摘要
目的探索氨苄西林通过调控泛素样含PHD和环指域蛋白2(UHRF2)和组蛋白H3赖氨酸残基(H3K9)甲基化水平的抗结肠癌作用机制。方法将SW480、HT29细胞均分为对照组和实验组。实验组给予含1μg·mL^(-1)氨苄西林的RPMI培养基进行培养;对照组给予不含氨苄西林的RPMI培养基进行培养。用蛋白质印迹法检测UHRF2和H3K9甲基化水平,用CCK-8实验法检测细胞的增殖能力,用划痕实验法检测细胞的迁移能力,用Transwell实验法检测细胞的侵袭能力。结果在SW480细胞中,实验组和对照组的UHRF2蛋白相对表达水平分别为102.36±10.12和698.67±36.92,H3K9二甲基化(H3K9me2)蛋白相对表达水平分别为406.59±20.97和721.52±36.20,H3K9三甲基化(H3K9me3)蛋白相对表达水平分别为219.78±25.88和821.97±50.61,细胞凋亡率分别为(45.28±6.35)%和(26.18±3.25)%,细胞划痕愈合率分别为(9.47±3.54)%和(26.29±4.29)%,侵袭细胞数分别为(29.23±3.78)和(39.52±4.15)个,差异均有统计学意义(均P<0.05)。在HT29细胞中,实验组和对照组的UHRF2蛋白相对表达水平分别为160.85±29.74和551.79±39.76,H3K9me2蛋白相对表达水平分别为385.26±46.98和601.30±29.86,H3K9me3蛋白相对表达水平分别为496.85±36.22和725.91±53.62,细胞凋亡率分别为(50.35±7.43)%和(29.45±5.65)%,细胞划痕愈合率分别为(6.54±4.16)%和(21.14±5.74)%,侵袭细胞数分别为(32.22±3.62)和(41.63±5.59)个,差异均有统计学意义(均P<0.05)。结论氨苄西林可能通过下调UHRF2蛋白水平及H3K9甲基化水平,从而抑制结肠癌细胞的生长。
Objective To investigate the mechanism of ampicillin against colon cancer by regulating the expression of ubiquitin-like with PHD and ring finger domains 2(UHRF2)and the methylation level of related histone H3 K9(histone H3 lysine residue).Methods SW480 and HT29 cells were divided into control group and experimental group,respectively.The experimental group was cultured with 1μg·mL^(-1)ampicillin in RPMI medium and the control group was cultured in RPMI medium without ampicillin.The levels of UHRF2 and H3 K9 methylation were measured by Western blot.CCK-8 assay,scratch assay and Transwell assay were used to detect the effects on cell proliferation,migration and invasion,respectively.Results In SW480 cells,the relative expression levels of UHRF2 protein in the experimental and control groups were 102.36±10.12 and 698.67±36.92,the relative expression levels of di-methylation at H3K9(H3K9me2)protein were 406.59±20.97 and 721.52±36.20,the relative expression levels of tri-methylation at H3K9(H3K9me3)protein were 219.78±25.88 and 821.97±50.61,the apoptosis rates were(45.28±6.35)%and(26.18±3.25)%,the cell scratch healing rates were(9.47±3.54)%and(26.29±4.29)%,the number of invaded cells was 29.23±3.78 and 39.52±4.15,and the differences were statistically significant(all P<0.05).In HT29 cells,the relative expression levels of UHRF2 protein in the experimental and control groups were 160.85±29.74 and 551.79±39.76,the relative expression levels of H3K9me2 protein were 385.26±46.98 and 601.30±29.86,the relative expression levels of H3K9me3 protein were496.85±36.22 and 725.91±53.62,the apoptosis rates were(50.35±7.43)%and(29.45±5.65)%,the cell scratch healing rates were(6.54±4.16)%and(21.14±5.74)%,the number of invasive cells was 32.22±3.62and 41.63±5.59,and the differences were statistically significant(all P<0.05).Conclusion Ampicillin may inhibit the growth of colon cancer cells by down-regulating UHRF2 protein levels and H3K9 methylation levels.
作者
陈思圜
孙菁婕
方娴韵
吴可佳
段昌柱
CHEN Si-yuan;SUN Jing-jie;FANG Xian-yun;WU Ke-jia;DUAN Chang-zhu(School of Basic Medicine,Chongqing Medical University,Chongqing 400016,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2022年第23期2845-2849,共5页
The Chinese Journal of Clinical Pharmacology
基金
国家自然科学基金资助项目(8177080705)。
