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目的研究cFLIP反义寡核苷酸(cFLIP-asODN)对人结肠癌细胞株SW620细胞增殖的影响。方法利用脂质体将cFLIP-asODN转染入SW620细胞,并设空载体+cFLIP-asODN转染组、脂质体+无意义ODN转染组和空白对照组作为对照。荧光倒置显微镜检测复合物对细胞的转染效率;荧光实时定量RT-PCR和Western blot检测转染后细胞中cFLIP mRNA和蛋白的表达水平的变化;MTT法分析复合物对SW620细胞增殖的影响;流式细胞术分析转染后细胞凋亡情况。结果与空载体+cFLIP-asODN转染组、脂质体+无意义ODN转染组和空白对照组相比,脂质体递送cFLIP-asODN进入细胞后,SW620细胞的增殖受到明显抑制(P<0.05);转染cFLIP-asODN后,SW620细胞中的cFLIP mRNA和蛋白的表达水平降低(P<0.05);且细胞凋亡率(29.5±2.4)%也明显高于空载体+cFLIP-asODN转染组(5.8±0.7)%、脂质体+无意义ODN转染组(5.5±0.8)%和空白对照组(5.2±0.5)%(P<0.05)。结论通过在体外转染cFLIP-asODN可有效抑制结肠癌细胞株SW620的增殖。
Objective To investigate the effect of cFLIP antisense oligonucleotide (cFLIP-asODN) on the proliferation of human colon carcinoma cell line SW620. Methods The cFLIP-asODN was transfected into SW620 cells by lipofectamine. The empty vector + cFLIP-asODN transfection group, liposome + non-sense ODN transfection group and blank control group were used as control. Fluorescence inverted microscope was used to detect the transfection efficiency of the complex on the cells. Fluorescence real-time quantitative RT-PCR and Western blot were used to detect the expression of cFLIP mRNA and protein in the transfected cells. MTT assay was used to analyze the effect of the complex on the proliferation of SW620 cells. Flow cytometry analysis of apoptosis after transfection. Results Compared with the empty vector + cFLIP-asODN transfection group, the liposome + non-sense ODN transfection group and the blank control group, the proliferation of SW620 cells was significantly inhibited after cFLIP-asODN was delivered into the cells (P < 0.05). The expression of cFLIP mRNA and protein in SW620 cells was decreased after transfected with cFLIP-asODN (P <0.05), and the apoptosis rate (29.5 ± 2.4)% was also significantly higher than that of empty vector + cFLIP-asODN transfected (5.8 ± 0.7)% in the control group, 5.5 ± 0.8% in the liposome + non-sense ODN transfected group and 5.2 ± 0.5% (P <0.05) in the blank control group. Conclusion The proliferation of colon cancer cell line SW620 can be effectively inhibited by transfecting cFLIP-asODN in vitro.