鲨鱼软骨制剂对人肝癌SMMC-7721细胞中Bcl-2和Caspase-3表达的影响  

Effect of SCP on the expression of Bcl-2 and Caspase-3 in SMMC-7721 cells of hepatocarcinoma

作　　者：李伟[1] 陈小义[2] 刘英富[2] 李威[2] 

机构地区：[1]解放军二五四医院消化科,天津300142 [2]武警后勤学院细胞生物与医学遗传学教研室,天津300309

出　　处：《武警后勤学院学报（医学版）》2017年第9期746-750,共5页Journal of Logistics University of PAP（Medical Sciences）

基　　金：武警部队后勤学院基金资助项目(WYB201116)

摘　　要：【目的】检测鲨鱼软骨制剂(shark cartilage preparation,SCP)对人肝癌SMMC-7721细胞的抑制效果及对凋亡调控蛋白Bcl-2和Caspase-3的影响。【方法】以不同浓度SCP加入体外培养的对人肝癌SMMC-7721细胞中,MTT法检测SCP对细胞生长抑制作用;Hoechst33342/PI双荧光染色法考察SCP对细胞凋亡的影响;采用流式细胞术比较不同处理后的对人肝癌SMMC-7721细胞凋亡水平;免疫细胞化学染色法检测不同组别细胞凋亡相关蛋白Bcl-2和Caspase-3蛋白的表达水平;Western blotting检测Bcl-2蛋白和切割型Caspase-3(Cleaved-Caspase-3)蛋白的表达。【结果】SCP明显抑制对人肝癌SMMC-7721细胞生长,且抑制作用呈浓度-作用时间依赖性(P<0.05),其24、48和72 h的IC50值分别为1.27、1.86和0.84 mg。1 mg/ml SCP处理细胞24 h,细胞出现核染色质凝集、凋亡小体等典型凋亡特征,随着处理时间延长,细胞凋亡比例明显增大并表现出时间依赖性(P<0.05);免疫细胞化学、Western blotting检测显示SCP诱导细胞凋亡过程中Bcl-2蛋白表达减弱,Caspase-3蛋白表达明显增强。【结论】SCP可以抑制对人肝癌SMMC-7721细胞的增殖及诱导细胞凋亡,其诱导凋亡可能与下调Bcl-2蛋白表达并上调Caspase-3蛋白的表达有关。[ Objective ]To explore the inhibitory effect of shark cartilage preparation （SCP） on human hepatocarcinoma cell line SMMC- 7721 and its effect on the expression of Bcl-2 and Caspase-3. [Methods] SCP in different concentrations was added into the media of SMMC-7721 cells euhured in vitro. The inhibitory effect of SCP was measured by MTT method. Cells morphologie changes were determined by Hoeehest33342/PI double fluorescent staining. The level of apoptosis was detected by flow cytometry. The expression levels of apoptosis-related proteins Bel-2 and Caspase-3 were measured with immunoeytoehemistry. The protein expression levels of Bel-2 and Cleaved-Caspase-3 were detected by Western blot. [ Results] SCP could obviously inhibit the growth of SMMC-7721 cells in a dose-and-time dependent manner （P〈0.05）. The IC50 values of SCP for SMMC-7721 cells at 24, 48, 72 h were 1.27 mg, 1.86 mg and 0.84 mg respectively. SMMC-7721 cells treated with 1 mg/ml SCP for 24 h showed some typical apoptosis characteristics such as nuclei ehromatin condensation and apoptotic body. The proportion of apoptosis cells strikingly increased in time-dependent manner （P〈 0.05）. The results of immunoeytochemistry and Western blotting showed that the level of Bel-2 protein in SMMC-7721 cells treatedwith SCP obviously decreased, while the level of caspase-3 significantly increased. [Conclusion] SCP can inhibit the proliferation of SMMC-7721 cells and induce the apoptosis, and its mechanism might be related to the down-regulation of Bcl-2 expression and up-regulation of Caspase-3 expression.

关 键 词：鲨鱼软骨制剂 人肝癌SMMC-7721细胞 细胞凋亡 

分 类 号：R735[医药卫生—肿瘤]