维生素D类似物EB1089对肝癌细胞生长抑制作用的研究

The study of vitamin D analogue EB1089 on the proliferation of hepatocellular carcinoma cells

目的探讨维生素D类似物EB1089对肝癌细胞在体外和体内生长环境下的抑制作用及其可能机制。方法①体外培养肝癌SMMC-7721细胞株,以1、10、100、1 000nmol/L的EB1089分别作用24、48和72 h,采用MTT法测定EB1089对细胞生长增殖的抑制作用;用RT-PCR法检测长链脂肪酸辅酶A连接酶3(FACL3)、长链脂肪酸辅酶A连接酶5(FACL5)的mRNA表达。②将肝癌SMMC-7721细胞悬液皮下注射于BALB/c Nu/Nu裸鼠,建立人肝癌裸鼠皮下移植瘤模型,随机分组后实验组分别给予0.5和1.0μg/(kg.d)腹腔注射,对照组给予等量乙醇溶液,不处死裸鼠情况下每5 d计量肿瘤大小,绘制生长曲线图。结果在体外与体内环境下,EB1089均可以抑制肝癌细胞株的生长增殖。同时,EB1089下调SMMC-7721细胞中FACL5 mRNA的表达,但是对FACL3 mRNA的表达则无明显影响。结论 EB1089可能通过FACL5途径抑制肝癌细胞的生长增殖。

Objective To investigate the inhibitory effect of vitamin D analogue EB1089 on the proliferation of hepatocellular carcinoma cells in vitro and vivo culture and its possible mechanism.Methods ① SMMC-7721 hepatocellular carcinoma cell lines were exposed to EB1089 at 1,10,100,1 000 nmol / L for 24,48 and 72 h,the inhibitory rate of cells was detected by MTT assay,the expression of long-chain fatty-acid-CoA ligase3（FACL3）,longchain fatty-acid-CoA ligase 5（FACL 5） mRNA in cells were detected by using RT-PCR methods;② for in vivo analysis,SMMC-7721 hepatocellar carcinoma cells were xenografted into BALB / c Nu / Nu nude mice,mice were randomly assigned to a control group（n = 10） or one of the threatment groups（2 groups of 10 mice） receiving 0.5 or 1 μg /（kg.d） of EB1089.Control animals received equal volume vehicle（ethanol）.Tumor size was measured every five day without killing and the tumor growth curve was made.Results Proliferation of hepatocellar cells were significantly inhibited at EB1089 concentrations tested both in vitro culture.Furthermore,EB1089 increased the expression of FACL5 mRNA while there were not significant change in the expression of FACL3 mRNA.Conclusion EB1089 may inhibit the proliferation of hepatocellular carcinoma cell lines via FACL5.