Expression and Significance of PLK1 and STK15 Gene, and Effect of Its Specific Inhibitor on Proliferation in Colon Cancer Cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 GAOJun-yong ¹ ,  YUShao-hong ² , ZHAOYong-heng ² , WANGWen-xue ³ , ZHOUQi ¹ , LIMei-zhang ³

1. Department of Oncology, Fuling Center Hospital of Chongqing, Chongqing 400800, China;
2. Department of General Surgery, Kunming 1 st People's Hospital, Kunming 650011, Yunnan Province, China;
3. School of Life Sciences, Yunnan University, Kunming 650000, Yunnan Province, China;

Corresponding author：

YUShao-hong, Email: ysh197458@sina.com

Keywords：

Polo-like kinase 1; Serine/threonine kinase 15; Colon cancer; SBE13; VX-680

DOI：

10.7507/1007-9424.20140168

Video：

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Objective To explore the expressions of polo-like kinase 1(PLK1) and serine/threonine kinase 15 (STK15) mRNA and protein in colon cancer cells, and to explore the inhibitive effect of SBE13 and VX-680 for PLK1 protein and STK15 protein. Methods One kind of cervical cancer cells(Hela cells) and 3 kinds of colon cancer cells (HCT-116 cells, HT-29 cells, and CACO-2 cells) were selected for experiment. Expression levels of PLK1 mRNA, STK15 mRNA and its protein of 4 kinds of cells were detected by reverse transcription polymerase chain reaction(RT-PCR) and Western blot method respectively. Inhibitive effect of SBE13 and VX-680 were evaluated in vitro by methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay in 4 kinds of cells, which divided into 5 groups, receiving Dulbecco's modification of Eagle's medium(DMEM), dimethylsulfoxide(DMSO), SBE13, VX-680, and SBE13+VX-680 respectively. Results Compared with Hela cells, expression levels of PLK1 mRNA, STK15 mRNA and its protein in HCT-116 cells,HT-29 cells, and CACO-2 cells were higher(P<0.05). ① Hela cells:Compared with DMEM group, the proliferative activity were not inhibited in SBE13 group, VX-680 group, and SBE13+VX-680 group(P>0.05). ② HCT-116 cells and HT-29 cells:Compared with DMEM group, the proliferative activity were inhibited in VX-680 group and SBE13+VX-680 group(P<0.05), but was not inhibited in SBE13 group(P>0.05). ③ CACO-2 cell:Compared with DMEM group, the proliferative activity were inhibited in SBE13 group, VX-680 group, and SBE13+VX-680 group(P<0.05). Conclusions Expression levels of PLK1 mRNA, STK15 mRNA and its protein increase in HCT-116, HT-29, and CACO-2 cells compared with Hela cells. SBE13 and VX-680 can inhibit PLK1 and STK15 protein partly in colon cancer cell lines.

Citation： GAOJun-yong, YUShao-hong, ZHAOYong-heng, WANGWen-xue, ZHOUQi, LIMei-zhang. Expression and Significance of PLK1 and STK15 Gene, and Effect of Its Specific Inhibitor on Proliferation in Colon Cancer Cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2014, 21(6): 702-706. doi: 10.7507/1007-9424.20140168 Copy

1.	陈金明,李伟华,王家顿,等.结肠癌细胞凋亡与增殖的平衡及其与预后的关系[J].癌症, 2005, 24(5):554-558.
2.	Hörlin A, Krause J, Radü U, et al.Tumor proliferation, MDR phenotype and tumor associated t-lymphocytes in non-Hodgkin's lymphomas[J].Verh Dtsch Ges Pathol, 1992, 76:248-252.
3.	Li GC, Zhang Z, Ma XJ, et al.Are biomarkers correlated with recurrence patterns in patients with resectable gastric adenocarcinoma[J].Mol Biol Rep, 2012, 39(1):399-405.
4.	Feng W, Malpica A, Skaland I, et al.Can proliferation biomarkers reliably predict recurrence in World Health Organization 2003 defined endometrial stromal sarcoma, low grade?[J].PLoS One, 2013, 8(10):e75899.
5.	Hellman K, Johansson H, Andersson S, et al.Prognostic significance of cell cycle-and invasion-related molecular markers and genomic instability in primary carcinoma of the vagina[J].Int J Gynecol Cancer, 2013, 23(1):41-51.
6.	Toyoshima-Morimoto F, Taniguchi E, Nishida E.Plk1 promotes nuclear translocation of human Cdc25C during prophase[J].EMBO Rep, 2002, 3(4):341-348.
7.	兰斌,刘炳亚,朱正纲.PLK1在细胞周期生物学的研究进展[J].医学分子生物学杂志, 2005, 2(1):65-67.
8.	Katayama H, Sasai K, Kawai H, et al.Phosphorylation by aurora kinase A induces Mdm2-mediated destabilization and inhibition of p53[J].Nat Genet, 2004, 36(1):55-62.
9.	Tsai MY, Wiese C, Cao K, et al.A ran signalling pathway mediated by the mitotic kinase Aurora a in spindle assembly[J].Nat Cell Biol, 2003, 5(3):242-248.
10.	Sen S, Zhou H, Zhang RD, et al.Amplification/overexpression of a mitotic kinase gene in human bladder cancer[J].J Natl Cancer Inst, 2002, 94(17):1320-1329.
11.	Zhou H, Kuang J, Zhong L, et al.Tumour amplified kinase STK15/BTAK induces centrosome amplification, aneuploidy and transformation[J].Nat Genet, 1998, 20(2):189-193.
12.	Ouchi M, Fujiuchi N, Sasai K, et al.BRCA1 phosphorylation by Aurora-A in the regulation of G2 to M transition[J].J Biol Chem, 2004, 279(19):19643-19648.
13.	兰斌,陈雪华,刘炳亚,等.STK15基因沉默导致胃癌细胞分裂停滞[J].中华病理学杂志, 2006, 35(2):106-109.
14.	Hannak E, Kirkham M, Hyman AA, et al.Aurora-A kinase is required for centrosome maturation in caenorhabditis elegans[J].J Cell Biol, 2001, 155(7):1109-1116.
15.	Takahashi T, Sano B, Nagata T, et al.Polo-like kinase 1(PLK1) is overexpressed in primary colorectal cancers[J].Cancer Sci, 2003, 94(2):148-152.
16.	Spänkuch-Schmitt B, Bereiter-Hahn J, Kaufmann M, et al.Effect of RNA silencing of polo-like kinase-1(PLK1) on apoptosis and spindle formation in human cancer cells[J].J Natl Cancer Inst, 2002, 94(24):1863-1877.
17.	Weichert W, Kristiansen G, Schmidt M, et al.Polo-like kinase 1 expression is a prognostic factor in human colon cancer[J].World J Gastroenterol, 2005, 11(36):5644-5650.
18.	欧海玲,覃宇周,王进声.丝氨酸/苏氨酸激酶15在早期乳腺癌组织中的表达及与人类表皮生长因子受体2、雌激素受体的关系[J].实用医学杂志, 2010, 26(19):3498-3500.
19.	宋波,郝钢跃,马蔚然,等.STK15基因多态性与膀胱癌发病风险的关系[J].现代泌尿外科杂志, 2010, 15(4):277-279.
20.	Golsteyn RM, Mundt KE, Fry AM, et al.Cell cycle regulation of the activity and subcellular localization of Plk1, a human protein kinase implicated in mitotic spindle function[J].J Cell Biol, 1995, 129(6):1617-1628.
21.	Lee KS, Yuan YL, Kuriyama R, et al.Plk is an M-phase-specific protein kinase and interacts with a kinesin-like protein, CHO1/MKLP-1[J].Mol Cell Biol, 1995, 15(12):7143-7151.
22.	Feng Y, Longo DL, Ferris DK.Polo-like kinase interacts with proteasomes and regulates their activity[J].Cell Growth Differ, 2001, 12(1):29-37.
23.	Roshak AK, Capper EA, Imburgia C, et al.The human pololike kinase, PLK, regulates cdc2/cyclin B through phosphorylation and activation of the cdc25C phosphatase[J].Cell Signal, 2000, 12(6):405-411.
24.	Toyoshima-Morimoto F, Taniguchi E, Shinya N, et al.Pololike kinase 1 phosphorylates cyclin B1 and targets it to the nucleus during prophase[J].Nature, 2001, 410(6825):215-220.
25.	范钰,郑树,赵刚.蟾蜍灵对结肠癌SW-480细胞polo-like kinase-1表达及细胞凋亡的影响[J].中国病理生理杂志, 2006, 22(3):491-494.
26.	符芳芳,陈功星,张家炜,等.针对PLK1基因mRNA的SiRNA治疗结肠癌的细胞学研究[J].中国现代应用药学, 2005, 22(z2):699-701.

1. 陈金明,李伟华,王家顿,等.结肠癌细胞凋亡与增殖的平衡及其与预后的关系[J].癌症, 2005, 24(5):554-558.
2. Hörlin A, Krause J, Radü U, et al.Tumor proliferation, MDR phenotype and tumor associated t-lymphocytes in non-Hodgkin's lymphomas[J].Verh Dtsch Ges Pathol, 1992, 76:248-252.
3. Li GC, Zhang Z, Ma XJ, et al.Are biomarkers correlated with recurrence patterns in patients with resectable gastric adenocarcinoma[J].Mol Biol Rep, 2012, 39(1):399-405.
4. Feng W, Malpica A, Skaland I, et al.Can proliferation biomarkers reliably predict recurrence in World Health Organization 2003 defined endometrial stromal sarcoma, low grade?[J].PLoS One, 2013, 8(10):e75899.
5. Hellman K, Johansson H, Andersson S, et al.Prognostic significance of cell cycle-and invasion-related molecular markers and genomic instability in primary carcinoma of the vagina[J].Int J Gynecol Cancer, 2013, 23(1):41-51.
6. Toyoshima-Morimoto F, Taniguchi E, Nishida E.Plk1 promotes nuclear translocation of human Cdc25C during prophase[J].EMBO Rep, 2002, 3(4):341-348.
7. 兰斌,刘炳亚,朱正纲.PLK1在细胞周期生物学的研究进展[J].医学分子生物学杂志, 2005, 2(1):65-67.
8. Katayama H, Sasai K, Kawai H, et al.Phosphorylation by aurora kinase A induces Mdm2-mediated destabilization and inhibition of p53[J].Nat Genet, 2004, 36(1):55-62.
9. Tsai MY, Wiese C, Cao K, et al.A ran signalling pathway mediated by the mitotic kinase Aurora a in spindle assembly[J].Nat Cell Biol, 2003, 5(3):242-248.
10. Sen S, Zhou H, Zhang RD, et al.Amplification/overexpression of a mitotic kinase gene in human bladder cancer[J].J Natl Cancer Inst, 2002, 94(17):1320-1329.
11. Zhou H, Kuang J, Zhong L, et al.Tumour amplified kinase STK15/BTAK induces centrosome amplification, aneuploidy and transformation[J].Nat Genet, 1998, 20(2):189-193.
12. Ouchi M, Fujiuchi N, Sasai K, et al.BRCA1 phosphorylation by Aurora-A in the regulation of G2 to M transition[J].J Biol Chem, 2004, 279(19):19643-19648.
13. 兰斌,陈雪华,刘炳亚,等.STK15基因沉默导致胃癌细胞分裂停滞[J].中华病理学杂志, 2006, 35(2):106-109.
14. Hannak E, Kirkham M, Hyman AA, et al.Aurora-A kinase is required for centrosome maturation in caenorhabditis elegans[J].J Cell Biol, 2001, 155(7):1109-1116.
15. Takahashi T, Sano B, Nagata T, et al.Polo-like kinase 1(PLK1) is overexpressed in primary colorectal cancers[J].Cancer Sci, 2003, 94(2):148-152.
16. Spänkuch-Schmitt B, Bereiter-Hahn J, Kaufmann M, et al.Effect of RNA silencing of polo-like kinase-1(PLK1) on apoptosis and spindle formation in human cancer cells[J].J Natl Cancer Inst, 2002, 94(24):1863-1877.
17. Weichert W, Kristiansen G, Schmidt M, et al.Polo-like kinase 1 expression is a prognostic factor in human colon cancer[J].World J Gastroenterol, 2005, 11(36):5644-5650.
18. 欧海玲,覃宇周,王进声.丝氨酸/苏氨酸激酶15在早期乳腺癌组织中的表达及与人类表皮生长因子受体2、雌激素受体的关系[J].实用医学杂志, 2010, 26(19):3498-3500.
19. 宋波,郝钢跃,马蔚然,等.STK15基因多态性与膀胱癌发病风险的关系[J].现代泌尿外科杂志, 2010, 15(4):277-279.
20. Golsteyn RM, Mundt KE, Fry AM, et al.Cell cycle regulation of the activity and subcellular localization of Plk1, a human protein kinase implicated in mitotic spindle function[J].J Cell Biol, 1995, 129(6):1617-1628.
21. Lee KS, Yuan YL, Kuriyama R, et al.Plk is an M-phase-specific protein kinase and interacts with a kinesin-like protein, CHO1/MKLP-1[J].Mol Cell Biol, 1995, 15(12):7143-7151.
22. Feng Y, Longo DL, Ferris DK.Polo-like kinase interacts with proteasomes and regulates their activity[J].Cell Growth Differ, 2001, 12(1):29-37.
23. Roshak AK, Capper EA, Imburgia C, et al.The human pololike kinase, PLK, regulates cdc2/cyclin B through phosphorylation and activation of the cdc25C phosphatase[J].Cell Signal, 2000, 12(6):405-411.
24. Toyoshima-Morimoto F, Taniguchi E, Shinya N, et al.Pololike kinase 1 phosphorylates cyclin B1 and targets it to the nucleus during prophase[J].Nature, 2001, 410(6825):215-220.
25. 范钰,郑树,赵刚.蟾蜍灵对结肠癌SW-480细胞polo-like kinase-1表达及细胞凋亡的影响[J].中国病理生理杂志, 2006, 22(3):491-494.
26. 符芳芳,陈功星,张家炜,等.针对PLK1基因mRNA的SiRNA治疗结肠癌的细胞学研究[J].中国现代应用药学, 2005, 22(z2):699-701.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Expression and Significance of PLK1 and STK15 Gene, and Effect of Its Specific Inhibitor on Proliferation in Colon Cancer Cells

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