Cystatin E/M suppresses legumain activity and invasion of human melanoma.
Cystatin E/M suppresses legumain activity and invasion of human melanoma.
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DOI：
10.1186/1471-2407-10-17
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发表时间：
2010-01-15
期刊：
影响因子：
3.8
通讯作者：
Solberg R
中科院分区：
文献类型：
--
作者：
Briggs JJ;Haugen MH;Johansen HT;Riker AI;Abrahamson M;Fodstad Ø;Maelandsmo GM;Solberg R
半胱氨酸蛋白酶的高活性，如豆蛋白和组织蛋白，已被证明促进了各种类型的肿瘤的生长和侵袭。在乳腺癌中，最近的几项研究表明，半胱氨酸蛋白酶抑制剂Cystatin E/M的缺失会导致生长和转移的增加。尽管Cystatin E/M在皮肤中正常表达，但其在半胱氨酸蛋白酶调节和恶性黑色素瘤进展中的作用尚未被研究。使用了一组不同的非黑色素瘤和黑色素瘤细胞系。用免疫印迹和酶联免疫吸附试验检测细胞培养液中胱抑素E/M和C的表达。用免疫印迹法检测细胞裂解物中的豆蛋白、组织蛋白酶B和L，并用多肽底物分析它们的酶活性。将Cstatin E/M表达载体(PCST6)导入两株未检测到cystatin E/M分泌的黑色素瘤细胞系，通过Matrigel侵袭实验检测细胞的迁移和侵袭能力。在所有已建立的黑色素瘤细胞系的培养液中均未检测到半胱氨酸氨基转移酶E/M，而在5个原发黑色素瘤细胞系中的2个和6个转移性疾病患者的6个黑色素瘤细胞系中有2个检测到强免疫条带。在4株分泌半胱氨酸氨基转移酶E/M的黑色素瘤中，糖基化形式(17kD)占优势，非糖化形式(14kD)占优势。虽然在表达cystatin E/M的黑色素瘤中低水平表达cystatin E/M，但在分泌cystatin E/M的黑色素瘤细胞系中，cystatin C一般不表达或表达很低。将pCST6基因导入缺乏胱抑素E/M分泌的黑色素瘤细胞，其胞内豆蛋白活性受到明显抑制。相反，组织蛋白酶B的活性没有受到影响。此外，Transwell Matrigel实验表明，在cystatin E/M过表达的黑色素瘤细胞系中，侵袭力受到抑制。这些结果表明，胱抑素E/M水平调节豆蛋白活性，从而调节人类黑色素瘤细胞的侵袭潜能。
High activity of cysteine proteases such as legumain and the cathepsins have been shown to facilitate growth and invasion of a variety of tumor types. In breast cancer, several recent studies have indicated that loss of the cysteine protease inhibitor cystatin E/M leads to increased growth and metastasis. Although cystatin E/M is normally expressed in the skin, its role in cysteine protease regulation and progression of malignant melanoma has not been studied. A panel of various non-melanoma and melanoma cell lines was used. Cystatin E/M and C were analyzed in cell media by immunoblotting and ELISA. Legumain, cathepsin B and L were analyzed in cell lysates by immunoblotting and their enzymatic activities were analyzed by peptide substrates. Two melanoma cell lines lacking detectable secretion of cystatin E/M were transfected with a cystatin E/M expression plasmid (pCST6), and migration and invasiveness were studied by a Matrigel invasion assay. Cystatin E/M was undetectable in media from all established melanoma cell lines examined, whereas strong immunobands were detected in two of five primary melanoma lines and in two of six lines derived from patients with metastatic disease. Among the four melanoma lines secreting cystatin E/M, the glycosylated form (17 kD) was predominant compared to the non-glycosylated form (14 kD). Legumain, cathepsin B and L were expressed and active in most of the cell lines, although at low levels in the melanomas expressing cystatin E/M. In the melanoma lines where cystatin E/M was secreted, cystatin C was generally absent or expressed at a very low level. When melanoma cells lacking secretion of cystatin E/M were transfected with pCST6, their intracellular legumain activity was significantly inhibited. In contrast, cathepsin B activity was not affected. Furthermore, invasion was suppressed in cystatin E/M over-expressing melanoma cell lines as measured by the transwell Matrigel assay. These results suggest that the level of cystatin E/M regulates legumain activity and hence the invasive potential of human melanoma cells.
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影响因子：
3.2
作者：
GUESDON, JL;TERNYNCK, T;AVRAMEAS, S
通讯作者：
AVRAMEAS, S
影响因子：
11.2
作者：
Kim, Tae-You;Zhong, Sheng;Robertson, Keith D.
通讯作者：
Robertson, Keith D.
DOI：
10.1016/0305-0491(96)00025-9
发表时间：
1996-07-01
影响因子：
2.2
作者：
Hakansson, K;Huh, C;Abrahamson, M
通讯作者：
Abrahamson, M
影响因子：
8
作者：
Lakka, SS;Gondi, CS;Rao, JS
通讯作者：
Rao, JS
影响因子：
2.9
作者：
Klose, Anke;Zigrino, Paola;Hunzelmann, Nicolas
通讯作者：
Hunzelmann, Nicolas