J-R Zhang1, R-H Zhu, X-P Han. 1. Department of Physical Examination Center, Jinan Central Hospital affiliated to Shandong University, Jinan, Shandong, China. j627843774@sina.com.
Abstract
OBJECTIVE: Larynx carcinoma is a common head-neck malignant tumor. Recent investigations showed the involvement of microRNA (miR) in regulation of multiple tumors. miR-140-5p showed decreased expression in various cancers, but without knowledge regarding its expression in larynx carcinoma and effects on cell invasion and angiogenesis. MATERIALS AND METHODS: Real-time quantitative PCR was firstly employed to measure miR-140-5p expression in larynx carcinoma and controlled tumor adjacent tissues. In larynx cancer cell line, agomir or antagomir of miR-140-5p was applied to up-regulate or down-regulate miR-140-5p, respectively. Western blot was used to evaluate vascular endothelial growth factor A (VEGF-A) expression, and cell proliferation was modified by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H- tetrazolium bromide (MTT) approach. Transwell approach was used to measure cell invasion, and angiogenesis assay was used to detect the effect on angiogenesis. Luciferase report assay (LRA) measured targeting binding between miR-140-5p and VEGF-A. RESULTS: Comparing to tumor adjacent tissues, larynx carcinoma cells showed significantly decreased miR-140-5p expression. Agomir up-regulated miR-140-5p expression and weakened proliferation and invasion potency, and inhibited angiogenesis. Antagomir down-regulated miR-140-5p and presented the opposite results. Finally, LRA confirmed direct binding between miR-140-5p and VEGF-A. CONCLUSIONS: MiR-140-5p can target VEGF-A in larynx carcinoma cell line to inhibit cell invasion and angiogenesis. MiR-140-5p thus may work as the direct molecular target of larynx carcinoma.
OBJECTIVE:Larynx carcinoma is a common head-neck malignant tumor. Recent investigations showed the involvement of microRNA (miR) in regulation of multiple tumors. miR-140-5p showed decreased expression in various cancers, but without knowledge regarding its expression in larynx carcinoma and effects on cell invasion and angiogenesis. MATERIALS AND METHODS: Real-time quantitative PCR was firstly employed to measure miR-140-5p expression in larynx carcinoma and controlled tumor adjacent tissues. In larynx cancer cell line, agomir or antagomir of miR-140-5p was applied to up-regulate or down-regulate miR-140-5p, respectively. Western blot was used to evaluate vascular endothelial growth factor A (VEGF-A) expression, and cell proliferation was modified by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H- tetrazolium bromide (MTT) approach. Transwell approach was used to measure cell invasion, and angiogenesis assay was used to detect the effect on angiogenesis. Luciferase report assay (LRA) measured targeting binding between miR-140-5p and VEGF-A. RESULTS: Comparing to tumor adjacent tissues, larynx carcinoma cells showed significantly decreased miR-140-5p expression. Agomir up-regulated miR-140-5p expression and weakened proliferation and invasion potency, and inhibited angiogenesis. Antagomir down-regulated miR-140-5p and presented the opposite results. Finally, LRA confirmed direct binding between miR-140-5p and VEGF-A. CONCLUSIONS:MiR-140-5p can target VEGF-A in larynx carcinoma cell line to inhibit cell invasion and angiogenesis. MiR-140-5p thus may work as the direct molecular target of larynx carcinoma.