Han Ding-Pei, Cui Jiang-Tao, Lu Ai-Guo, Chen Xue-Hua, Feng Bo, Zong Ya-Ping, Qu Shun, Cao Qi-Feng, Zheng Min-Hua
Department of General Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
Zhonghua Wei Chang Wai Ke Za Zhi. 2011 Jan;14(1):61-4.
To examine the role of Polo-like kinase 1(PLK1) in the migration and invasiveness of human colorectal cancer cells.
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 oligo 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.
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 oligo segments to varying degrees, and the No.1 siRNA oligo segment was the most efficient. In migration test, the number of cells crossing through chambers in PLK1-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).
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.
探讨Polo样激酶1（PLK1）在人结肠癌细胞迁移和侵袭中的作用。
培养9种结肠癌细胞系。通过PCR和蛋白质印迹法选择PLK1表达水平最高的细胞系。设计3条靶向PLK1的小干扰RNA（siRNA）寡核苷酸片段并转染所选细胞系。采用实时PCR和蛋白质印迹法确认转染成功。通过Transwell试验评估所选细胞系迁移和侵袭能力的变化。
结肠癌细胞系SW1116在mRNA水平和蛋白质水平上均具有最高的PLK1表达。3条siRNA寡核苷酸片段均不同程度降低了SW1116中PLK1的表达，其中1号siRNA寡核苷酸片段效果最佳。在迁移试验中，PLK1-siRNA组穿过小室的细胞数为44±14，低于阴性对照组（242±40）和空白对照组（240±38）。在侵袭试验中，PLK1-siRNA组穿过小室的细胞数为62±3，低于阴性对照组（207±12）和空白对照组（211±15）。这些差异具有统计学意义（P<0.01）。
siRNA沉默PLK1可能抑制结肠癌细胞的迁移和侵袭，提示PLK1可能在结肠癌的浸润和转移中起重要作用。
