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Objective To investigate the effects of lncRNA SBF2-AS1 on the proliferation and invasion of hepatoma cells by regulating the miR-372-3p/CDK6 pathway. Methods Bel7402 and SK-hep1 cells were selected as research objects. The expression levels of SBF2-AS1, miR-372-3p, and CDK6 were up- or down-regulated according to different experimental stages, while the expression levels of miR-372-3p and CDK6 in cells were detected by real-time fluorescence quantitative PCR and Western blot. Dual luciferase reporter assay verified the targeting relationships between SBF2-AS1 and miR-372-3p as well as miR-372-3p and CDK6, respectively. CCK-8, colony formation assay, Transwell, cell cycle assay, and flow cytometry were used to analyze cell proliferation, colony formation, migration/invasion ability, cell cycle activity, and apoptosis. Results SBF2-AS1 was highly expressed in hepatocellular carcinoma cells (P<0.05). SBF2-AS1 knockdown resulted in decreased proliferation and invasion of Bel7402 and SK-hep1 cells (P<0.05). After miR-372-3p knockdown, the proliferation capacity and invasion number of Bel7402 cells were significantly increased. However, the above results were reversed after SBF2-AS1 knockdown (P<0.05). In addition, miR-372-3p targeted CDK6 and inhibited its expression, although over-expressing SFB2-AS1 could reverse the above results (P<0.05). Over-expressing CDK6 could reverse the inhibition of over-expressing miR-372-3p on the proliferation and invasion of Bel7402 cells. Conclusion LncRNA SBF2-AS1 can positively regulate the expression of CDK6 through miR-372-3p. It can also influence the distribution of cell cycle and affect the proliferation and invasion abilities of hepatocellular carcinoma cells.
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@# Objective: To investigate the effect of lncRNA SBF2-AS1 on epithelial-mesenchymal transition (EMT) of cervical cancer HeLa cell via regulating miR-140-5p/VEGFA (vascular endothelial growth factor A) axis. Methods: After cell culture and transfection, the cells were divided into 5 groups: NC group, miR-140-5p mimic group, miR-140-5p mimic+pcDNA-VEGFA group, si-lncRNA SBF2-AS1+pcDNA-VEGFA group and si-lncRNA SBF2-AS1+miR-140-5p mimic group. The expression level of lncRNA SBF2-AS1 in cervical cancer tissues and cell lines was detected by qPCR. The targeted relationship between lncRNA SBF2-AS1, miR-140-5p and VEGFA was confirmed by Dual luciferase reporter gene assay. The expression levels of VEGFA and EMT-related proteins N-cadherin, Vimentin and E-cadherin in HeLa cells were detected by WB. The invasion and migration of HeLa cells were detected by Transwell. Results: lncRNA SBF2-AS1 was highly expressed in cervical cancer tissues and cell lines (P<0.05 or P<0.01). Dual luciferase reporter gene assay confirmed that lncRNASBF2-AS1 targetedly combined with miR-140-5p and VEGFAwas a target gene of miR-140-5p (P< 0.05). Knockdown of lncRNA SBF2-AS1 inhibited invasion and migration as well as EMT of HeLa cells. Further experiment confirmed that lncRNA SBF2-AS1 up-regulated the expression level of VEGFA via miR-140-5p, thereby promoting invasion, migration and EMT of HeLa cells. Conclusion: lncRNASBF2-AS1 promotes EMT of HeLa cells via miR-140-5p/VEGFAaxis.
ABSTRACT
Objective To investigate the expression and clinical significance of long non-coding RNA SBF2-AS1 (SBF2 antisense RNA 1) in non-small cell lung cancer (NSCLC).Methods Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect SBF2-AS1 expressions in 114 cases of NSCLC tissues and the adjacent normal tissues to analyze its relationship with clinic-pathological characteristics,diagnostic value and prognosis in NSCLC.Results SBF2-AS1 expression was significantly higher in the NSCLC (4.336 ±0.032) compared with the adjacent normal tissues (1.256 ± 0.021),with a significant difference (t =3.594,P =0.005).The expression of SBF2-AS1 was related with tumor size (x2 =13.072,P =0.001),lymphatic metastasis (x2 =6.896,P =0.009),TNM stage (x2 =8.566,P =0.003),smoking history (x2 =8.769,P =0.003) and infiltration degree (x2 =17.852,P =0.001),but was not related with age (x2 =0.141,P =0.707),sex (x2 =0.036,P =0.850) and pathological type (x2 =1.267,P =0.260).The area under the receiver operating characteristic (ROC) curve was 0.853 (95 ％ CI:0.755-0.879,P =0.004).The sensitivity and specificity was 52.4％ and 87.8％,respectively.The difference betwen low SBF2-AS1 expression and high SBF2-AS1 expression groups was statistically significant in overall survival time (42.3 months vs.25.2 months,x2 =4.753,P =0.013).Conclusion The expression of SBF2-AS1 is upregulated in NSCLC and may be proved useful as a biomarker and diagnostic target for the treatment of patients with NSCLC.