학술논문
MicroRNA-200b inhibits pituitary tumor cell proliferation and invasion by targeting PKC[alpha]
Document Type
Report
Author
Source
Experimental and Therapeutic Medicine. August 2017, Vol. 14 Issue 2, p1706, 9 p.
Subject
Language
English
ISSN
1792-0981
Abstract
Introduction Pituitary adenomas generally account for 25% of intracranial tumors and can be divided into functioning and non-functioning pituitary adenomas. Non-functioning pituitary adenomas are not associated with sufficient hormone secretion, [...]
The present study aimed to investigate the expression of miR-200b and protein kinase C[alpha] (PKC[alpha]) in pituitary tumors and to determine whether miR-200b may inhibit proliferation and invasion of pituitary tumor cells. The regulation of PKC[alpha] expression was targeted in order to find novel targets for the treatment of pituitary tumors. In total, 53 pituitary tumor tissue samples were collected; these included 28 cases of invasive pituitary tumors and 25 cases of non-invasive tumors, in addition to 5 normal pituitaries. The expression level of miR-200b in the pituitary tumor tissue was detected by quantitative polymerase chain reaction (qPCR) and the expression of PKC[alpha] protein was detected by immunohistochemistry. A PKC[alpha] 3'untranslated region (UTR) luciferase vector was constructed and a dual luciferase reporter gene assay was employed in order to examine the effect of miR-200b on the PKC[alpha] 3'UTR luciferase activity. AtT-20 cells were transfected with miR-200b mimics, PKC[alpha] siRNA and miR-200b mimics + PKC[alpha], and the changes in cellular proliferation, invasion and apoptosis were observed via MTT, Transwell assay and flow cytometric analysis. Furthermore, PKC[alpha] mRNA expression was determined by qPCR, and Western blotting was performed to detect the expression of PKC[alpha] protein. miR-200b revealed downregulation in invasive pituitary tumor tissue, and the expression level was significantly down-regulated compared with normal and non-invasive pituitary tumor tissue (P Key words: pituitary tumor, miR-200b, protein kinase C-alpha, cell proliferation, invasion, apoptosis, AtT-20 cells, invasive pituitary
The present study aimed to investigate the expression of miR-200b and protein kinase C[alpha] (PKC[alpha]) in pituitary tumors and to determine whether miR-200b may inhibit proliferation and invasion of pituitary tumor cells. The regulation of PKC[alpha] expression was targeted in order to find novel targets for the treatment of pituitary tumors. In total, 53 pituitary tumor tissue samples were collected; these included 28 cases of invasive pituitary tumors and 25 cases of non-invasive tumors, in addition to 5 normal pituitaries. The expression level of miR-200b in the pituitary tumor tissue was detected by quantitative polymerase chain reaction (qPCR) and the expression of PKC[alpha] protein was detected by immunohistochemistry. A PKC[alpha] 3'untranslated region (UTR) luciferase vector was constructed and a dual luciferase reporter gene assay was employed in order to examine the effect of miR-200b on the PKC[alpha] 3'UTR luciferase activity. AtT-20 cells were transfected with miR-200b mimics, PKC[alpha] siRNA and miR-200b mimics + PKC[alpha], and the changes in cellular proliferation, invasion and apoptosis were observed via MTT, Transwell assay and flow cytometric analysis. Furthermore, PKC[alpha] mRNA expression was determined by qPCR, and Western blotting was performed to detect the expression of PKC[alpha] protein. miR-200b revealed downregulation in invasive pituitary tumor tissue, and the expression level was significantly down-regulated compared with normal and non-invasive pituitary tumor tissue (P Key words: pituitary tumor, miR-200b, protein kinase C-alpha, cell proliferation, invasion, apoptosis, AtT-20 cells, invasive pituitary