Silencing Endothelin-3 Expression Attenuates the Malignant Behaviors of Human Melanoma Cells by Regulating SPARC Levels  

作　　者：安湘杰 李艳秋 屈晓莺 张晶 张凌云 王明 朱里 陈思远 陈宏翔 涂亚庭 周育文 黄长征 

机构地区：[1]Department of Dermatology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology [2]Department of Dermatology,Zhongshan Hospital [3]Department of Dermatology,No.457 Airforce Hospital [4]Department of Dermatology,Wuhan General Hospital of Guangzhou Command [5]Department of Dermatology and Skin Science,University of British Columbia

出　　处：《Journal of Huazhong University of Science and Technology(Medical Sciences)》2013年第4期581-586,共6页华中科技大学学报（医学英德文版）

基　　金：supported partially by the grants from the National Natural Science Foundation of China(No.30671891,and No.81072232);the Canadian Foundation for Innovation,and the Canadian Institutes of Health Research(No.CIHR #IMH-37565 and No.MOP-84300)

摘　　要：Summary： Endothelin-3 （ET-3） is aberrantly expressed in both metastatic melanoma tissues and cul tured melanoma cells. Our previous work showed that ET-3 could promote survival of metastatic mela noma cells via its altered expression. In this study, we investigated the mechanisms responsible for these gene-induced phenotypes in melanoma cells. An ET-3 gene sequence-specific shRNA vector pLVTHM-ET3-RNAi was constructed and transfected into human malignant melanoma cells A375 and MMRU, and the resultant molecular events and cellular changes were examined. As compared with the empty-vector group, cell proliferation was slowed down, and the growth inhibition rates were 38.9% in A375 cells and 38.4% in MMRU cells after transfection. In addition, cell invasion capability was also inhibited, with a reduction of 62.2% in A375 cells and 54.3% in MMRU cells. The percentage of apoptotic cells was found to increase. Meanwhile, in both cell lines, secreted protein acidic and rich in cy teine （SPARC） levels were down-regulated together with inhibition of its upstream signaling molecule, NF-kB. Thus, the current results suggested that down-regulated expression of ET3 attenuates the ma lignant behaviors of human melanoma ceils partially by decreasing the expression of SPARC and NF-kB.Summary： Endothelin-3 （ET-3） is aberrantly expressed in both metastatic melanoma tissues and cul tured melanoma cells. Our previous work showed that ET-3 could promote survival of metastatic mela noma cells via its altered expression. In this study, we investigated the mechanisms responsible for these gene-induced phenotypes in melanoma cells. An ET-3 gene sequence-specific shRNA vector pLVTHM-ET3-RNAi was constructed and transfected into human malignant melanoma cells A375 and MMRU, and the resultant molecular events and cellular changes were examined. As compared with the empty-vector group, cell proliferation was slowed down, and the growth inhibition rates were 38.9% in A375 cells and 38.4% in MMRU cells after transfection. In addition, cell invasion capability was also inhibited, with a reduction of 62.2% in A375 cells and 54.3% in MMRU cells. The percentage of apoptotic cells was found to increase. Meanwhile, in both cell lines, secreted protein acidic and rich in cy teine （SPARC） levels were down-regulated together with inhibition of its upstream signaling molecule, NF-kB. Thus, the current results suggested that down-regulated expression of ET3 attenuates the ma lignant behaviors of human melanoma ceils partially by decreasing the expression of SPARC and NF-kB.

关 键 词：MELANOMA short hairpin RNA endothelin-3 nuclear factor-kappa B secreted proteinacidic and rich in cysteine 

分 类 号：R739.5[医药卫生—肿瘤]