摘要目的:探讨靶向抑制Polo样激酶1（PLK1）对子宫颈癌细胞增殖、有丝分裂、细胞凋亡的影响及可能机制。方法:取对数生长期人子宫颈癌HeLa、C-33A细胞株，以10、20 nmol/L PLK1抑制剂GSK461364处理的细胞作为不同浓度GSK461364组，以未经GSK461364处理的细胞作为对照组。采用CCK-8法检测细胞增殖能力（以450 nm波长处吸光度值表示），流式细胞术检测细胞染色体倍性（碘化丙啶染色），通过流式细胞术检测的线粒体膜电位评估细胞凋亡状态（JC-1荧光探针，绿色荧光的JC-1单体所在细胞为凋亡细胞），蛋白质印迹法检测细胞周期、细胞凋亡相关蛋白的表达水平。结果:CCK-8法检测结果显示，10、20 nmol/L GSK461364处理24 h后去除GSK461364继续培养24、48、72 h后HeLa细胞增殖能力均低于对照组，10、20 nmol/L GSK461364处理24 h后去除GSK461364继续培养48、72 h后C-33A细胞增殖能力均低于对照组，差异均有统计学意义（均 P<0.05）。流式细胞术检测结果显示，GSK461364作用24 h去除后继续培养72 h，10、20 nmol/L GSK461364组和对照组HeLa细胞中多倍体细胞亚群比例分别为（13.89±3.73）%、（12.30±5.49）%、（9.86±1.15）%，差异无统计学意义（ F=0.82， P>0.05）；10、20 nmol/L GSK461364组和对照组C-33A细胞中多倍体细胞亚群比例分别为（8.45±2.20）%、（11.06±2.53）%、（5.42±1.36）%，差异有统计学意义（ F=5.46， P=0.045），其中，20 nmol/L GSK461364组多倍体细胞亚群比例高于对照组，差异有统计学意义（ t=3.40， P=0.027）。流式细胞术检测线粒体膜电位结果显示，GSK461364作用24 h去除后继续培养72 h，对照组、10 nmol/L GSK461364组、20 nmol/L GSK461364组HeLa细胞中凋亡细胞比例分别为（3.96±2.28）%、（24.38±4.89）%、（46.24±4.38）%，差异有统计学意义（ F=83.18， P<0.000 1），10、20 nmol/L GSK461364组凋亡细胞比例均高于对照组，差异均有统计学意义（均 P<0.01），且20 nmol/L组凋亡细胞比例高于10 nmol/L组（ t=5.76， P=0.005）；对照组、10 nmol/L GSK461364组、20 nmol/L GSK461364组C-33A细胞中凋亡细胞比例分别为（1.81±1.59）%、（5.22±1.57）%、（15.87±5.81）%，差异有统计学意义（ F=12.49， P=0.007），且20 nmol/L组凋亡细胞比例高于10 nmol/L组和对照组（均 P<0.05）。蛋白质印迹法检测结果显示，GSK461364作用24 h去除后继续培养72 h，10、20 nmol/L GSK461364组HeLa和C-33A细胞裂解型胱天蛋白酶9、裂解型多腺苷二磷酸核糖聚合酶相对表达水平均高于对照组，cdc25c、磷酸化cdc25c（Ser216）相对表达水平均低于对照组，差异均有统计学意义（均 P<0.05）。 结论:靶向抑制PLK1能够体外抑制子宫颈癌细胞的增殖活性，诱导细胞有丝分裂周期阻滞，促进细胞凋亡；这可能是通过调节细胞周期和细胞凋亡相关蛋白实现的。

abstractsObjective:To investigate the effects and possible mechanisms of targeted inhibition of Polo-like kinase 1 (PLK1) on the proliferation, mitosis and apoptosis of cervical cancer cells.Methods:Logarithmically growing human cervical cancer cell lines HeLa and C-33A were selected, and cells treated with 10 and 20 nmol/L PLK1 inhibitor GSK461364 were used as different concentrations of GSK461364 groups, while cells not treated with GSK461364 were used as the control group. CCK-8 method was used to detect cell proliferation ability (represented by absorbance values at wavelength 450 nm), flow cytometry was used to detect chromosome ploidy (propidium iodide staining), mitochondrial membrane potential detected by flow cytometry was used to evaluate cell apoptosis status (JC-1 fluorescent probe, the cells where the JC-1 monomers emitting green fluorescence were located were apoptotic cells), and Western blotting was used to detect the expression levels of cell cycle and apoptosis-related proteins.Results:The results of CCK-8 method showed that the proliferation ability of HeLa cells was lower than that of the control group after 24 hours of treatment with 10 and 20 nmol/L GSK461364 and continued culture for 24, 48 and 72 hours without GSK461364. The proliferation ability of C-33A cells was lower than that of the control group after 24 hours of treatment with 10 and 20 nmol/L GSK461364 and continued culture for 48 and 72 hours without GSK461364, and the differences were statistically significant (all P < 0.05). The results of flow cytometry analysis showed that after 24 hours of treatment with GSK461364 and continued culture for 72 hours without GSK461364, the proportions of polyploid cell subpopulations in HeLa cells of the 10 and 20 nmol/L GSK461364 groups and the control group were (13.89±3.73)%, (12.30±5.49)% and (9.86±1.15)%, respectively, with no statistically significant difference ( F = 0.82, P > 0.05); the proportions of polyploid cell subpopulations in C-33A cells of the 10 and 20 nmol/L GSK461364 groups and the control group were (8.45±2.20)%, (11.06±2.53)% and (5.42±1.36)%, respectively, with statistically significant difference ( F = 5.46, P = 0.045). Among them, the proportion of polyploid cell subpopulations in the 20 nmol/L GSK461364 group was higher than that in the control group, with statistically significant differences ( t = 3.40, P = 0.027). The results of flow cytometry detection of mitochondrial membrane potential showed that after 24 hours of treatment with GSK461364 and continued culture for 72 hours without GSK461364, the proportions of apoptotic cells in HeLa cells of the control group, 10 nmol/L GSK461364 group and 20 nmol/L GSK461364 group were (3.96±2.28)%, (24.38±4.89)%, and (46.24±4.38)%, respectively, and the difference was statistically significant ( F = 83.18, P < 0.000 1), the proportion of apoptotic cells in the 10 and 20 nmol/L GSK461364 groups was higher than that in the control group, and the difference was statistically significant (both P < 0.01), and the proportion of apoptotic cells in the 20 nmol/L group was higher than that in the 10 nmol/L group ( t = 5.76, P = 0.005); the proportions of apoptotic cells in C-33A cells of the control group, 10 nmol/L GSK461364 group and 20 nmol/L GSK461364 group were (1.81±1.59)%, (5.22±1.57)% and (15.87±5.81)%, respectively, with statistically significant differences ( F = 12.49, P = 0.007), and the proportion of apoptotic cells in the 20 nmol/L group was higher than that in the 10 nmol/L group and the control group (both P < 0.05). The results of Western blotting analysis showed that after 24 hours of treatment with GSK461364 and continued culture for 72 hours without GSK461364, the relative expression levels of cleaved Caspase-9 and cleaved polyadenosine diphosphate-ribose polymerase in HeLa and C-33A cells treated with 10 and 20 nmol/L GSK461364 were higher than those in the control group, and the relative expression levels of cdc25c and phosphorylated cdc25c (Ser216) were lower than those in the control group, and the differences were statistically significant (all P < 0.05). Conclusions:Targeted inhibition of PLK1 can inhibit the proliferation activity of cervical cancer cells in vitro, induce cell mitotic cycle arrest, and promote cell apoptosis; these may be achieved by regulating cell cycle and apoptosis-related proteins.