Ren Kai, Ma Linyan, Chong Dianlong, Zhang Zhirui, Zhou Can, Liu Hao, Zhao Surong
Department of Pharmacy, Bengbu Medical College; Anhui Engineering Technology Research Center of Biochemical Pharmaceutics, Bengbu Anhui 233030, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2016 Sep 28;41(9):898-904. doi: 10.11817/j.issn.1672-7347.2016.09.003.
To investigate the effects of LCL161, a Smac mimetic, on the proliferation and apoptosis in hepatocellular carcinoma cells and the underlying mechanisms.
The effect of LCL161 on the cell viability of HepG2 and SMMC7721 cells was measured by MTT assay. The effect of LCL161 at lower concentrations on the proliferation in hepatocellular carcinoma (HCC) cells was detected by colony formation assay. Apoptosis was assessed by flow cytometry with PI staining. The mitochondrial membrane potential was measured by JC-1 staining. The expression of PARP, p-Akt, cIAP1 and XIAP protein was analyzed by Western blot.
LCL161 displayed notable antiproliferative activity on HCC cells at the concentrations of 1-16 μmol/L (P<0.01), with IC50 values of 4.3 and 4.9 μmol/L for HepG2 and SMMC7721 cells, respectively, after treatment for 48 h. LCL161 at lower concentrations obviously inhibited the colony formation of HCC cells. LCL161 induced significant apoptosis in HCC cells (P<0.01), and resulted in the apoptotic rate at (1.5±0.8)% or (1.8±0.6)% , (15.2±2.8)% or (12.2±2.4)%, (28.7±3.0)% or (22.4±2.7)%, (34.6±2.3)% or (30.2±2.4)% for HepG2 cells or SMMC7721 cells at the concentration of 0, 2, 4 or 8 μmol/L, respectively. The result of JC-1 staining indicated that the mitochondrial membrane potential of HCC cells was reduced by LCL161. In addition, LCL161 promoted the cleavage of PARP, down-regulated the protein expression of p-Akt, and degraded cIAP1.
LCL161 possesses significant anti-proliferative activity and pro-apoptotic action in HepG2 and SMMC7721 cells, which might be correlated with reduction in mitochondrial membrane potential, down-regulation of p-Akt and degradation of cIAP1.
研究Smac模拟物LCL161对肝癌细胞增殖和凋亡的影响及其潜在机制。
采用MTT法检测LCL161对HepG2和SMMC7721细胞活力的影响。采用集落形成试验检测低浓度LCL161对肝癌（HCC）细胞增殖的影响。通过PI染色流式细胞术评估细胞凋亡。采用JC-1染色检测线粒体膜电位。通过蛋白质印迹法分析PARP、p-Akt、cIAP1和XIAP蛋白的表达。
LCL161在1-16μmol/L浓度下对HCC细胞显示出显著的抗增殖活性（P<0.01），处理48小时后，HepG2和SMMC7721细胞的IC50值分别为4.3和4.9μmol/L。低浓度的LCL161明显抑制HCC细胞的集落形成。LCL161诱导HCC细胞显著凋亡（P<0.01），在0、2、4或8μmol/L浓度下，HepG2细胞或SMMC7721细胞的凋亡率分别为（1.5±0.8）%或（1.8±0.6）%、（15.2±2.8）%或（12.2±2.4）%、（28.7±3.0）%或（22.4±2.7）%、（34.6±2.3）%或（30.2±2.4）%。JC-1染色结果表明LCL161降低了HCC细胞的线粒体膜电位。此外，LCL161促进PARP的裂解，下调p-Akt的蛋白表达，并降解cIAP1。
LCL161在HepG2和SMMC7721细胞中具有显著的抗增殖活性和促凋亡作用，这可能与线粒体膜电位降低、p-Akt下调和cIAP1降解有关。
