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目的探讨封闭人端粒酶RNA功能区的反义寡核苷酸(human telomerase RNA antisense oligonucleotide,hTR ASODN)对结直肠癌SW480细胞凋亡及凋亡诱导因子(apoptosis inducing factor,AIF)表达的影响。方法通过脂质体将hTR ASODN1次/d,连续3d转染入人结直肠癌SW480细胞,以未加ASODN的脂质体空白组及脂质体转染的正义组为对照。荧光显微镜观察转染情况,流式细胞仪测定转染后SW480细胞的凋亡率,RT-PCR法测定转染后AIF的表达情况。结果 hTR ASODN转染细胞后,倒置显微镜下观察各组细胞,细胞内有绿色荧光均匀分布表示转染良好。流式细胞仪测定转染24、48、72h后,各实验组凋亡率分别为:空白组(3.07±0.11)%、(3.71±0.63)%、(4.23±0.41)%,正义组(4.28±0.41)%、(4.49±0.56)%、(5.13±0.56)%,反义组(13.11±0.32)%、(22.04±0.21)%、(16.71±0.82)%,反义组分别与空白组、正义组比较,差异有统计学意义(P<0.05)。空白组、正义组、反义组光密度比值分别为(0.1527±0.0021)、(0.1580±0.0078)、(0.3683±0.0109),反义组分别与空白组、正义组比较,差异有统计学意义(P<0.05)。结论 hTR ASODN具有明显的促进凋亡、抗肿瘤作用。
Objective To investigate the effect of human telomerase RNA antisense oligonucleotide (hTR ASODN) blocking the telomerase RNA function on the apoptosis and the expression of apoptosis inducing factor (AIF) in colorectal cancer SW480 cells . Methods Human hTR ASODN1 was transfected into human colorectal cancer SW480 cells by liposomes once / d for 3 consecutive days. The ASODN-free liposome blank group and liposome-transfected positive control group were used as controls. Transfection conditions were observed by fluorescence microscopy. The apoptosis rate of SW480 cells was detected by flow cytometry. The expression of AIF was detected by RT-PCR. Results hTR ASODN transfected cells were observed under inverted microscope cells in each group, the cells were evenly distributed with green fluorescence that transfection was good. The apoptotic rate of each experimental group after transfection was 24,48,72 h by flow cytometry: blank group (3.07 ± 0.11)%, (3.71 ± 0.63)%, (4.23 ± 0.41)%, (4.49 ± 0.56)%, (5.13 ± 0.56)%, antisense group (13.11 ± 0.32)%, (22.04 ± 0.21)%, (16.71 ± 0.82)%, respectively. , The difference between the two groups was statistically significant (P <0.05). The optical density ratios of the blank group, the sense group and the antisense group were (0.1527 ± 0.0021) and (0.1580 ± 0.0078) and (0.3683 ± 0.0109) respectively. There was significant difference between the antisense group and the blank group and the sense group P <0.05). Conclusion hTR ASODN has a significant promotion of apoptosis, anti-tumor effect.