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目的 DNA结合分化抑制因子-1(inhibitor-1of DNA binding/differentiation-1,Id-1)对结肠肿瘤细胞侵袭行为影响的研究较少,本研究探讨siRNA转染下调Id-1表达对人结肠癌SW480细胞侵袭和转移行为的影响。方法将合成Id-1干扰序列(siRNA)转染至SW480细胞作为实验组(抑制组),并以Id-1表达干扰空白组、空载体组作为对照,应用反转录聚合酶链反应(RT-PCR)和蛋白质印迹法检测各组细胞中Id-1和基质金属蛋白酶-9(matrix metalloprotein,MMP-9)的表达情况,四甲基偶氮唑盐(MTT)法检测细胞的增殖能力;划痕损伤实验、Transwell小室和Matrigel侵袭模型评价Id-1对细胞侵袭和转移能力的影响。结果抑制组Id-1mRNA表达水平为(0.14±0.02),与空白组(1.27±0.03)和空载体组(1.25±0.06)比较明显降低,差异有统计学意义,P<0.001;抑制组Id-1蛋白表达水平为0.25±0.02,与空白组(1.18±0.03)和空载体组(1.16±0.04)比较明显降低,差异有统计学意义,P<0.001;抑制组MMP-9mRNA表达水平为0.19±0.02,与空白组(1.33±0.04)和空载体组(1.38±0.03)比较明显降低,差异有统计学意义,P<0.001;抑制组MMP-9蛋白表达水平为0.12±0.02,与空白组(0.89±0.04)和空载体组(0.91±0.02)比较明显降低,差异有统计学意义,P<0.001;生长曲线表明,从第3天起抑制组细胞增殖明显减弱,与空白组和空载体组差异明显;划痕损伤实验结果显示,空白组和空载体组向划痕处的迁移速度明显快于抑制组;迁移试验显示,抑制组迁移细胞数为75±12,明显少于空白组(201±12)和空载体组(206±15);抑制组侵袭细胞数为51±10,明显少于空白组(121±17)和空载体组(126±14),差异均有统计学意义,P<0.001。结论 Id-1促进SW480细胞的侵袭和转移,其机制可能与诱导MMP-9的表达有关。
Objective To investigate the effect of Id-1 on the invasiveness of colon tumor cells. The aim of this study was to investigate the effects of siRNA-transfected Id-1 on colon cancer Influence of invasion and metastasis of SW480 cells. Methods The transfected Id-1 siRNA was transfected into SW480 cells as experimental group (suppression group). Id-1 interference group and empty vector group were used as control. Reverse transcriptase-polymerase chain reaction The expression of Id-1 and matrix metalloprotein-9 (MMP-9) in each group were detected by Western blotting and Western blotting. The proliferation of cells was detected by MTT assay. Scratch-injury experiments, Transwell chamber and Matrigel invasion models were used to evaluate the effect of Id-1 on cell invasion and metastasis. Results Compared with the blank control group (1.27 ± 0.03) and empty vector control group (1.25 ± 0.06), the expression of Id-1 mRNA in the inhibition group was significantly lower (P <0.001) 1 protein expression was 0.25 ± 0.02, which was significantly lower than that of the blank group (1.18 ± 0.03) and the empty vector group (1.16 ± 0.04), P <0.001; the expression level of MMP-9 mRNA in the inhibition group was 0.19 ± (P <0.001). Compared with the blank group (1.33 ± 0.04) and the empty vector group (1.38 ± 0.03), the difference was statistically significant 0.89 ± 0.04) and empty vector group (0.91 ± 0.02), the difference was statistically significant (P <0.001). The growth curve showed that the proliferation of the inhibition group was obviously weakened from day 3, The results of scratch test showed that the migrating speed of the blank group and empty vector group to scratches was significantly faster than that of the inhibition group. The migration test showed that the number of migrating cells in the inhibition group was 75 ± 12, significantly less than that of the blank group ± 12) and empty vector group (206 ± 15). The number of invasive cells in the inhibition group was 51 ± 10, which was significantly lower than that in the blank group (121 ± 17) and the empty vector group 14), the difference was significant, P <0.001. Conclusion Id-1 can promote the invasion and metastasis of SW480 cells, which may be related to the induction of MMP-9 expression.