title: PKCα is involved in the progression of kidney carcinoma through regulating netrin-1/UNC5B signaling pathway.
abstract: With a special interest towards a better understanding of signal pathways, we attempted to discover a safer and more effective therapeutic strategy for kidney carcinoma. Recent studies had suggested a role mediated by PKCα for netrin-1 and its receptors in the initiation and progression of tumors. Real-time PCR and western blotting were used to determine the expression levels of netrin-1 and UNC5B. We made use of the agonist of PKCα (phorbol-12-myristate 13-acetate-PMA) and the inhibitor of PKCα (calphostin C) to treat renal cell carcinoma (RCC) cells, and MTT assays were used to measure cell proliferation. By immunofluorescence, we identified the localization of netrin-1 and UNC5B in RCC cell lines 769-P and ACHN. The expression of UNC5B in tumor tissues was significantly downregulated compared to the corresponding normal tissues in which netrin-1 was upregulated. In low grade tumors, UNC5B expression was more prominent while netrin-1 expression was the opposite when compared with high grade ones. Proliferation of ACHN cells was concentration dependent in the presence of PMA and calphostin C. Netrin-1 and UNC5B expressions were upregulated in cells treated with PMA while calphostin C reversed this upregulation. By immunofluorescence, we identified that netrin-1 was highly expressed in the nuclear but none of UNC5B. Our data highly suggested that PMA-induced upregulation and calphostin C-induced reversion of netrin-1 and UNC5B in kidney carcinoma were accompanied by the activation of the netrin-1/UNC5B pathways.
authors.affiliation: Department of Urology, The First Hospital of China Medical University, 155 Nanjing North Street, Heping District, Shenyang City, Liaoning Province, People's Republic of China, 110001. meinv060523@163.com
authors.forename: Bo
authors.initials: B
authors.lastname: Zhan
authors.forename: Chuize
authors.initials: C
authors.lastname: Kong
authors.forename: Kunfeng
authors.initials: K
authors.lastname: Guo
authors.forename: Zhe
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D016022:Case-Control Studies; D049109:Cell Proliferation / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007124:Immunoenzyme Techniques; D007680:Kidney Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D009281:Naphthalenes / Q000494:pharmacology; D060787:Neoplasm Grading; D009367:Neoplasm Staging; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism*; D000075386:Netrin Receptors; D000075388:Netrin-1; D010703:Phorbol Esters / Q000494:pharmacology; D051571:Protein Kinase C-alpha / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; C000617351:NTN1 protein, human; D009281:Naphthalenes; D009414:Nerve Growth Factors; D000075386:Netrin Receptors; D010703:Phorbol Esters; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D025521:Tumor Suppressor Proteins; C532445:UNC5B protein, human; D000075388:Netrin-1; C119222:phorbol-12-myristate; C496628:PRKCA protein, human; D051571:Protein Kinase C-alpha; C058819:calphostin C