Feng Xie Shun, Ming Sun Shu, Feng Lu Xiao, Feng Xie Ze
Departments of General Surgery, The First Affiliated Hospital of Shantou University Medical College, Shantou, People's Republic of China.
Mol Biol Rep. 2014 Feb;41(2):987-96. doi: 10.1007/s11033-013-2943-2. Epub 2014 Jan 9.
In the study presented here, we first evaluated effect of CDDP on liver cancer cells SMMC-7721 apoptosis and motility capacity. Then, we evaluate inhibitory effect of CDDP on tumour growth and its possible molecular mechanism in liver cancer mice model. Results showed that the apoptosis rate of cells decreased with increasing CDDP. Analysis of the effect of the CDDP on cell cycle was performed by flow cytometry and results show a dose-dependent increase in the percentage of cells in the S-phase of the cell cycle, with a decrease in the percentage of cells in the G1 and G2/M phases. CDDP did not close the wound even after 48 h, as opposed to untreated cells (0 mg/l). Similarly, the migratory and invasion capacity of SMMC-7721 cells was also reduced after treatment with CDDP, as evaluated by a transwell assay. Animal experiment indicated that CDDP administration could increase blood WBC, total protein, albumin and A/G, decrease blood alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase levels in hepatocellular carcinomas mice. Immunohistochemistry analysis showed that positive expression of Fas and Bax proteins in the medicine-treated (II, III) group was significantly higher, whereas the expression of NF-κB, P53, Bcl-2 proteins was significantly lower than those of the control group. Gene expression analysis using Real time PCR methods revealed a significant up-regulation in the expression levels of Bax mRNA in the medicne-treated (II, III) group when compared to untreated control. In contrast, CDDP-treated group showed a significant down regulation in the expression levels of Bcl-2 mRNA as compared to untreated control group. These results are in agreement with immunohistochemistry data. Our observations indicate that CDDP has damaged effects on liver tumour cells SMMC-7721 including apoptosis, motility and cell cycle under in vitro. CDDP can enhance pro-apoptosis gene Fas, Bax expression, decrease anti-apoptosis genes Bcl-2 expression, and mutant genes P53, NF-κB proteins expression.
在本研究中，我们首先评估了顺铂对肝癌细胞SMMC - 7721凋亡和运动能力的影响。然后，我们评估了顺铂对肝癌小鼠模型中肿瘤生长的抑制作用及其可能的分子机制。结果显示，随着顺铂浓度增加，细胞凋亡率降低。通过流式细胞术分析顺铂对细胞周期的影响，结果显示细胞周期S期细胞百分比呈剂量依赖性增加，而G1期和G2/M期细胞百分比降低。与未处理的细胞（0 mg/l）相比，即使在48小时后顺铂处理组也未封闭伤口。同样，通过Transwell实验评估，顺铂处理后SMMC - 7721细胞的迁移和侵袭能力也降低。动物实验表明，给予顺铂可使肝癌小鼠血液中的白细胞、总蛋白、白蛋白和白球比升高，丙氨酸氨基转移酶、天冬氨酸氨基转移酶和碱性磷酸酶水平降低。免疫组化分析显示，药物处理组（II、III）中Fas和Bax蛋白的阳性表达显著高于对照组，而NF - κB、P53、Bcl - 2蛋白的表达显著低于对照组。使用实时PCR方法进行基因表达分析显示，与未处理的对照组相比，药物处理组（II、III）中Bax mRNA的表达水平显著上调。相反，与未处理的对照组相比，顺铂处理组中Bcl - 2 mRNA的表达水平显著下调。这些结果与免疫组化数据一致。我们的观察表明，顺铂在体外对肝癌细胞SMMC - 7721具有损伤作用，包括凋亡、运动和细胞周期方面。顺铂可增强促凋亡基因Fas、Bax的表达，降低抗凋亡基因Bcl - 2的表达，以及突变基因P53、NF - κB蛋白的表达。
