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目的探讨survivin靶向SiRNA重组表达载体对人结直肠癌细胞生长的抑制作用和对5-氟尿嘧啶(5-FU)的化疗增敏作用。方法构建survivin靶向SiRNA重组表达载体并转染结肠癌细胞,通过RT-PCR和westernblot方法检测survivin的表达;采用MTT法和流式细胞仪检测SiRNA对不同浓度5-FU的化疗增敏作用。结果经酶切鉴定和测序结果证实survivin靶向SiRNA重组表达载体构建成功;其对结直肠癌细胞survivinmRNA和蛋白的表达抑制率分别为36.33%和44.65%,可以协同增强5-FU对结直肠癌细胞生长的抑制作用。结论survivin靶向SiRNA重组表达载体能有效抑制survivin基因表达,并能协同增强5-FU对结直肠癌细胞的杀伤作用和诱导凋亡作用。
OBJECTIVE: To investigate the inhibitory effect of survivin-targeting SiRNA recombinant vector on the growth of human colorectal cancer cells and the chemosensitivity to 5-fluorouracil (5-FU). Methods The survivin target siRNA recombinant vector was constructed and transfected into human colon cancer cells. The expression of survivin was detected by RT-PCR and western blot. The chemosensitization effect of SiRNA on 5-FU was detected by MTT assay and flow cytometry. Results Restriction endonuclease digestion and sequencing confirmed that survivin targeted siRNA recombinant expression vector was successfully constructed. The inhibitory rates of survivin mRNA and protein expression in colorectal cancer were 36.33% and 44.65%, respectively, which synergistically enhanced the inhibitory effect of 5-FU on colorectal cancer Inhibition of cell growth. Conclusion The survivin-targeting SiRNA recombinant expression vector can effectively inhibit the expression of survivin gene and can synergistically enhance the killing effect and induce the apoptosis of colorectal cancer cells by 5-FU.