伴侣蛋白CCT6A参与结肠癌细胞迁移及侵袭的作用研究

Study on chaperonin CCT6A participating in the process of migration and invasion of colon cancer cell lines

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 explore the role of CCT6A in the migration and invasive abilities of human colon cancer cell lines.Methods Nine colon cancer cell lines were cultured.The cell line that expressed the highest level of CCT6A was selected by real-time PCR and Western-blotting.Three siRNA oligo segments targeting CCT6A were designed to transfect the selected cell lines;after transfection,the efficiency of siRNA oligo segment were detected by real-time PCR and Western-blotting.Furthermore,after being transfected with the most efficient siRNA oligo segment,the changes of migration and invasive capabilities of the selected cell line were evaluated by the Transwell test.Results Colon cancer cell line SW1116 was selected as having the highest expression of CCT6A on both the mRNA and the protein levels.The expression of CCT6A in SW1116 was reduced by all the three siRNA oligo segments to varying degrees;of them the No.2 siRNA oligo segment was the most efficient.In the migration test,the number of cells crossing through chambers in the CCT6A-siRNA group was（6±2）,which was lower than that of the negative-control（23±12） and the blank-control groups（21±11）;in the invasion test,the number of cells crossing through chambers in the PLK1-siRNA group was（9±2）,which was lower than that of the negative-control（40±12） and the blank-control groups（43±10）;the difference being significant statistically（P0.05）.Conclusions By knocking down chaperonin CCT6A,siRNA could significantly inhibit the migratory and invasive capabilities of the colon cancer cell lines;suggesting that CCT6A might play an important role in the infiltration and metastasis of colon cancer.