Knockdown of the c-Jun-N-terminal kinase expression by siRNA inhibits MCF-7 breast carcinoma cell line growth.
We have examined the effect of two small interference RNA against Jnk-1 and Jnk-2 in the breast cancer cell line MCF-7. The expression of the JNK-1 and JNK-2 is frequently elevated in breast cancer and is a frequent genetic abnormality in this malignancy. For a better understanding of its role in maintaining the malignant phenotype, we used small RNA interference (siRNA) directed against Jnk-1 or Jnk-2. We made control and Jnk-1 and Jnk-2 siRNA using vector plasmid, which was then transfected to reduce its expression in MCF-7 cells. We assessed the effects of JNK-1 or JNK-2 silencing on cell growth by western blot analysis, soft agar assay, cell proliferation assay, cell viability by MTT assay and caspase assay in vitro. Our data showed that siRNA against Jnk-1 or Jnk-2 markedly and durably reduced its expression in MCF-7 cells by up to 70%, decreased the growth rate of MCF-7 cells, inhibited colony formation in soft agar and significantly reduced cell growth in MCF-7 carcinoma culture cell line. We also found that depletion of JNK-1/2 in this manner promoted apoptosis of MCF-7 cells upon serum withdrawal. In addition, we found that MCF-7 cells did not exhibit any caspase-3 activity. In conclusion, we observed that JNK-1 and JNK-2 have a pivotal function in the development of breast cancer. Our data show that decreasing the JNK-1 or JNK-2 protein level in MCF-7 cells by siRNA could significantly inhibit MCF-7 cell growth in in vitro assay, and imply the therapeutic potential of siRNA on the treatment of breast cancer by targeting overexpression kinases such as JNK-1/2 and might be a potential therapeutic target for human breast cancer.
Parra
Eduardo
E
Biomedical Experimental Laboratory, Sciences Faculty, University of Tarapaca, Arica, Chile. eparra@uta.cl
Ferreira
Jorge
J
D017209:Apoptosis / Q000235:genetics; D001943:Breast Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D048708:Cell Growth Processes / Q000235:genetics; D045744:Cell Line, Tumor; D005260:Female; D055785:Gene Knockdown Techniques; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D048055:Mitogen-Activated Protein Kinase 8 / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics*; D048056:Mitogen-Activated Protein Kinase 9 / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D048056:Mitogen-Activated Protein Kinase 9; D048055:Mitogen-Activated Protein Kinase 8