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[目的]探讨c-Met siRNA对食管鳞癌细胞CE81T-4生物学特性的影响。[方法]利用si RNA技术,采用带有荧光的慢病毒包装的c-Met siRNA稳定转染CE81T-4,并设立转染慢病毒包装的c-Met siRNA的CE81T-4为实验组,转染空慢病毒作为阴性对照,未转染CE81T-4细胞作为空白对照。于荧光倒置显微镜下观察转染效果。采用qRT-PCR检测三组细胞中c-Met m RNA的表达量,Western blot检测c-Met蛋白表达。利用MTT、划痕实验、Transwell及流式细胞术分别检测细胞的增殖、迁移、侵袭、周期及凋亡。[结果]转染c-Met siRNA后,荧光倒置显微镜下可见广泛绿色荧光表达;实验组c-Met mRNA的表达量(0.1758±0.0150)显著低于阴性对照组(0.3384±0.0346)和空白对照组(0.3759±0.0252)(P<0.05)。实验组c-Met蛋白表达显影较空白对照组及阴性对照组低;实验组细胞的增殖能力显著降低,与阴性对照组和空白对照组比较差异有统计学意义(P<0.05)。Transwell体外侵袭实验提示实验组的侵袭迁移个数为135±11.79,显著低于空白对照组(186±14.98)及阴性对照组(178±12.53)(P<0.05)。实验组细胞凋亡率(17.87%±1.60%)高于阴性对照组(4.93%±2.49%)及空白对照组(4.37%±1.60%)(P<0.05)。实验组G0/G1期细胞比例(48.57%±4.91%)较阴性对照组(38.13%±3.23%)和空白对照组(37.07%±2.32%)显著增多(P<0.05)。[结论]c-Met siRNA可增加食管鳞癌细胞的凋亡率,阻滞细胞周期在G0/G1期,降低细胞增殖、迁移及侵袭能力。
[Objective] To investigate the effect of c-Met siRNA on the biological characteristics of esophageal squamous cell carcinoma CE81T-4 cells. [Methods] The CE81T-4 cells were stably transfected with the lentivirus-encapsulated c-Met siRNA by si RNA technology and the CE81T-4 cells transfected with the lentivirus-packaged c-Met siRNA were transfected Empty lentivirus as a negative control, not transfected CE81T-4 cells as a blank control. Transfection efficiency was observed under a fluorescence inverted microscope. QRT-PCR was used to detect the expression of c-Met m RNA in three groups of cells, Western blot was used to detect the expression of c-Met protein. Cell proliferation, migration, invasion, cycle and apoptosis were detected by MTT assay, scratch assay, Transwell assay and flow cytometry. [Results] The expression of c-Met mRNA in the experimental group (0.1758 ± 0.0150) was significantly lower than that in the negative control group (0.3384 ± 0.0346) and the blank control group (0.3759 ± 0.0252) (P <0.05). The expression of c-Met protein in experimental group was lower than that in blank control group and negative control group. The proliferation ability of experimental group was significantly lower than that in negative control group and blank control group (P <0.05). Transwell in vitro invasion assay showed that the number of invasion and migration was 135 ± 11.79 in the experimental group, which was significantly lower than that in the blank control group (186 ± 14.98) and the negative control group (178 ± 12.53) (P <0.05). The apoptotic rate of experimental group (17.87% ± 1.60%) was higher than that of negative control group (4.93% ± 2.49%) and blank control group (4.37% ± 1.60%) (P <0.05). Compared with the negative control group (38.13% ± 3.23%) and the blank control group (37.07% ± 2.32%), the percentage of cells in the experimental group at G0 / G1 phase (48.57% ± 4.91%) was significantly increased (P <0.05). [Conclusion] c-Met siRNA can increase the apoptosis rate of esophageal squamous cell carcinoma cells, arrest the cell cycle in G0 / G1 phase and decrease the cell proliferation, migration and invasion ability.