沉默Polo样激酶1基因的表达对结直肠癌细胞迁移和侵袭的影响

目的 研究PLK1在结直肠癌细胞迁移和侵袭过程中的作用.方法 常规培养9株结直肠癌细胞,采用PCR和Western-blot法筛选PLK1高表达细胞株.设计3种RNA干扰片段,转染高表达PLK1的结直肠癌细胞株,利用实时定量PCR和western-blot法验证并干扰效果.选择高干扰效果的干扰片段,通过Transwe1l实验来评估转染后结直肠癌细胞迁移和侵袭能力的变化.结果 SW1116细胞中PLK1 mRNA及蛋白都呈现高表达 3种干扰片段转染SW1116细胞后,PLK1表达量均有不同程度下降,以片段1干扰效果最为明显.在迁移实验中,PLK1干扰组穿膜细胞数为（44±14）个,低于阴性对照组[（242±40）个]和空白对照组[（240±38）个] 在侵袭实验中,PLK1干扰组穿膜细胞数为（62±3）个,低于阴性对照组[（207±12）个]和空白对照组[（211±15）个] 上述差异均有统计学意义（P＜0.01）.结论 干扰PLK1能显著抑制肿瘤细胞的迁移、侵袭能力,提示PLK1可能在结直肠癌浸润和转移中具有一定作用.

Influence of silencing Polo-like kinase 1 on migration and invasion of colorectal cancer cells

Objective To examine the role of Polo-like kinase 1 (PLK1) in the migration and invasiveness of human colorectal cancer cells. Methods Nine colorectal cancer cell lines were cultured. Cell lines with the highest level of PLK1 expression was selected by PCR and Western blot. Three siRNA oligos segments targeting PLK1 were designed and selected cell lines transfected. Successful transfection was confirmed using real-time PCR and Western blot. Changes in migration and invasiveness of the selected cell line were evaluated by Transwell test. Results Colorectal cancer cell line SW1116 was selected with the highest expression of PLK1 at both mRNA level and protein level. The expression of PLK1 in SW1116 was reduced by the three siRNA oligos segments to varying degrees, and the No.l siRNA oligos segment was the most efficient. In migration test, the number of cells crossing through chambers in PLKI-siRNA group was 44±14, which was lower than that in the negative control group (242±40) and in blank control group(240±38). In invasion test,the number of cells crossing through chambers in PLK1-siRNA group was 62±3, which was lower than that in negative control group (207±12) and in blank control group (211±15). These differences were statistically significant (P<0.01). Conclusion PLK1 silencing by siRNA may inhibit the migration and invasiveness of colorectal cancer cells, suggesting that PLK1 might play an important role in the infiltration and metastasis of colorectal cancer.