CDK4/6抑制剂阿贝西利联合顺铂对人胃癌细胞生物学行为的影响

Effects of CDK4/6 inhibitor abemaciclib and cisplatin on biological behavior of human gastric cancer cells

目的:探讨CDK4和CDK6(CDK4/6)抑制剂阿贝西利与顺铂联合作用对人胃癌细胞生物学行为的影响。方法:利用生物信息学网站UALCAN在线分析TCGA数据库,获取人胃癌组织与正常胃组织中CDK4/6基因表达情况;培养人胃癌细胞SGC-7901,采用Western blot检测细胞中CDK4/6蛋白的表达。利用细胞增殖实验检测顺铂和阿贝西利单独作用24 h的半数抑制浓度,然后将细胞分为空白对照组、0.5μg/mL顺铂组、10μmol/L阿贝西利组、10μmol/L阿贝西利和0.5μg/mL顺铂联合组,分别作用24、48、72 h后采用CCK-8法、流式细胞术、划痕实验、Transwell实验检测各组中SGC-7901细胞的增殖、凋亡、迁移和侵袭情况。结果:生物信息学分析表明CDK4/6蛋白在人胃癌组织中的表达显著高于正常胃组织(P<0.05)。Western blot实验显示,与对照组相比,人胃癌细胞SGC-7901中CDK4/6蛋白的表达水平均显著升高(P<0.05)。CCK-8实验结果显示最接近IC50值的顺铂浓度为0.5μg/mL,阿贝西利浓度为10μmol/L,故后续选择此浓度进行试验。与空白对照组、顺铂组、阿贝西利单独组相比,阿贝西利联合顺铂组的细胞增殖抑制率明显增强且具有时间依赖性(r=0.949,P<0.01),细胞凋亡率亦显著升高(P<0.01),迁移和侵袭能力均显著减弱(P<0.01)。结论:CDK4/6在人胃癌组织和细胞中的表达均显著升高,CDK4/6抑制剂阿贝西利联合顺铂显示出协同作用,可显著抑制人胃癌细胞的增殖、迁移和侵袭,并促进其凋亡。

OBJECTIVE:To investigate biological behavior of human gastric cancer cells after their treatment with the CDK4 and CDK6(CDK4/6)inhibitor abemaciclib,and cisplatin.METHODS:The TCGA database resources and the bioinformatics UALCAN website were used to obtain the expression of CDK4/6 genes in gastric cancers and normal gastric tissues.Western blot test was used to detect expression of CDK4 and CDK6 albumen in the stomach carcinoma cell line SGC7901 and the regular gastric mucous membrane cell line GES-1.The CCK-8 method was used to screen for the optimal cell growth inhibition concentration.The cells were divided into several groups:blank control,0.5μg/mL cisplatin,10μmol/L abemaciclib+0.5μg/mL cisplatin,and 10μmol/L abemaciclib groups.These groups were treated for 24,48 and 72 h.Then CCK-8 method,flow cytometry,scratch assay and Transwell assay were used to detect the proliferation,apoptosis,migration and invasion gastric cancer cells SGC-7901 in each subgroup RESULTS:Based on bioinformatics analysis,expression of CDK4/6 proteins in human gastric cancer tissues was significantly higher than that in normal gastric tissues(P<0.05).Western blot analyses showed that expression of CDK4/6 proteins was significantly increased in SGC7901 compared with that in normal tissues(P<0.05).According to the CCK-8 results,the cisplatin concentration for the IC50 value was 0.5μg/mL and the abemaciclib was 10μmol/L,therefore these concentrations were used for subsequent experiments.Compared with the blank control group,the combined treatment inhibited cell proliferation significantly(P<0.01),and the inhibition ability was time-dependent(r=0.949,P<0.01).The level of apoptosis was significantly increased(P<0.01),and both migration and invasion capacity were significantly reduced(P<0.01).CONCLUSION:Expression of CDK4/6 was significantly increased in human gastric cancer tissues and cells.Combined treatment of these cells with abemaciclib and cisplatin showed synergistic effects,which significantly inhibited the proliferation,migration,invasion of human gastric cancer cells and promoted apoptosis.