LncRNA HOXA-AS2 regulates microRNA-216a-5p to promote malignant progression of non-small cell lung cancer
Authors: T-J Cui
J-P Zheng
Y Zheng
Z Chen
Q Chen
G-S Lin
Y-M Dai
X Lin
Journal: European review for medical and pharmacological sciences (2019)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: lncRNA; HOXA-AS2; non-small cell lung cancer; NSCLC; microRNA-216a-5p
Affiliations: Department of Medical Oncology, Fujian Provincial Hospital, Fuzhou, China. .

Abstract:  Objective: Previous studies have shown that long non-coding RNA (lncRNA) HOXA-AS2 is a cancer-promoting gene. However, the role of HOXA-AS2 in non-small cell lung cancer (NSCLC) has not been reported. This study aims to investigate the expression characteristics of HOXA-AS2 in NSCLC and whether HOXA-AS2 can promote the malignant progression of NSCLC by regulating microRNA-216a-5p. Patients and methods: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to examine the HOXA-AS2 level in 40 pairs of NSCLC tumor tissue samples and adjacent ones. Then, the relationship between HOXA-AS2 expression and clinical indicators and prognosis of NSCLC was analyzed. Meanwhile, qRT-PCR further verified the expression level of HOXA-AS2 in NSCLC cell lines. Also, HOXA-AS2 knockdown and overexpression models were constructed using lentivirus in NSCLC cell lines, and the effects of HOXA-AS2 on the biological function of NSCLC cells were analyzed using the Cell Counting Kit-8 (CCK-8), transwell migration, and cell wound healing assays. Finally, Western blotting assay and cell recovery experiment were used to explore the regulatory mechanism of HOXA-AS2 and microRNA-216a-5p in NSCLC. Results: In this experiment, qRT-PCR results revealed that HOXA-AS2 level in NSCLC tumor tissue specimens was remarkably higher than that in adjacent tissues. Compared with those with low expression of HOXA-AS2, the patients with high expression had a higher incidence of distant metastases and a lower overall survival rate. The proliferative and metastasis abilities of the cells in the HOXA-AS2 overexpression group were remarkably increased when compared with the control group, while the opposite results were observed in HOXA-AS2 silence group. Subsequently, qRT-PCR verified that microRNA-216a-5p level was remarkably decreased in NSCLC tissues and negatively correlated with HOXA-AS2 expression. In addition, the result of the cell recovery experiment and Western blotting revealed that there might be a mutual regulation between HOXA-AS2 and microRNA-216a-5p, the two of which could jointly regulate the malignant progression of NSCLC. Conclusions: The results indicate that lncRNA HOXA-AS2 is upregulated in NSCLC and is remarkably associated with distant metastasis and poor prognosis of NSCLC patients. In addition, lncRNA HOXA-AS2 is found to be able to promote the malignant progression of NSCLC via regulating microRNA-216a-5p. 

Retracted: No

PubPeer comments:
 Figure 2D:  , 
 Quantitation of cell number and % wound healing (Fig 2C,D and Fig 5C,D) looks more similar than by chance. Two different assays resulting in the same outcome?  , 
 From left to right: - Fig 1 from [Zang et al (2020)] - Fig 1 from [Li et al (2020)] - **Fig 1, and Fig 4D, from [Cui et al (2019)] **.  HOXA-AS2 is highly expressed in NSCLC tissues and cell lines. A, qRT-PCR was used to detect the difference in expression of HOXA-AS2 in NSCLC tumor tissues and adjacent tissues. B, qRT-PCR was used to detect the expression level of HOXA-AS2 in NSCLC cell lines. C, The Kaplan-Meier survival curve of lung cancer patients based on HOXA-AS2 expression ...  The Kaplan-Meier survival curve of lung cancer patients based on miR-216a-5p expression . - Fig 1D from [Gao et al (2019)] .  It appears that **any** combination of a cancer type, and the undesirable half of a non-coding RNA dichotomy, results in 100% mortality at 75 months. More examples: *[left]* Figure 1C from [Wu et al (2020)] . *[right]* Fig 3E from [Wu et al (2019)] .  ---------------------------------------------------------- 1C and 4D again:  Perhaps I misunderstand the plots. It seems that if the 40 cases are dichotomised into high- and low-HOXA-AS2 expression, both half-groups hit a survival rate of 0 at 75 months. If they are dichotomised into high- and low- microRNA-216a-5p, 20% of the low-expression half-group are still alive. , 
 *[left]* Fig 4C from [Cui et al (2019)] .  qRT-PCR showed a significant negative correlation between HOXA-AS2 and miR216a-5p expression in breast cancer tissues . *[right]* Fig 3C from [Pan, Song Gao (2019)] .  MiR-605-3p is significantly negatively correlated with the expression level of EZH2 in prostate cancer tissues . More points in common than one would expect.  Here, an overlay of 20 correlation scatterplots from various papers (all based on the same spreadsheet), emphasising the points frequently occurring between them.  ,