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目的:研究白细胞介素-6(IL-6)反义核苷酸(ASODN)对人肝内胆管癌HCCC-9810细胞增殖和凋亡的影响。方法:通过脂质体LipofectamineTM2000介导IL-6ASODN转染人肝内胆管癌HCCC-9810细胞,细胞克隆率形成实验和MTT实验检测IL-6ASODN对胆管癌细胞增殖的影响,流式细胞仪检测IL-6ASODN对胆管癌细胞凋亡的影响,RT-PCR检测转染后IL-6mRNA表达水平的变化。结果:转染后,细胞克隆形成率反义组为(4.58±0.35)%,反义组显著低于各对照组(P<0.05);与空白组相比,反义组的细胞增殖抑制率为67.2%,反义组细胞增殖抑制率显著高于对照组(P<0.05);流式细胞仪检测反义组细胞的凋亡率为(10.25±0.13)%,显著高于对照组(P<0.05);反义组胆管癌细胞IL-6mRNA表达水平较空白组下降60.52%,能显著抑制IL-6mRNA的表达,较其余各组差异有显著性(P<0.05)。结论:IL-6反义寡核苷酸可以抑制胆管癌HCCC-9810细胞的增殖,并可以诱导其凋亡。
Objective: To investigate the effects of interleukin-6 antisense oligonucleotide (ASODN) on proliferation and apoptosis of human intrahepatic cholangiocarcinoma HCCC-9810 cells. METHODS: IL-6ASODN was transfected into human intrahepatic cholangiocarcinoma HCCC-9810 cells by LipofectamineTM2000, and the effect of IL-6ASODN on the proliferation of cholangiocarcinoma cells was detected by MTT assay. Flow cytometry was used to detect IL -6ASODN on apoptosis of cholangiocarcinoma cells. The expression of IL-6mRNA was detected by RT-PCR. Results: After transfection, the rate of cell clone formation was (4.58 ± 0.35)% in antisense group and significantly lower in antisense group than that in control group (P <0.05). Compared with the blank group, the cell proliferation inhibition rate (P <0.05). The apoptosis rate of antisense group was (10.25 ± 0.13)%, which was significantly higher than that of control group (P <0.05). Compared with the blank group, the expression of IL-6 mRNA in the antisense group of cholangiocarcinoma cells was decreased by 60.52%, and the expression of IL-6 mRNA was significantly inhibited compared with the other groups (P <0.05). Conclusion: IL-6 antisense oligonucleotide can inhibit the proliferation of HCCC-9810 cells and induce its apoptosis.