PLK1及STK15基因在结肠癌细胞中的表达及其特异性抑制剂对结肠癌细胞增殖的影响

Expression and Signifi cance of PLK1 and STK15 Gene, and Effect of Its Specifi c Inhibitor on Proliferation in Colon Cancer Cells

目的研究保罗样激酶1(polo-like kinase 1,PLK1)及丝氨酸/苏氨酸激酶15(serine/threonine kinase15,STK15)mRNA及其蛋白在结肠癌细胞中的表达,以及其特异性抑制剂对结肠癌细胞增殖的影响。方法选取人宫颈癌Hela细胞和人结肠癌HCT-116细胞、HT-29细胞及CACO-2细胞,以β-actin为内参照,采用逆转录聚合酶链反应(RT-PCR)法检测PLK1 mRNA和STK15 mRNA的表达水平;采用Western blot法检测PLK1蛋白和STK15蛋白的表达水平;采用偶氮唑盐(MTT)法检测Dulbecco改良细胞培养基(DMEM培养基)、二甲基亚砜(DMSO)、SBE13(PLK1蛋白特异性抑制剂)或VX-680(STK15蛋白特异性抑制剂)处理后4种癌细胞的增殖活性。结果与Hela细胞比较,HCT-116细胞、HT-29细胞及CACO-2细胞的PLK1和STK15 mRNA及其蛋白的表达水平均较高(P<0.05)。①Hela细胞:与DMEM组比较,SBE13组、VX-680组及SBE13+VX-680组的增殖能力无明显变化(P>0.05)。②HCT-116细胞和HT-29细胞:与DMEM组比较,VX-680组和SBE13+VX-680组的增殖能力均降低(P<0.05),但SBE13组的差异无统计学意义(P>0.05)。③CACO-2细胞:与DMEM组比较,SBE13组、VX-680组及SBE13+VX-680组的增殖能力均降低(P<0.05)。结论 HCT-116、HT-29及CACO-2结肠癌细胞的PLK1 mRNA和STK15 mRNA及其蛋白的表达水平均高于Hela细胞,应用其特异性抑制剂可抑制部分结肠癌细胞系的生长。

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, STK15mRNA 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 methylthiazolyldi-phenyl-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）. ①Hela 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）. ②HCT-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）. ③CACO-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.