LncSNHG16 promotes proliferation and migration of osteosarcoma cells by targeting microRNA-146a-5p
Authors: J-W Yan
J Tang
Y Ni
S-N Zheng
J Yang
X Chen
D-W Ge
J-J Qiu
L Zhao
L Yang
Z-W Yin
Journal: European review for medical and pharmacological sciences (2019)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: lncSNHG16; osteosarcoma; microRNA-146a-5p; NOVA1; tumor stage
Affiliations: Department of Orthopedic Surgery, Nanjing First Hospital, Nanjing Medical University, Nanjing, China. .

Abstract:  Objective: The aim of this study was to elucidate the regulatory role of lncSNHG16 in the progression of osteosarcoma (OS) and its underlying mechanism. Materials and methods: Expressions of lncSNHG16, microRNA-146a-5p and NOVA1 in OS tissues and adjacent normal tissues were determined by quantitative Real-time polymerase chain reaction (qRT-PCR). Their expressions in OS cell lines were detected by qRT-PCR as well. We analyzed the relationship between lncSNHG16 expression and tumor stage, diagnosis and survival prognosis of OS patients, respectively. Cell counting kit-8 (CCK-8) and transwell experiments were conducted to explore proliferative and migratory changes of OS cells. Dual-luciferase reporter assay was used to verify the binding relationship of lncSNHG16 to microRNA-146a-5p, and microRNA-146a-5p to NOVA1. Finally, rescue experiments were performed to elucidate the regulatory effect of lncSNHG16 on the cellular behaviors of OS cells. Results: LncSNHG16 was highly expressed in OS tissues and cell lines. Its expression was positively correlated with the tumor stage of OS patients. Receiver operating characteristic (ROC) curves suggested that lncSNHG16 can be used as a clinical indicator to distinguish OS patients from healthy controls. Survival analysis indicated a negative correlation between lncSNHG16 expression and survival of OS patients. Overexpression of lncSNHG16 enhanced the proliferative and migratory potentials of OS cell lines 143B and MNNG/HOS. MicroRNA-146a-5p was predicted to be the target gene of lncSNHG16, which was lowly expressed in OS tissues and cell lines. Overexpression of lncSNHG16 downregulated the expression of microRNA-146a-5p in 143B and MNNG/HOS cells. Furthermore, we verified that lncSNHG16 could bind to microRNA-146a-5p. The promotive role of lncSNHG16 in proliferative and migratory potentials of OS cells was reversed by microRNA-146a-5p. Subsequently, NOVA1 was predicted to be the target gene of microRNA-146a-5p, and was further verified by dual-luciferase reporter gene assay. Correlation analysis showed that microRNA-146a-5p expression was negatively correlated with NOVA1 expression in OS. More importantly, NOVA1 reversed the inhibitory effect of microRNA-146a-5p on the proliferative and migratory capacities of 143B and MNNG/HOS cells. Conclusions: LncSNHG16 is highly expressed in OS tissues and cell lines, participating in the development of OS by downregulating microRNA-146a-5p to upregulate NOVA1 expression. 

Retracted: No

PubPeer comments:
  LncSNHG16 was highly expressed in OS tissues compared with adjacent normal tissues (Figure 1A). Furthermore, we found that advanced OS patients had a higher expression of lncSNHG16 than those early-stage OS patients, suggesting that lncSNHG16 expression may be related to tumor stage of OS (Figure 1B). ![file] Curiously, when 1B is rescaled to the same vertical axis as 1A, the extremes of the Normal Control distributions are different. The combined extremes of the Advanced and Early-Stage distributions do not match the Combined distribution. ![file] In fact the Advanced OS distribution in 1B has **the same median** as the **total OS** distribution in 1A, which makes no sense at all. , 
 #1 Dear Prof. Hoya Camphorifolia, thanks very much for your comments. We are sory about the delay of our reply, as I just found the email reminder today. After checking your comments, we immediately checked the original data. Actually, the samples used for Fig 1A and Fig 1B were different, from different patients. The purpose of Fig 1A was to explore whether SNHG16 in OS tissues was highly expressed compared with their adjecent control tissues (patients samples in Fig 1A were all diagnosed with OS). In Fig B, we used different patient samples to invetigate whether tumor stage affects SNHG16 expression. As you can see, we used normal group to distinguish it from control group. These normal tissues were from the patients who were diagnosed without OS via pathological examination. Here, we provided some of the samples data. We are so sorry that we did not make the detailed descriptions in the text or Figure legends. We will contact the editor to add the detailed description about the above samples selection. Thanks again for your valuable comments and suggestions. , 
 #1 ![file] Figure 1A ![file] Figure 1B , 
 #2 The use of tissue samples from **non**-osteosarcoma patients should probably have been mentioned in the paper somewhere. , 
 I agree with Hoya in #4, that the sudden appearance of a control group that was not mentioned in the paper is of concern. Even more concerning, what type of control patients were used to serve as non-osteosarcoma patients, assuming that the tissues taken for this study were bone or bone related? I assume there would not be a lot of healthy volunteers donating bone tissues for such a study. Also, what was the age range of either the patient group or the control group? Again, the paper does not provide any details, which is of concern since osteosarcoma affects predominantly children and adolescents (source: this paper). The paper does not mention anything about informed consent or approval by parents or guardians of minors. , 
 Could the authors also please check Figures 2E and 3H? Two panels appear to look very similar, although they represent different experiments. Marked with red boxes. ![file] ,