沉默PLD2基因对结肠癌细胞增殖和迁移能力的影响  

Effect of PLD2 gene silencing on proliferation and migration of colon cancer cells

作　　者：杜昆利 刘茂希[1] 张寿儒[1] 吴星烨[1] 曾丽[2] 傅仲学[1] 

机构地区：[1]重庆医科大学附属第一医院胃肠外科,重庆400016 [2]重庆医科大学附属第一医院中西医结合科,重庆400016

出　　处：《重庆医科大学学报》2016年第3期217-222,共6页Journal of Chongqing Medical University

基　　金：国家自然科学基金青年科学基金资助项目(编号:81201916);国家自然科学基金面上资助项目(编号:81172295)

摘　　要：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 effect of silencing the PLD2（phospholipases D2） gene expression on cell proliferation and migration of colon cancer cell lines and to explore the underline molecular mechanisms. Methods：The small interfering RNA（siRNA） targeted PLD2 was transfected into SW480 and SW620 cells by adenovirus. Expression levels of PLD2 mRNA and protein were mea- sured by qRT-PCR and Western blot,respectively. Plate clone formation assay and transwell chamber were used to detect the prolifera- tion and migration ability of colon cancer cells. Finally,the expressions of Cyclin D1 and metalloproteinase-2（MMP-2） protein were measured by Western blot in both cell lines. Results：PLD2 siRNA greatly inhibited the expression of PLD2 in colon cancer cells at both mRNA and protein levels（all P〈0.01）. Down-regulated the PLD2 expression significantly suppressed the proliferation and mi- gration ability of SW480 and SW620 cells and also inhibited the expression of Cyclin D1 and MMP-2 proteins in both cell lines（all P=0.000）. Conclusion： Down-regulated PLD2 gene expression can clearly inhibit the proliferation and migration of colon can- cer cells, which may closely associate with the decreased expression of Cyclin D1 and MMP-2,indicating a way of gene therapy for colon cancer.

关 键 词：磷脂酶D2 细胞增殖 细胞迁移 结肠癌 小干扰RNA 

分 类 号：R735.3[医药卫生—肿瘤]