摘要目的 探讨柠檬酸镧对肝癌细胞增殖、凋亡的影响及其可能的机制.方法 分别使用不同浓度的环巴胺-KAAD以及柠檬酸镧处理人肝癌细胞株SMMC-7721.采用MTT法检测细胞增殖,采用Annexin V/PI流式细胞术检测细胞凋亡率.采用蛋白印迹(Western blot)检测CyclinD1、p21、Caspase-3、bcl-2、Gli1、Shh表达.采用实时定量(q-PCR)检测Gli1、Shh的表达.结果 对照组、0.1 mmol/L柠檬酸镧组、15 μmol/L环巴胺-KAAD组处理SMMC-7721细胞48 h,Caspase-3蛋白相对表达量分别为(0.36±0.05)、(0.67 ±0.10)、(0.89±0.11);bcl-2蛋白相对表达量分别为(0.61±0.10)、(0.48±0.05)、(0.24±0.08);CyclinD1蛋白分别为(1.41±0.21)、(0.71±0.08)、(0.53±0.11);p21蛋白分别为(0.49±0.08)、(0.97±0.09)、(1.55±0.13);Gli1、Shh的mRNA和蛋白表达均有不同程度的下降.Annexin V-FITC双染流式细胞术显示柠檬酸镧组和环巴胺-KAAD组增加SMMC-7721细胞凋亡率.结论 柠檬酸镧能抑制SMMC-7721细胞增殖、促进其凋亡.其机制可能和柠檬酸镧抑制Hh信号通路相关.

abstractsObjective To investigate the effect and the possible mechanism of Lanthanum Citrate on the proliferation and apoptosis of human hepatocellular carcinoma cell line SMMC-7721 through the Hedgehog signaling pathway.Methods Hhsignaling pathway inhibitor KAAD-cyclopamine and Lanthanum Citrate in different concentrations were used to treat human hepatocellular carcinoma cell line SMMC-7721.Cell proliferation was detected by MTT method.The apoptosis rate was detected by Annexin V/PI flow cytometry.The expressions of the regulative genes,such as CyclinD1,p21,Caspase-3,Bcl-2,Gli1,Shh were detected by Western blot.The mRNA expressions of Gli1 and Shh were detected by q-PCR.Results In control group,SMMC-7721 cells were treated with 0.1 mmol/L Lanthanum Citrate group,and 15 μnol/L KAAD-cyclopamine for 48 h,and expression of CyclinD1 protein in each group was (1.41 ± 0.21),(0.71 ±0.08) and (0.53 ±0.11);p21 protein was (0.49 ±0.08),(0.97 ±0.09) and (1.55 ±0.13);Caspase-3 protein was (0.36 ± 0.05),(0.67 ± 0.10) and (0.89 ± 0.11);the relative expression of Bcl-2 protein in each group was (0.61 ± 0.10),(0.48 ± 0.05) and (0.24 ± 0.08).Gli1,Shh mRNA and Gli1,Shh protein expression were decreased.AnnexinV-FITC double staining showed Lanthanum Citrate group and KAAD-cyclopamine group had increased apoptosis rate of SMMC-7721 cells by flow cytometry.Conclusion Lanthanum Citrate inhibits the proliferation and promotes apoptosis in hepatocellular carcinoma SMMC-772 cells through suppressing hedgehog signaling pathway.