柠檬酸镧对肝癌SMCC-7721细胞增殖凋亡的影响及其机制  

Effects and mechanism of Lanthanum Citrate on proliferation and apoptosis of human hepatocellular carcinoma cell line SMMC-7721

作　　者：陈斌[1] 戴正宽 李博飞 王春华[2] 

机构地区：[1]赣南医学院第一附属医院肝胆外科,江西赣州341000 [2]赣南医学院第一附属医院基础学院,江西赣州341000

出　　处：《中华肝胆外科杂志》2018年第1期49-53,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：江西省自然科学基金（20151BAB205044）;江西省教育厅科学技术研究项目（GJJ13687）

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AnnexinV／PI流式细胞术检测细胞凋亡率。采用蛋白印迹（Westernblot）检测CyclinDl、p21、Caspase-3、bcl-2、Glil、Shh表达。采用实时定量（q-PCR）检测Glil、Shh的表达。结果对照组、0．1mmol／L柠檬酸镧组、15μmol／L环巴胺-KAAD组处理SMMC-7721细胞48h，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）；CyclinDl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）；Glil、Shh的mRNA和蛋白表达均有不同程度的下降。AnnexinV-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。Objective To investigate the effect and the possible mechanism of Lanthanum Citrate on the proliferation and apoptosis of human hepatoeellular carcinoma cell line SMMC-7721 through the Hedgehog signaling pathway. Methods Hhsignaling pathway inhibitor KAAD-cyelopamine and Lanthanum Citrate in different concentrations were used to treat human hepatoeellular 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, Glil, Shh were detected by Western blot. The mRNA expressions of Glil and Shh were detected by q-PCR. Results In control group, SMMC-7721 cells were treated with 0.1 mmo]/L Lanthanum Citrate group, and 15 μmol/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）. Glil, Shh mRNA and Glil, 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.

关 键 词：肝细胞癌 信号通路 HEDGEHOG 柠檬酸镧 细胞增殖 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]