论文部分内容阅读
目的:探讨沉默磷脂酶D2(phospholipases D2,PLD2)基因的表达对人结肠癌细胞增殖和迁移能力的影响及其潜在的分子机制。方法:采用载有靶向PLD2的小干扰RNA(small interfering RNA,si RNA)腺病毒转染SW480和SW620细胞;q RTPCR和Western blot法检测转染后结肠癌细胞PLD2的m RNA和蛋白质表达的变化;平板克隆形成实验和Transwell小室分别检测各组细胞的增殖和细胞迁移能力的变化;采用Western blot法检测转染前后细胞中周期蛋白D1(Cyclin D1)和基质金属蛋白酶-2(metalloproteinase-2,MMP-2)蛋白的表达。结果:转染PLD2 si RNA后结肠癌细胞PLD2 m RNA和蛋白的表达量明显下降(均P<0.01),沉默PLD2表达后细胞的增殖和迁移能力明显受抑制,同时Cyclin D1和MMP-2蛋白的表达量也明显下调(均P=0.000)。结论:干扰PLD2表达引起的细胞增殖和迁移能力的下降可能与Cyclin D1和MMP-2表达下调密切相关,提示PLD2可能成为结肠癌治疗的新靶点之一。
Objective: To investigate the effects of phospholipase D2 (PLD2) gene on the proliferation and migration of human colon cancer cells and its potential molecular mechanisms. Methods: SW480 and SW620 cells were transfected with small interfering RNA (siRNA) targeting adenovirus targeting PLD2. The mRNA and protein expression of PLD2 in colon cancer cells were detected by qRTCR and Western blot The expression of Cyclin D1 and metalloproteinase-2 in transfected cells were detected by Western blot. The changes of cell proliferation and cell migration in each group were detected by plate clone formation assay and Transwell chamber. , MMP-2) protein expression. Results: The expression of PLD2 mRNA and protein in colon cancer cells was significantly decreased after transfected with PLD2 si RNA (all P <0.01), and the proliferation and migration ability of PLD2 cells was significantly inhibited after PLD2 silencing. Cyclin D1 and MMP-2 protein The expression level was also significantly down-regulated (P = 0.000). Conclusion: The decrease of cell proliferation and migration induced by interfering with PLD2 expression may be closely related to the down-regulation of Cyclin D1 and MMP-2, suggesting that PLD2 may be one of the new targets for the treatment of colon cancer.