Tian Hua, Chen Yang, Zhao Jiangang, Liu Daren, Liang Gang, Gong Weihua, Chen Li, Wu Yulian
Department of General Surgery, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2017 Jul 25;46(4):341-348. doi: 10.3785/j.issn.1008-9292.2017.08.01.
To investigate the effects of siRNAs targeting CD97 immune epitopes on proliferation, infiltration, apoptosis and cell cycle of breast cancer cells.
siRNA sequences targeting CD97 and CD97 immune epitopes were designed according to Gene Bank NM_001025160.2 with smart siCatch siRNA design software. CD97siRNAs were transfected into MDA-MB231 cells in which CD97 was highly expressed. Highest sensitive CD97 and CD97 siRNA were screened by Western blotting. Inverted microscope was used to observe the growth of CD97siRNAs-transfected MDA-MB231 cells; the proliferation activity of MDA-MB231 cells was detected by MTT method; the wound healing assay and Transwell migration test were performed to examine the migration and infiltration ability of CD97 and CD97 siRNA-transfected MDA-MB231 cells; the effects of CD97 siRNA and CD97 siRNA on cell apoptosis and cell cycle of MDA-MB231 cells were detected by TUNEL and flow cytometry.
The growth and proliferation activity of CD97siRNAs-transfected MDA-MB231 cells were significantly lower than those in the control groups, and such differences were more significant in CD97 siRNA-transfected group (all <0.05); scratch test showed that the wound healing rate was lower in CD97siRNAs-transfected groups, especially in CD97 siRNA-transfected group (all <0.05); Transwell migration showed that the number of MDA-MB231 cells crossing through chambers were less in CD97siRNAs-transfected groups, especially in CD97 siRNA-transfected group (all <0.05); no significant difference in cell apoptosis was observed between CD97siRNAs-transfected groups and control groups; cell cycle detection showed that CD97siRNAs-transfected groups had less cells in G/G phase and more cells in S phase compared with the control groups, and such effect on cell cycle was more marked in CD97 siRNA-transfected group (all <0.05).
CD97 plays an important role in the cell growth, proliferation, migration and invasion of breast cancer MDA-MB231 cells, and compared with CD97, CD97 may have more effective inhibitory effects on cellular malignant behaviors.
研究靶向CD97免疫表位的小干扰RNA（siRNAs）对乳腺癌细胞增殖、浸润、凋亡及细胞周期的影响。
根据基因库NM_001025160.2，采用智能siCatch siRNA设计软件设计靶向CD97及CD97免疫表位的siRNA序列。将CD97 siRNAs转染至CD97高表达的MDA-MB231细胞中。通过蛋白质免疫印迹法筛选出敏感性最高的CD97及CD97 siRNA。利用倒置显微镜观察转染CD97 siRNAs的MDA-MB231细胞的生长情况；采用MTT法检测MDA-MB231细胞的增殖活性；通过划痕实验和Transwell迁移实验检测转染CD97及CD97 siRNA的MDA-MB231细胞的迁移和浸润能力；采用TUNEL法和流式细胞术检测CD97 siRNA及CD97 siRNA对MDA-MB231细胞凋亡和细胞周期的影响。
转染CD97 siRNAs的MDA-MB231细胞的生长和增殖活性明显低于对照组，且在转染CD97 siRNA的组中差异更显著（均<0.05）；划痕实验显示，转染CD97 siRNAs的组伤口愈合率较低，尤其是转染CD97 siRNA的组（均<0.05）；Transwell迁移实验显示，转染CD97 siRNAs的组中穿过小室的MDA-MB231细胞数量较少，尤其是转染CD97 siRNA的组（均<0.05）；转染CD97 siRNAs的组与对照组之间细胞凋亡无明显差异；细胞周期检测显示，与对照组相比，转染CD97 siRNAs的组G0/G1期细胞较少，S期细胞较多，且转染CD97 siRNA的组对细胞周期的影响更明显（均<0.05）。
CD97在乳腺癌MDA-MB231细胞的生长、增殖、迁移和侵袭中起重要作用，与CD97相比，CD97对细胞恶性行为可能具有更有效的抑制作用。
