G-D Zhang1, Y Li, G-J Liao, H-W Qiu. 1. Department of Sport Medicine, Yantaishan Hospital, Yantai, China. qiuhongwei86@outlook.com.
Abstract
OBJECTIVE: Our research explored the possible biological function of long non-coding RNA (lncRNA) NKILA in the pathogenesis of osteosarcoma and its underlying mechanism. PATIENTS AND METHODS: NKILA expression in 60 cases of osteosarcoma and adjacent tissues was detected. The correlation between NKILA expression and clinical information was analyzed by Chi-square test. The overexpression plasmid or siRNA of NKILA were transfected into osteosarcoma cells by liposome. Cell proliferation was detected by cell counting kit-8 (CCK-8) assay. Transwell assay was used to check the migratory and invasive abilities. Western Blot was used to detect the expressions of nuclear factor-κB (NF-κB)-related proteins. In addition, we analyzed the cell invasion and migration after treatment of NF-κB inhibitor (JSH) to further verify whether NKILA can participate in the occurrence of osteosarcoma through the NF-κB / Snail signaling pathway. RESULTS: The expression level of NKILA in osteosarcoma tissues was significantly lower than that in adjacent tissues, and was related to tumor size, Enneking stage, and metastasis. After KNKS/NP cells were transfected with NKILA-siRNA, cell proliferation, invasion and migration were enhanced. Transfection of the NKILA overexpression plasmid in Saos2 cells reduced cell proliferation, invasion and migration. NKILA knockdown downregulated the expressions of p65 and E-cadherin, but strikingly increased Snail expression. The RNA binding protein co-immunoprecipitation experiments illustrated that p65 could bind to NKILA. Additionally, JSH was found to reverse the inhibitory effect of NKILA on cell migration and proliferation. CONCLUSIONS: NKILA was lowly expressed in osteosarcoma tissues. In addition, high expression of NKILA could suppress the migration and invasion of osteosarcoma cells by inhibiting the NF-κB/Snail signaling pathway.
OBJECTIVE: Our research explored the possible biological function of long non-coding RNA (lncRNA) NKILA in the pathogenesis of osteosarcoma and its underlying mechanism. PATIENTS AND METHODS: NKILA expression in 60 cases of osteosarcoma and adjacent tissues was detected. The correlation between NKILA expression and clinical information was analyzed by Chi-square test. The overexpression plasmid or siRNA of NKILA were transfected into osteosarcoma cells by liposome. Cell proliferation was detected by cell counting kit-8 (CCK-8) assay. Transwell assay was used to check the migratory and invasive abilities. Western Blot was used to detect the expressions of nuclear factor-κB (NF-κB)-related proteins. In addition, we analyzed the cell invasion and migration after treatment of NF-κB inhibitor (JSH) to further verify whether NKILA can participate in the occurrence of osteosarcoma through the NF-κB / Snail signaling pathway. RESULTS: The expression level of NKILA in osteosarcoma tissues was significantly lower than that in adjacent tissues, and was related to tumor size, Enneking stage, and metastasis. After KNKS/NP cells were transfected with NKILA-siRNA, cell proliferation, invasion and migration were enhanced. Transfection of the NKILA overexpression plasmid in Saos2 cells reduced cell proliferation, invasion and migration. NKILA knockdown downregulated the expressions of p65 and E-cadherin, but strikingly increased Snail expression. The RNA binding protein co-immunoprecipitation experiments illustrated that p65 could bind to NKILA. Additionally, JSH was found to reverse the inhibitory effect of NKILA on cell migration and proliferation. CONCLUSIONS:NKILA was lowly expressed in osteosarcoma tissues. In addition, high expression of NKILA could suppress the migration and invasion of osteosarcoma cells by inhibiting the NF-κB/Snail signaling pathway.