论文部分内容阅读
目的构建Foxp3基因的shRNA干扰表达载体,观察Foxp3基因表达对人胃癌细胞株BGC-823细胞增殖和凋亡的影响。方法根据人Foxp3序列设计并构建Foxp3基因的shRNA真核表达载体,利用脂质体LipofactamineTM2000转染胃癌细胞株,经G418筛选出稳定转染的阳性细胞,Real-time PCR检测在mRNA水平干扰质粒对Foxp3的抑制效应,MTT法、流式细胞术观察转染后细胞生长特性的变化。结果成功将含靶向Foxp3基因的短发夹双链RNA(shRNA)的重组质粒转染Foxp3表达阳性的胃癌细胞株BGC-823,转染shRNA-Foxp3重组质粒的BGC-823细胞Foxp3mRNA表达明显下降,与阴性对照组相比,差异有统计学意义(P<0.05)。转染干扰质粒抑制细胞增殖,促进细胞凋亡,与未转染组和阴性对照组相比,胃癌细胞生长周期出现阻滞在G0/G1期,差异有统计学意义(P<0.05)。结论干扰Foxp3的表达可显著抑制胃癌细胞的增殖和促进细胞凋亡。
Objective To construct shRNA interference expression vector of Foxp3 gene and observe the effect of Foxp3 gene expression on proliferation and apoptosis of human gastric cancer cell line BGC-823. Methods According to human Foxp3 sequence, a shRNA eukaryotic expression vector of Foxp3 gene was designed and constructed. LipofactamineTM2000 was used to transfect gastric cancer cell lines. Stably transfected cells were screened by G418. Real-time PCR was used to detect the expression of Foxp3 mRNA. The inhibitory effect of Foxp3, MTT assay, flow cytometry was observed after transfection cell growth characteristics. Results The recombinant plasmid containing short hairpin double stranded RNA targeting Foxp3 gene was successfully transfected into BGC-823 cell line expressing Foxp3. The expression of Foxp3 mRNA in BGC-823 cells transfected with shRNA-Foxp3 was significantly decreased , Compared with the negative control group, the difference was statistically significant (P <0.05). Transfection interference plasmid inhibited cell proliferation and promoted cell apoptosis. Compared with untransfected group and negative control group, the cell cycle of gastric cancer showed a block in G0 / G1 phase, the difference was statistically significant (P <0.05). Conclusion Interference of Foxp3 expression can significantly inhibit the proliferation of gastric cancer cells and promote apoptosis.