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目的观察Notch1基因沉默后人绒毛膜上皮癌细胞增殖及侵袭能力变化。方法查阅Gen Bank提供的Notch1序列(NM-001105721.1)设计引物,合成Notch1 siRNA。分离培养人绒毛膜上皮癌JEG-3细胞,分为A、B、C组。A组不做转染,B组转染control siRNA,C组转染Notch1 siRNA。分别于转染后12、24、36、48、60 h采用MTT法检测各组细胞增殖能力(OD值)。转染后48 h采用Transwell侵袭实验检测细胞侵袭能力。转染后72 h采用Western blotting法检测各组细胞中的MMP-2、MMP-9蛋白。结果转染后36、48、60 h C组OD值低于A、B组(P均<0.05)。A、B、C组穿膜细胞数分别为(109±6)、(118±7)、(59±4)个,C组与A、B相比,P均<0.05。A、B、C组细胞中MMP-2蛋白相对表达量分别为0.91±0.08、0.87±0.01、0.29±0.05,MMP-9蛋白相对表达量分别为0.85±0.04、0.79±0.03、0.35±0.05,C组MMP-2、MMP-9蛋白相对表达量与A、B组相比,P均<0.05。结论Notch1基因沉默后,人绒毛膜上皮癌细胞增殖受到抑制,侵袭能力减弱。
Objective To observe the changes of proliferation and invasion ability of human choriocarcinoma cells after Notch1 gene silencing. Methods Notch1 (GenBank accession number NM-001105721.1) was designed to amplify Notch1 siRNA. Human choriocarcinoma JEG-3 cells were isolated and cultured and divided into A, B and C groups. A group did not transfected, B group transfected control siRNA, C group transfected Notch1 siRNA. The cell proliferative ability (OD value) of each group was detected by MTT assay 12, 24, 36, 48 and 60 h after transfection respectively. Forty-eight hours after transfection, cell invasion was detected by Transwell invasion assay. The expression of MMP-2 and MMP-9 in each group was detected by Western blot 72 h after transfection. Results The OD value of C group at 36,48,60 h after transfection was lower than that of A and B groups (all P <0.05). The numbers of transmembrane cells in group A, B and C were (109 ± 6), (118 ± 7) and (59 ± 4), respectively, and those in group C were significantly lower than those in group A and B (P <0.05). The relative expression of MMP-2 protein in group A, B and C were 0.91 ± 0.08, 0.87 ± 0.01 and 0.29 ± 0.05 respectively, and the relative expression of MMP-9 protein were 0.85 ± 0.04, 0.79 ± 0.03 and 0.35 ± 0.05, The relative expression of MMP-2 and MMP-9 in group C was significantly lower than that in group A and B (P <0.05). Conclusion After silencing Notch1 gene, the proliferation of human choriocarcinoma cells is inhibited and the invasion ability is weakened.