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目的:研究伴侣蛋白CCT6A在结肠癌细胞株迁移和侵袭过程中的作用。方法:培养9株结肠癌细胞,抽取总RNA和蛋白。采用实时定量PCR和Western印迹法筛选CCT6A高表达的细胞株。设计购买3种siRNA片段,转染高表达CCT6A的结肠癌细胞株,利用实时定量PCR和Western印迹法验证干扰效果。选择干扰效果最优的RNA干扰片段,通过Transwell实验来评估转染后结肠癌细胞迁移和侵袭能力的变化。结果:SW1116细胞株中的CCT6A在mRNA及蛋白水平都呈现高表达;3种干扰片段转染SW1116细胞后,CCT6A表达量均有不同程度下降,其中以片段2的干扰效果最优。在迁移实验中,CCT6A干扰组穿膜细胞数为(6±2)个,低于阴性对照组(23±12)个和空白对照组(21±11)个;在侵袭实验中,CCT6A干扰组穿膜细胞数为(9±2)个,低于阴性对照组(40±12)个和空白对照组(43±10)个;上述差异均有统计学意义(P<0.05)。结论:下调CCT6A的表达能显著抑制结肠肿瘤细胞的迁移、侵袭能力,提示CCT6A可能在结肠癌浸润和转移中起到一定作用。
Objective: To study the role of chaperone CCT6A in the migration and invasion of colon cancer cell lines. Methods: Nine colon cancer cells were cultured and total RNA and protein were extracted. Real-time quantitative PCR and Western blotting were used to screen the cell lines with high expression of CCT6A. Three siRNA fragments were purchased and transfected into CCT6A-overexpressing colon cancer cell lines. Real-time PCR and Western blotting were used to verify the interference effect. The RNA interference fragment with the best interference effect was selected and the changes of migration and invasion ability of colon cancer cells after transfection were evaluated by Transwell assay. Results: The expression of CCT6A in SW1116 cell line was highly expressed at mRNA and protein levels. The expression of CCT6A in SW1116 cells was decreased to varying degrees after transfection with three kinds of siRNAs. The interference effect of fragment 2 was the best. In the migration experiment, the number of transmembrane cells in the CCT6A interference group was (6 ± 2), which was lower than that in the negative control group (23 ± 12) and the blank control group (21 ± 11). In the invasion assay, the CCT6A interference group The number of transmembrane cells was (9 ± 2), which was lower than that in the negative control group (40 ± 12) and the blank control group (43 ± 10). The differences were statistically significant (P <0.05). Conclusion: The down-regulation of CCT6A can significantly inhibit the migration and invasion of colon tumor cells, suggesting that CCT6A may play a role in the invasion and metastasis of colon cancer.