FMNL2基因对大肠癌细胞SW620增殖、侵袭及转移的影响

发布时间：2018-08-04 18:32

【摘要】：目的探讨FMNL2基因对大肠癌细胞SW620增殖、侵袭及转移的影响。方法将大肠癌SW620细胞随机分为三组,其中稳定转染FMNL2-shRNA质粒的细胞作为shFMNL2组,稳定转染Pgenesil-1质粒的细胞作为对照组,单纯SW620细胞作为空白组。扫描电镜下观察三组细胞超微结构;Real-time PCR法检测FMNL2 mRNA表达,Western blotting法检测FMNL2蛋白表达,MTT法检测细胞增殖活性(光密度值),平板克隆形成试验检测细胞克隆数,Transwell小室及Boyden小室试验检测细胞运动及侵袭能力。结果透射电镜下,空白组细胞均具有癌细胞的一般特征;对照组细胞内无退变的细胞器,细胞质基质密度均匀,细胞表面可见许多微绒毛和伪足;shFMNL2组细胞质内出现大小不等空泡及自噬体,细胞质局部粗面消失,细胞膜及核膜基本完整,细胞表面绒毛大量减少,未发现伪足。shFMNL2组FMNL2 mRNA和蛋白相对表达量均低于对照组及空白组(P均0.05)。培养第1、2、3、4、5、6、7天时shFMNL2组细胞光密度值均低于对照组及空白组(P均0.05)。shFMNL2组细胞克隆数、Transwell小室及Boyden小室穿膜细胞数均较对照组及空白组减少(P均0.05)。结论抑制FMNL2基因表达导致大肠癌细胞增殖、侵袭、迁移能力下降;FMNL2基因可能是大肠癌侵袭、转移的相关基因之一,其可能作为大肠癌潜在的治疗靶点。
[Abstract]:Objective to investigate the effect of FMNL2 gene on the proliferation, invasion and metastasis of colorectal cancer cell line SW620. Methods Colorectal cancer SW620 cells were randomly divided into three groups: stable transfected FMNL2-shRNA plasmid cells as shFMNL2 group, stable transfected Pgenesil-1 plasmid cells as control group, simple SW620 cells as blank group. The expression of FMNL2 mRNA in three groups of cells was detected by Real-time PCR method. The expression of FMNL2 protein was detected by FMNL2 method. The cell proliferation activity (optical density) was detected by MTT assay. The number of cell clones in transwell chamber and Boyden were detected by plate clone formation test. Cell motility and invasiveness were measured by chamber test. Results under transmission electron microscope, all the cells in the blank group had the general characteristics of cancer cells, while in the control group, there was no degenerative organelle, and the density of the cytoplasmic matrix was uniform. Many microvilli and pseudopodium shFMNL2 cells were found to have unequal vacuoles and autophagy in the cytoplasm, the coarse surface of the cytoplasm disappeared, the cell membrane and nuclear membrane were basically intact, and the villi on the surface of the cells decreased significantly. The relative expression of FMNL2 mRNA and protein in the pseudopod. ShFMNL2 group was lower than that in the control group and the blank group (all P 0. 05). The cell optical density of shFMNL2 group was lower than that of control group and blank group (P 0.05). The number of transwell chamber and Boyden cell in shFMNL2 group were lower than those in control group and blank group (P 0.05). Conclusion inhibition of FMNL2 gene expression may lead to proliferation, invasion and migration of colorectal cancer cells. FMNL2 gene may be one of the genes associated with invasion and metastasis of colorectal cancer, which may be a potential therapeutic target for colorectal cancer.
【作者单位】：武警总医院;
【分类号】：R735.34

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