a new molecular marker for diagnosis and treatment of breast cancer.