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目的:探讨磷脂酶Cε1(phospholipase C epsilon-1,PLCE1)基因过表达对结肠癌细胞迁移、细胞周期及凋亡等生物学特性的影响。方法:采用脂质体转染的方法建立PLCE1过表达的SW620细胞株,实验分为3组:亲本细胞组(未转染基因),对照组[转染含绿色荧光蛋白(greenfluorescent protein,GFP)的空质粒],实验组(转染质粒pcDNA-DEST53-PLCE1)。分别采用实时荧光定量-PCR(real-time fluorogenic quantitative-PCR,RFQPCR)和蛋白质印迹法检测PLCE1 mRNA和蛋白在SW620细胞中的表达;Transwell小室法检测PLCE1过表达对SW620细胞迁移能力的影响,FCM法检测对细胞周期及凋亡率的影响,DNA Ladder法检测对细胞凋亡的影响。结果:PLCE1过表达可抑制结肠癌SW620细胞的迁移能力,实验组迁移细胞数为(8.80±1.72)个,而亲本组和对照组分别为(32.6±2.42)和(32.2±3.25)个,3组间差异有统计学意义(P<0.01)。PLCE1过表达可导致结肠癌细胞周期G1期延长,并出现凋亡。结论:PLCE1过表达可抑制结肠癌细胞的迁移能力,并引起细胞凋亡。PLCE1基因过表达使SW620结肠癌细胞恶性程度降低,该基因可能是新的结肠癌相关抑癌基因。
Objective: To investigate the effect of phospholipase C epsilon-1 (PLCE1) gene overexpression on the biological characteristics of colon cancer cell migration, cell cycle and apoptosis. METHODS: PLCE1 overexpressing SW620 cell line was established by liposome transfection. The experiment was divided into 3 groups: parental cell group (untransfected gene) and control group [transfected with green fluorescent protein (GFP) Empty plasmid], experimental group (transfected plasmid pcDNA-DEST53-PLCE1). The expression of PLCE1 mRNA and protein in SW620 cells was detected by real-time fluorogenic quantitative PCR (RFQPCR) and Western blotting, and the effect of PLCE1 overexpression on the migration of SW620 cells was detected by Transwell chamber method. The effect of the assay on cell cycle and apoptosis rate was examined by DNA Ladder assay. RESULTS: Overexpression of PLCE1 inhibited the migration of colon cancer SW620 cells. The number of migrating cells in the experimental group was (8.80±1.72), compared with (32.6±2.42) and (32.2±3.25) in the parental and control groups, respectively. The difference between the three groups was statistically significant (P<0.01). Overexpression of PLCE1 can lead to a prolonged G1 phase of colon cancer cells and apoptosis. Conclusion: Overexpression of PLCE1 can inhibit the migration of colon cancer cells and induce apoptosis. Overexpression of PLCE1 gene reduced the malignant degree of SW620 colorectal cancer cells, which may be a novel colon cancer-related anti-oncogene.