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目的探讨沙利度胺单药及联合应用5-氟尿嘧啶(5-FU)+奥沙利铂对肝癌细胞株SMMC7721生长的影响。方法采用MTT法检测确定各个单药的IC50,并以药物的IC50作为后续实验的给药浓度。实验分为空白对照组、沙利度胺组、5-FU+奥沙利铂组及沙利度胺联合5-FU+奥沙利铂组。MTT法检测各组药物作用24 h及48 h后对肝癌细胞的抑制率。各组药物作用48 h后,采用流式细胞术检测细胞周期,RT-PCR法测定SMMC7721细胞的Bcl-2、Bax mRNA的表达,Western blot法分析Bcl-2、Bax蛋白的表达。结果 MTT结果显示,药物作用24、48 h后,沙利度胺能够明显抑制肝癌细胞SMMC7721的增殖,促进其凋亡(抑制率分别为14.86%和24.24%),而沙利度胺联合5-FU+奥沙利铂的抑制作用更加显著(抑制率分别为29.80%和40.53%),呈现时间依赖性。药物作用48h后,三个用药组的肝癌细胞均被阻滞于G0/GI期,且沙利度胺联合5-FU+奥沙利铂组的阻滞作用更为明显(P<0.05)。沙利度胺下调Bcl-2基因表达而上调Bax基因表达,且沙利度胺联合5-FU+奥沙利铂对Bcl-2、Bax基因的调控作用更为显著,差异均有统计学意义(P<0.01)。结论沙利度胺能够明显抑制SMMC7721细胞的增殖,且与5-FU及奥沙利铂具有协同作用,其机制可能与影响Bcl-2及Bax的表达有关。
Objective To investigate the effects of thalidomide monotherapy combined with 5-fluorouracil (5-FU) and oxaliplatin on the growth of hepatocellular carcinoma cell line SMMC7721. Methods The IC50 of each single drug was determined by MTT assay, and the IC50 of the drug was used as the drug concentration for subsequent experiments. The experiment was divided into blank control group, thalidomide group, 5-FU + oxaliplatin group and thalidomide combined with 5-FU + oxaliplatin group. The inhibition rate of hepatoma cells after 24 h and 48 h of each group was detected by MTT assay. The cell cycle was detected by flow cytometry after 48 h of each drug treatment. The expression of Bcl-2 and Bax mRNA was detected by RT-PCR and the expression of Bcl-2 and Bax by Western blot. Results MTT showed that thalidomide could significantly inhibit the proliferation and promote the apoptosis of hepatocellular carcinoma SMMC7721 cells 24 h and 48 h after treatment with the inhibitory rates of 14.86% and 24.24%, respectively. Thalidomide combined with 5- The inhibitory effect of FU + oxaliplatin was more significant (the inhibitory rates were 29.80% and 40.53%, respectively), showing a time-dependent manner. After 48 hours of drug treatment, the hepatoma cells in all three groups were blocked in G0 / GI phase, and the blocking effect of thalidomide combined with 5-FU + oxaliplatin group was more obvious (P <0.05). Thalidomide down-regulated Bcl-2 gene expression and up-regulated Bax gene expression, and the effect of thalidomide combined with 5-FU + oxaliplatin on Bcl-2 and Bax gene expression was more significant, the difference was statistically significant P <0.01). Conclusions Thalidomide can significantly inhibit the proliferation of SMMC7721 cells and has a synergistic effect with 5-FU and oxaliplatin. The mechanism may be related to the influence of Bcl-2 and Bax expression.