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. 2012 Jun;18(6):BR237-46.
doi: 10.12659/msm.882900.

Polo-like kinase 1 is overexpressed in colorectal cancer and participates in the migration and invasion of colorectal cancer cells

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Polo-like kinase 1 is overexpressed in colorectal cancer and participates in the migration and invasion of colorectal cancer cells

Ding-pei Han et al. Med Sci Monit. 2012 Jun.

Abstract

Background: Polo-like kinase 1 (PLK1) is an important molecule in proliferation of many human cancers. The aim of study is to clarify the expression patterns and potential function of PLK1 in colorectal cancers.

Material/methods: Fifty-six colorectal cancers samples were collected and arranged onto a tissue array and the expression of PLK1 were detected by immunohistochemistry and correlated with clinico-pathological characteristics and expression of PCNA. Expression of PLK1 in 9 colorectal cancer cells lines was investigated by RT-PCR and Western blot, then SW1116 cells lines were treated with PLK1 siRNA and the efficiency was examined by Western blot. Transwell test was applied to detect the migration and invasion capability of cancer cells by counting the number of cells passing through the membranes. Cell proliferation and apoptosis were examined by Cell Counting Kit-8 (CCK-8) and Annexin-V Kit.

Results: PLK1 was positively expressed in 73.2% (41/56) of colorectal cancers tissues, but in only 3.6% (2/56) of normal tissues, and was associated with Duke's stage (P<0.01), tumor size (P<0.01), invasion extent (P<0.05) and lymphatic metastasis (P<0.01). The expression of PLK1 was correlated with expression of PCNA (R=0.553, P<0.01). PLK1 was inhibited in SW1116 cells by treating with PLK1 siRNA oligos, which resulted in a decreased number of cells passing through the membrane as compared with control groups (P<0.01) at 24 hours after transfection. Cell proliferation was inhibited from 48 hours after transfection, while cells apoptosis was induced from 72 hours after transfection.

Conclusions: PLK1 could be a progression marker for colorectal cancer patients and PLK1 depletion can inhibit migration and invasion capability of colorectal cancer cells SW1116, suggesting that PLK1 might be involved in metastasis and invasion of colorectal cancer. Therapeutic strategies targeting PLK1 may be a new approach to colorectal cancer.

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Figures

Figure 1
Figure 1
Expression of PLK1 and PCNA in colorectal tissues. (A) PLK1 negatively expressed in normal colorectal tissues; (B) PLK1 moderately expressed in colorectal cancer tissues; (C) PLK1 strongly expressed in colorectal cancer tissues; (D) PCNA negatively expressed in normal colorectal tissues; (E) PCNA moderately expressed in colorectal cancer tissues; (F) PCNA strongly expressed in colorectal cancer tissues. Original magnifications ×200.
Figure 2
Figure 2
Expression and inhibition of PLK1 in colorectal cancer cells lines. (A) PLK1 mRNA expression detected in 9 colorectal cancer cell lines by PCR; (B) PLK1 protein expression detected in 9 colorectal cancer cell lines by Western blotting; (C) PLK1 mRNA expression detected in siRNA-treated SW1116 by real-time PCR at 24 hours after transfection. Values are mean ±SD of the expression amount of PLK1 relative to GAPDH; (D) PLK1 protein expression detected in siRNA-treated SW1116 by Western blotting at 48 hours and 72 hours after transfection.
Figure 3
Figure 3
Effect of PLK1 depletion on migration and invasion of SW1116 cells. (A) 6.5 mm Transwell® with 8 μm pore Polycarbonate Membrane Insert were used for migration and invasion (with Matrigel) assay. The chambers were stained with methyl violet (×4 for migration, ×10 for invasion). (B) The migration and invasion cells were counted in 5 random fields of vision and the number of cells is expressed as mean ±SD for 3 replication experiments.
Figure 4
Figure 4
Effect of PLK1 depletion on cells proliferation and apoptosis. (A) CCK-8 was used for determining the role of PLK1 in regulating SW1116 proliferation at different moments. Values are the mean ±SD of absorbance at 450 nm for 6 replication experiments. (B,C) FCM was used to determine the role of PLK1 in regulating apoptosis of SW1116 at different moments. Annexin V and PI stained cells were detected by FCM. Values are the mean ±SD of apoptosis rate (including early and late apoptosis) for the 3 replication experiments.

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