Apoptosis-inducing activity of the antimicrobial peptide cecropin of Musca domestica in human hepatocellular carcinoma cell line BEL-7402 and the possible mechanism  

作　　者：Xiaobao Jin Hanfang Mei Xiaobo Li Yan Ma Ai-hua Zeng Yan Wang Xuemei Lu Fujiang Chu Qiang Wu Jiayong Zhu 

机构地区：[1]School of Basic Courses, Guangdong Pharmaceutical University, Guangzhou 510006, China [2]Guangdong Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, Guangzhou 510006, China

出　　处：《Acta Biochimica et Biophysica Sinica》2010年第4期259-265,共7页生物化学与生物物理学报（英文版）

基　　金：Funding This work was supported by grants from the National Natural Science Foundation of China （no. 30671832）, the Key Science and Technology Foundation of Guangdong Province （no. 2003B31602）, and the Key Science and Technology Foundation of Guangzhou （no. 2005Z3-E0211）.

摘　　要：We studied the apoptosis-inducing properties of the anti- microbial peptide cecropin of Musca domestica in human hepatocellular carcinoma cell line BEL-7402 and its underlying mechanism. Proliferation inhibition of the human hepatocellular carcinoma BEL-7402 cells and the human normal liver cells were determined by the MTT assay, and the cell viability was determined by trypan blue dye exclusion assay. The apoptotic tumor cells treated with cecropin were examined by transmission elec- tron microscopy and terminal-deoxynucleotidyl transfer- ase mediated nick end labeling. The apoptosis rate was measured by flow eytometry （FCM） with PI/Annexin-V double staining. Western blot analysis and RT-PCR were used to determine the expression levels of proteins involved in apoptosis, such as Fas, Fas-L, caspase-8, and caspase-3. The experimental results showed that Musca domestica cecropin inhibited the proliferation of human hepatocellular carcinoma BEL-7402 cells in dose- dependent and time-dependent manners, without affecting the proliferation of normal liver cells. FCM showed that the cell apoptosis rates were 5.1 ＋ 0.11%, 8.1 ＋ 0.04%, and 10.9 ＋ 0.15% after the treating with 100 uM eecropin for 24, 48, and 72 h, respectively. The rates of apoptosis were 5.4 ＋ 0.14% and 8.0 ＋ 0.13% after the treating with 25 and 50 p.M cecropin for 72 h, respectively. Western blot analysis and RT-PCR showed that the apoptosis- related molecules including Fas, Fas-L, caspase-8 and caspase-3 were activated. This study showed that the anti- microbial peptide ceeropin-inducing apoptosis in human hepatoeellular carcinoma BEL-7402 cells, which might be associated with upregulation of Fas, Fas-L, and caspase-8 and caspase-3 and triggering extrinsic apoptotic pathway.We studied the apoptosis-inducing properties of the anti- microbial peptide cecropin of Musca domestica in human hepatocellular carcinoma cell line BEL-7402 and its underlying mechanism. Proliferation inhibition of the human hepatocellular carcinoma BEL-7402 cells and the human normal liver cells were determined by the MTT assay, and the cell viability was determined by trypan blue dye exclusion assay. The apoptotic tumor cells treated with cecropin were examined by transmission elec- tron microscopy and terminal-deoxynucleotidyl transfer- ase mediated nick end labeling. The apoptosis rate was measured by flow eytometry （FCM） with PI/Annexin-V double staining. Western blot analysis and RT-PCR were used to determine the expression levels of proteins involved in apoptosis, such as Fas, Fas-L, caspase-8, and caspase-3. The experimental results showed that Musca domestica cecropin inhibited the proliferation of human hepatocellular carcinoma BEL-7402 cells in dose- dependent and time-dependent manners, without affecting the proliferation of normal liver cells. FCM showed that the cell apoptosis rates were 5.1 ＋ 0.11%, 8.1 ＋ 0.04%, and 10.9 ＋ 0.15% after the treating with 100 uM eecropin for 24, 48, and 72 h, respectively. The rates of apoptosis were 5.4 ＋ 0.14% and 8.0 ＋ 0.13% after the treating with 25 and 50 p.M cecropin for 72 h, respectively. Western blot analysis and RT-PCR showed that the apoptosis- related molecules including Fas, Fas-L, caspase-8 and caspase-3 were activated. This study showed that the anti- microbial peptide ceeropin-inducing apoptosis in human hepatoeellular carcinoma BEL-7402 cells, which might be associated with upregulation of Fas, Fas-L, and caspase-8 and caspase-3 and triggering extrinsic apoptotic pathway.

关 键 词：Musca domestica antimicrobial peptide CECROPIN hepatocellular carcinoma APOPTOSIS 

分 类 号：Q26[生物学—细胞生物学] Q516