(B,C) The effect of si-SNHG16 on cell proliferation was identified by MTT assay (Physique 3C,D). (for multiple groups). The statistical difference was defined as (Physique 2B,C). Simultaneously, transwell analysis discovered that the capacities of mobility and invasiveness were both reduced in SKNBE-2 and SK-N-SH cells (Physique 2D,E). In the mean time, the expression levels of E-cadherin, N-cadherin, and Vimentin were assessed using Western blot, the high expression of E-cadherin, and the low expression of N-cadherin and Vimentin showed the suppressive impact of SNHG16 silencing on epithelialCmesenchymal transition (EMT) (Physique 2FCI). These findings designed that knockdown of SNHG16 significantly constrained cell proliferation, migration, invasion, and EMT in NB cells. Open in a separate window Physique 2 SNHG16 deficiency hindered cell proliferation, migration, invasion, and EMT in NB cells(A) The knockdown efficiency of si-SNHG16 in SKNBE-2 and SK-N-SH cells was decided. (B,C) The effect of si-SNHG16 on cell proliferation was recognized by MTT assay (Physique 3C,D). At the same time, cell migration and invasion were analyzed in SKNBE-2 and SK-N-SH cells, and MTT analysis exhibited that the abilities of the mobility and invasiveness were evidently restrained (Physique 3E,F). In addition, the alteration of E-cadherin, N-cadherin, and Vimentin indicated that HNF4 silencing distinctly suppressed EMT in NB cells (Physique 3GCJ). The evidence displayed that HNF4 worked as an oncogenic role in SKNBE-2 and SK-N-SH cells. Open in a separate window Physique 3 HNF4 knockdown restrained cell proliferation, migration, invasion, and EMT (Physique 5GCJ). In brief, overexpression of HNF4 could abrogate the inhibiting effects of SNHG16 silencing on cell proliferation, migration, invasion, and EMT in NB cells. Open in a CEP-18770 (Delanzomib) separate window Physique 5 The impact of SNHG16 detetion on cell behaviors was regained by HNF4 up-regulation in NB cellsSKNBE-2 and SK-N-SH cells were transfected with si-NC, si-SNHG16, si-SNHG16+pcDNA, or si-SNHG16+pcDNA-HNF4, respectively, (A,B) and the protein level of HNF4 was estimated via Western blot. (C,D) The effects of si-SNHG16 and pcDNA-HNF4 on cell proliferation were measured. (E,F) The migrated cells or FLI1 invaded cells were counted and quantified by transwell CEP-18770 (Delanzomib) assay. (GCJ) Western blot assay was employed to determine the expression levels of E-cadherin, N-cadherin, and Vimentin. was our investigated object. First the stably transfected (lentivirus-mediated sh-SNHG16 or sh-NC) SKNBE-2 cells were injected into nude mice. After the killing of mice, we found that the xenograft tumor volumes and CEP-18770 (Delanzomib) weights were visibly decreased in sh-SNHG16 transfected group than that of sh-NC transfected group (Physique 6ACC). Then, the expression levels of SNHG16, miR-542-3p, and HNF4 were assessed by qRT-PCR, and the results displayed that this levels of SNHG16 and HNF4 were strikingly down-regulated, but miR-542-3p level was notably induced in treatment group (Physique 6D). Simultaneously, the protein expression level of HNF4 was clearly reduced in lentivirus-mediated sh-SNHG16 group (Physique 6E). All the data exhibited that SHKG16 detetion led to the decrease in NB tumor growth em in vivo /em . CEP-18770 (Delanzomib) Open in a separate window Physique 6 Knockdown of SNHG16 could curb the tumor growth em in vivo /em (ACC) The tumor volume and weight were recorded and analyzed after mice were killed. (D) qRT-PCR was carried out to evaluate the levels of SNHG16, miR-542-3p, and HNF4 in xenograft tumors. (E) Western blot was conducted to examine the protein expression level of mature HNF4 in tumor tissues. em *P /em 0.05. SNHG16 and HNF4 regulated the development of NB via RAS/RAF/MEK/ERK signaling pathway Based on the above introductions, we explored whether the RAS/RAF/MEK/ERK signaling pathway went in for the tumorigenic effects of SNHG16 and HNF4. Then, si-NC, si-SNHG16, si-SNHG16+pcDNA, or si-SNHG16+pcDNA-HNF4 was transfected into SKNBE-2 and SK-N-SH cells, respectively. We observed that SNHG16 detection specifically decreased the level of RAS, p-RAF, p-MEK, and p-ERK in SKNBE-2 cells, while the repressive impact of SNHG16 silencing was abolished after co-transfection with pcDNA-HNF4 (Physique 7A,B). Comparable phenomenon occurred in SK-N-SH cells (Physique 7C,D). In summary, SNHG16/miR-542-3p/HNF4 axis regulated NB progression via the activation of RAS/RAF/MEK/ERK signaling pathway (Physique 8). Open in a separate window Physique 7 SNHG16 and HNF4 regulated the development of NB via RAS/RAF/MEK/ERK signaling pathwaySi-NC, si-SNHG16, si-SNHG16+pcDNA, or si-SNHG16+pcDNA-HNF4 was launched into CEP-18770 (Delanzomib) SKNBE-2 and SK-N-SH cells, respectively. (A,B) The level changes of RAS, p-RAF, p-MEK, and p-ERK in SKNBE-2 cells were identified via Western blot assay. (C,D) The protein expression level changes of RAS, p-RAF, p-MEK, and p-ERK were detected through Western blot assay in SK-N-SH cells. em *P /em 0.05. Open in a separate window Physique 8 SNHG16/miR-542-3p/HNF4 axis regulated NB progression via RAS/RAF/MEK/ERK signaling pathway Conversation In the study, we reported that SNHG16 was highly expressed in.