SETDB1通过p53/p21通路调控结肠癌细胞衰老  

SETDB1 regulates cell senescence through p53/p21 signaling

作　　者：吴倩[1] 侯吉学[1] 贺强 崔晓宾[2] 黄桂林[1] 孙旭凌[1] Wu Qian;Hou Jixue;He Qiang;Cui Xiaobin;Huang Guilin;Sun Xuling(Dept of Gastrointestinal and Breast Surgery,The First Affiliated Hospital of Medical College of Shihezi University,Shihezi 832008;Dept of Pathology,School of Medicine,Shihezi University,Shihezi 832008)

机构地区：[1]石河子大学医学院第一附属医院胃肠乳腺外科,石河子832008 [2]石河子大学医学院病理教研室,石河子832008

出　　处：《安徽医科大学学报》2022年第9期1442-1446,1452,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金-新疆联合基金(编号:U1903305);石河子大学创新发展专项基金(编号:CXFZ202113);石河子大学医学院第一附属医院博士基金(编号:BS202102)。

摘　　要：目的探讨组蛋白甲基化酶SET结构域分支型1(SETDB1)对结肠癌细胞衰老与增殖影响。方法利用多柔比星(DOX)诱导结肠癌细胞衰老,Western blot证实SETDB1在衰老细胞中的蛋白表达量。利用SETDB1的shRNA(shSETDB1#1和shSETDB1#2)和过表达质粒分别构建低表达和过表达SETDB1的结肠癌细胞模型,用Western blot检测SETDB1表达程度;用CCK8法和β-半乳糖苷酶染色法分别检测细胞增殖能力及衰老细胞的百分比,p53和p21的变化由Western blot检测;针对p53过表达质粒行逆转实验。结果DOX诱导细胞衰老后SETDB1的蛋白表达量可降低。低表达SETDB1后结肠癌细胞的增殖能力下降,衰老细胞比例由(22.00±4.35)%升至(54.00±5.56)%和(53.33±4.93)%(P<0.001);过表达SETDB1后结肠癌细胞增殖能力升高,衰老细胞比例从(43.33±6.11)%降低至(21.33±3.51)%(P<0.01);通过GSE56496数据富集分析发现SETDB1与p53信号通路有关,沉默SETDB1可增高p53和p21蛋白水平,而过表达SETDB1可降低p53和p21蛋白水平;过表达p53可逆转过表达SETDB1引起的细胞衰老减少现象。结论SETDB1通过下调p53/p21信号通路抑制结肠癌细胞衰老。Objective To investigate the effects of SETDB1on the proliferation and apoptosis of human colon cancer cells.Methods The senescence of colon cancer cells was induced by doxorubicin,and the protein expression of SETDB1 in the senescent cells was detected by Western blot.Colon cancer cell models with low expression of SETDB1 and overexpression of SETDB1 were constructed by using shRNA(shSETDB1#1 and shSETDB1#2)of SETDB1 and overexpression plasmid,respectively.Western blot was used to detect the expression of SETDB1,the CCK8 assay was performed to detect cell proliferation while a Senescenceβ-Galactosidase Staining Kit was used to detect senescent cells,and Western blot was used to detect the changes of p53 and p21.The reverse experiment was carried out with p53 overexpression plasmid.Results Cell senescence evoked by doxorubicin could down-regulate the protein expression of SETDB1.After low expression of SETDB1,the proliferation ability of colonic cancer cells decreased,and the proportion of senescent cells increased from(22.00±4.35)%to(54.00±5.56)%and(53.33±4.93)%(P<0.001).After overexpression of STEDB1,the proliferation ability of colon cancer cells increased,and the ratio of senescent cells decreased from(43.33±6.11)%to(21.33±3.51)%(P<0.01).Through GSE56496 enrichment and analysis,it was found that SETDB1 was related to p53 signal pathway,silencing SETDB1 could increase the levels of p53 and p21 protein,while overexpression of SETDB1 could decrease the level of p53 and p21 protein,and overexpression of p53 reversed the decrease of cell senescence caused by overexpression of SETDB1.Conclusion SETDB1 inhibits the senescence of colon cancer cells by inhibiting p53/p21 signal pathway,which provides a potential target for individualized tumor therapy.

关 键 词：结肠癌 SETDB1 细胞增殖 细胞衰老 P53 

分 类 号：R735.3[医药卫生—肿瘤]