摘要
目的探讨6尿素神经酰胺对结肠癌细胞增殖的影响及其分子生物学机制。方法体外培养人源低分化结肠癌细胞系RKO细胞(购自美国典型菌种保藏中心),经过5(低剂量组)、10(中剂量组)、20μmol/L(高剂量组)不同浓度的6尿素神经酰胺分别诱导处理RKO细胞24、48、72 h,同时设立空白对照组。分别利用噻唑蓝(MTT)、流式细胞术检测各组细胞相对生存率、凋亡率;蛋白免疫印迹(Western blot)检测各组细胞中Wnt信号通路相关蛋白表达情况。采用SPSS 23.0软件中方差分析和t检验进行数据统计学分析。结果随着药物浓度升高和药物作用时间延长,RKO细胞相对生存率[空白组为100%、100%、100%;低剂量组为(85.35±12.52)%、(72.95±10.11)%、(53.55±8.56)%;中剂量组为(70.75±10.74)%、(56.33±7.05)%、(40.05±10.11)%;高剂量组为(50.55±7.42)%、(35.58±5.75)%、(20.98±7.11)%]显著降低(F=23.803,P<0.05),凋亡率显著升高(F=18.284,P<0.05),差异均有统计学意义;各组细胞在相应条件下培养24 h后总蛋白激酶B(Akt)、糖原合成酶激酶3(GSK3β)蛋白表达量差异无统计学意义(F=1.093,P>0.05),但随着药物浓度增加磷酸化Akt、磷酸化GSK3β和β-连环蛋白(β-catenin)表达量显著降低(F=6.824,P<0.05),差异有统计学意义。结论6尿素神经酰胺可以通过抑制Akt磷酸化,从而抑制GSK3β磷酸化,减少β-catenin表达,抑制结肠癌细胞增殖。
Objective To explore the effect of 6-urea-ceramide on the proliferation of colon cancer cells and its molecular biological mechanism.Methods Different concentrations of 6-urea-ceramide were used to treat the low differentiated colon cancer cell line RKO cells for 24,48 and 72 h,and then the relative survival rate and apoptosis rate were detected.Finally,the expression of Wnt signaling pathway related proteins in each group was detected.Results With the increase of drug concentration and the prolongation of drug action time,the relative survival rate of RKO cells[blank group:100%,100%,100%;low-dose group:(85.35±12.52)%,(72.95±10.11)%,(53.55±8.56)%;medium-dose group:(70.75±10.74)%,(56.33±7.05)%,(40.05±10.11)%;high-dose group:(50.55±7.42)%,(35.58±5.75)%,(20.98±7.11)%]was significantly reduced(F=23.803,P<0.05),apoptosis rate was significantly increased(F=18.284,P<0.05).After the RKO cells were cultured for 24 h under corresponding conditions,the total protein kinase B(Akt)and glycogen synthase kinase-3β(GSK3β)showed no significant difference(F=1.093,P>0.05).The expression of phosphorylated Akt,phosphorylated GSK3βandβ-catenin decreased significantly with the increase of drug concentration(F=6.824,P<0.05).Conclusion 6-urea-ceramide can inhibit the phosphorylation of GSK3β,decrease the expression ofβ-catenin and inhibit the proliferation of colon cancer cells by inhibiting Akt phosphorylation.
作者
毕于合
曹帅
李涛
魏胜超
任维才
刘洪俊
Bi Yuhe;Cao Shuai;Li Tao;Wei Shengchao;Ren Weicai;Liu Hongjun(Department of Anorectal Surgery,Jinan People’s Hospital,Jinan 271199,China;Department of General Surgery,Shandong Provincial Hospital,Jinan 250021,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2020年第6期1086-1088,共3页
Chinese Journal of Experimental Surgery
