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. 2021 Mar 22;11(4):187. doi: 10.1007/s13205-021-02739-2

Long non-coding RNA LIFR-AS1 regulates the proliferation, migration and invasion of human thyroid cancer cells

Sha Li 1,#, Chen Wang 1,#, Yifang Lu 1, Weijuan Li 1,✉
PMCID: PMC7985231  PMID: 33927978

Abstract

The long non-coding RNA (lncRNA) LIFR-AS1 has been shown to be involved in the development of several human cancers. This study was designed to determine the expression profile and role of lncRNA-LIFR-AS1 in human thyroid cancer. The results showed significant (p < 0.05) upregulation of LncRNA-LIFR-AS1 in thyroid cancer tissues and cells. However, silencing of LncRNA-LIFR-AS1 inhibited the viability and proliferation of human thyroid cancer cells inducing G2/M cell cycle arrest. The G2/M phase cells increased from 8.56% in negative control (NC) to around 35.03% in si-LIFR-AS1. This was also found to be concomitant with the downregulation of cyclin B1 and CDK1 expressions. The thyroid cancer cells exhibited remarkably lower invasion and migration under transcriptional knockdown of lncRNA-LIFR-AS1 which was also associated with downregulation of MMP-2 and MMP-9 expression. Importantly, transcriptional silencing of lncRNA-LIFR-AS1 inhibited thyroid cancer tumorigenesis, in vivo. Collectively, the results suggest the tumor-promoting role of lncRNA-LIFR-AS1 in thyroid cancer and highlight its potential as therapeutic target.

Keywords: Thyroid cancer, Long non-coding RNAs, LncRNA-LIFR-AS1, Cell cycle arrest, Xenograft

Introduction

Thyroid cancer is considered as the most common type of human cancer affecting endocrine system (Roman et al. 2017). At the global level, thyroid cancer has a prevalence of 66 out of 1 million human population (Siegel et al. 2016). During recent years, an increase in the thyroid cancer cases, particularly among the females has been reported (Schmidt and Davies 2017). Reportedly, more than 60,000 cases of thyroid cancer are reported in China annually (Chen et al. 2015). Although the exact cause of thyroid cancer is not known, a combination of genetic and environmental factors has been shown to be involved in its development and progression (Brent 2010). The therapeutic measures currently used against the thyroid cancer include surgical resection, hormone therapy and radio or chemotherapy. Nonetheless, they exhibit limited success at advanced stages. Recent years have witnessed a shift in understanding the therapeutic management of human cancers and researchers have proposed the investigation and therapeutic targeting of molecular pathways regulating the cancer progression (Jin et al. 2016). Thorough understanding of various molecular events governing the development and subsequent progression of thyroid cancer is lacking and the exploration of same is crucial to curb this disease in more effective and efficient fashion. There is concrete evidence regarding the involvement of long non-coding RNAs (lncRNAs) in human cancer (Zhang et al. 2016; Huarte 2015; Chen et al. 2017; Yang et al. 2018). The lncRNAS are seen with a promise to prove helpful in prognosis and molecular therapy of human cancers including the thyroid cancer (Zhang and Tang 2018; Gao and Wei 2017). LncRNA leukemia inhibitory factor receptor anti-sense RNA 1 (lncRNA-LIFR-AS1), one of the recently identified long non-coding RNA which is transcribed in an anti-sense fashion and has been shown to exhibit dysregulation in human cancers like breast cancer and colorectal cancer (Xu et al. 2019; Liu et al. 2018). Although lncRNA-LIFR-AS1 has been identified to be one of the 352 lncRNAs differentially expressed in thyroid cancer, its functional role has not been worked out in thyroid cancer (Liang et al. 2018). In the present study, we confirmed that thyroid cancer is linked with considerable upregulation of lncRNA-LIR-AS and is involved in the proliferation and tumorigenesis of thyroid cancer. The present study signifies the prognostic role and therapeutic applicability of lncRNA-LIFR-AS1 in human thyroid cancer.

Materials and methods

Clinical tissues

All the thyroid cancer tissues and matching normal adjacent clinical specimens were collected from 29 patients who underwent surgery at TangShan GongRen Hospital, TangShan, Hebei, China. Patients were informed in advance and written consents were obtained from them prior to tissue collection. The Institutional Ethics committee gave approval for this study. Furthermore, the standard guidelines passed by the Declaration of Helsinki were followed in strict sense for the conduct of various study procedures. The study was approved by the research ethics committee of TangShan GongRen Hospital TangShan, Hebei, China under approval number 62HT/GGH/2020.

Cell lines and culture conditions

Four different human thyroid cancer cell lines (MDA-T32, TT, K1 and MDA-T68) and normal human thyrocytes were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cell lines were cultured in RPMI-1640 medium (Gibco, Rockville, MD, USA) supplemented with 10% fetal bovine serum (FBS) (HyClone, Logan, UT), penicillin (100 U/mL) and streptomycin (100 mg/mL). The cells were maintained in an incubator with 5% CO2 at 37 °C using high humidity.

Cell transfection

The small interfering RNAs (siRNAs) against lncRNA-LIFR-AS1 (si-LIFR-AS1) and the non-specific negative control (si-NC) siRNAs were designed by Gene Pharma (Shanghai, China). To carry out the overexpression of lncRNA-LIFR-AS1 in thyroid cancer cells, its sequence was amplified and cloned into pcDNA3.1 (pcDNA-LIFR-AS1) overexpression plasmid (Invitrogen) while as the empty pcDNA3.1 vector was used as negative control. The transfection of thyroid cancer cells was performed using Lipofectamine2000 (Invitrogen) as per the manufacturer’s guidelines.

RNA isolation and RT-PCR

Total RNA from thyroid cancer tissues and cell lines along with the corresponding normal tissues and cells was extracted with the help the TRIzol reagent (Invitrogen) following the manufacturer protocol. Next, the RNA was reverse transcribed to synthesize the complementary DNA (cDNA) using PrimeScript™ RT Reagent kit (Invitrogen). For the expression analysis of lncRNA-LIFR-AS1, qRT‐PCR was performed on CFX96 quantitative PCR system (Bio‐Rad, Hercules, CA, USA) using SYBR Green Premix Ex Taq II (TaKaRa, Dalian, China). The 20 µL reaction mixture was composed of containing 1.5 mM MgCl2, 2.5 units Taq DNA Polymerase, 200 μM dNTP, 0.2 μM of each primer and 0.5 µg cDNA. The cycling conditions were as follows: 95˚C for 20 s, followed by 40 cycles of 95˚C for 15 s, and 57 °C for 1 min. Human GADPH was used as internal reference gene and 2−ddCt method was used for quantifying the relative transcript levels. The sequences of the primers used in Lnc-LIFRAS1 forward: 5′-AAGTTTCAGGCTCCTGACAGC-3′; Lnc-HANR-AS1 reverse: 5′-TTCGCCCACGTTCTTCTCGC-3′, GAPDH forward: 5′-AGAAGGCTGGGGCTCATTTG-3′; GAPDH reverse: 5′-AGGGGCCATCCACAGTCTTC-3′.

Cell viability assay

The MDA-T32 and MDA-T68 cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Briefly, the stably transfected cancer cells were added into 96-well plates with 4 × 104 cells/well. After being cultured for 0, 12, 24, 36, 48, 72 and 96 h, the medium was replaced, and the cells were subjected to treatment with MTT and incubated for another 2.5 h at 37 °C. Thereafter each well was supplemented with 150 μL dimethyl sulfoxide (DMSO) to dissolve the formazan crystals. Finally, the absorbance measurements at 570 nm were made using a spectrophotometer.

Colony formation assay

The stably transfected MDA-T32 and MDA-T68 thyroid cancer cells, at an initial density of 5000 cells per well were transferred into the six-well plates. The cells were grown at 37 °C using complete medium for a period of 2 weeks at which the colonies formed were ethanol fixed, stained with 0.2% crystal violet and photographed.

5-Ethynyl-2′-deoxyuridine (EdU) assay

The suspension of MDA-T32 and MDA-T68 transfected cancer cells carrying 104 cells was added into 24-well plate and cells were cultured for 24 h at 37 °C. Afterwards, each well was inoculated with BrdU labeling solution (Beyotime, Shanghai, China) and incubation of cancer cells was prolonged 8 h. Following, the media were removed and replaced with 70% ethanol and 2 mol/l HCl to allow fixation of cells and DNA denaturation, respectively. The cells were then washed thrice with phosphate‐buffered saline (PBS) and mixed with anti‐BrdU antibody (Beyotime) followed by their overnight incubation at 4 °C. The cells were then incubated with 4′,6‐diamidino‐2‐phenylindole (DAPI) solution (Beyotime) for 45 min for nuclei staining. Finally, after removal of staining solution, the cells were examined under a fluorescence microscope and photographed.

Cell cycle analysis

Following their stable transfection, the transfected MDA-T32 and MDA-T68 cancer cells were centrifuged and washed with cold PBS buffer twice. The cells fixed with 70% ethanol for 1.5 h and subsequently stained with Propidium iodide (PI) solution (Beyotime, Suzhou, China) for 15 min at room temperature. The PI-stained cells were subjected to cell cycle analysis with a flow cytometer (FACSCanto™ II, BD Biosciences).

Western blotting

The lysates of total proteins were obtained by digesting the radioimmunoprecipitation assay lysis buffer (RIPA; Beyotime, China). Proteins were quantified using bicinchoninic acid protein assay (Pierce, Appleton). Exactly, 40 µg of proteins were resolved using 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis following which they were transferred to polyvinylidene fluoride (PVDF) membranes (Millipore). 5% non-fat milk was used to block the membranes which were then incubated with specific primary antibodies overnight at 4 °C The membranes were subsequently exposed to a horseradish peroxidase‐conjugated secondary antibody for 2 h at room temperature. Finally, the protein bands were detected using an EasyBlot ECL detection system (Sangon, China). β-actin was used as a reference protein.

Cell invasion assays

Transwell chambers (8 μm pore size; Corning, Beijing, China) fitted with Matrigel (BD Biosciences, Franklin Lakes, NJ, USA) were employed for the investigation of cancer cell invasion. The transfected cells were firstly trypsinised and plated at a density of 5 × 105 into the upper compartment, while the lower compartment received only the culture medium. The cells were incubated at 37 °C for 24 h. Afterwards, the cells which failed to pass through the membrane were swabbed away while those which invaded the lower chamber were fixed using 4% paraformaldehyde and stained with 0.2% crystal violet. Finally, the cells were PBS washed and examined under an inverted microscope.

Cell migration assay

The migration of MDA-T32 and MDA-T68 cancer cells was analyzed using a scratch-heal assay. In brief, 5 × 104 cells were seeded into a 12-well plate. The cell surface was scratched with pipette-tip after cell attachment. The scratches were photographed at 0 and 24 h under an inverted microscope (Olympus, Japan) and their breadth was compared and analyzed using ImageJ v1.52.

In vivo tumorigenesis study

Nude male mice (7 weeks old) were divided into two groups randomly. Each animal was given an injection carrying 5 × 106 MDA-T32 thyroid cancer cells stably transfected with si-LIFR-AS1 or si-NC, intra-peritoneally. Following tumor induction, the tumor volume was determined after every 3 days with the help of a formula: tumor volume = 1/2[larger diameter × (smaller diameter)2]. The mice were euthanized after 3 weeks, and xenograft tumors were excised. The tumors were analyzed for relative size and their average weight was measured.

Statistical analysis

The study was performed using three replicates for each experiment and final values are representative of mean ± standard deviation (SD). The SPSS 13.0 (SPSS Inc.) software was used to perform the statistical analyses. Student’s t test and one-way ANNOVA were performed for analyzing the statistical differences. The p < 0.05 was considered to represent a statistically significant difference.

Results

LncRNA-LIFR-AS1 is upregulated in thyroid cancer

To gain insights into the probable regulatory role of lncRNA-LIFR-AS1 in thyroid cancer, the transcript levels of latter were investigated in thyroid cancer tissue and the normal matched human tissues. The cancer tissues exhibited significantly higher (p < 0.05) transcript levels of lncRNA-LIFR-AS1 in comparison to the normal tissues (Fig. 1a). The expression analysis was extended to different thyroid cancer cell lines (MDA-T32, TT, K1 and MDA-T68) using normal human thyrocytes as reference. The qRT-PCR study confirmed that the expression of lncRNA-LIFR-AS1 was significantly higher (p < 0.05) in all the four cancer cell lines (Fig. 1b). The MDA-T32 and MDA-T68 showed highest lncRNA-LIFR-AS1 expression among all the cancer cell lines. Collectively, these results are suggestive of lncRNA-LIFR-AS1 upregulation in thyroid cancer.

Fig. 1.

Fig. 1

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Knockdown of lncRNA-LIFR-AS1 inhibits proliferation of thyroid cancer cells. a Relative expression of lncRNA-LIFR-AS1 in thyroid cancer and matched normal human tissues (b) expression profiling of lncRNA-LIFR-AS1 from thyroid cancer cell lines (MDA-T32, TT, K1 and MDA-T68) and normal human thyrocytes (c) qRT-PCR based confirmation of lncRNA-LIFR-AS1 knockdown in MDA-T32 and MDA-T68 thyroid cancer cells with reference to corresponding si-NC control transfected cells (d) analysis of proliferation rate of MDA-T32 and MDA-T68 cancer cells transfected with si-LIFR-AS1 with reference to corresponding si-NC transfected control cells using MTT assay at different culture durations (e) analysis of colony formation by MDA-T32 and MDA-T68 cancer cells transfected with si-LIFR-AS1 with reference to corresponding si-NC transfected control cells (f) examination of proliferative viability of MDA-T32 cancer cells transfected with si-LIFR-AS1 or si-NC by EdU assay (g) examination of proliferative viability of MDA-T68 cancer cells transfected with si-LIFR-AS1 or si-NC by EdU assay. Data represents mean ± SD of three biological replicates (*p < 0.05)

LncRNA-LIFR-AS1 knockdown inhibits proliferation of thyroid cancer cells

To understand the role of lncRNA-LIFR-AS1 in regulating the growth and proliferation of thyroid cancer cells, MDA-T32 and MDA-T68 cells were transfected with small interfering RNAs designed against lncRNA-LIFR-AS1 (si-LIFR-AS1). Knockdown of lncRNA-LIFR-AS1 in MDA-T32 and MDA-T68 was confirmed by qRT-PCR taking corresponding silencing control (si-NC) transfected cells as negative control (Fig. 1c). The proliferation of si-LIFR-AS1 transfected MDA-T32 and MDA-T68 cancer cells was studied with reference to corresponding negative control cells with the help of MTT assay after 0, 12, 24, 48 or 96 h incubation at 37 °C. Both MDA-T32 and MDA-T68 cancer cells transfected with si-LIFR-AS1 proliferated at significantly lower (p < 0.05) rates than the negative control cells at the indicated culture durations (Fig. 1d). Again, both si-LIFR-AS1 transfected cancer cell lines showed significantly (p < 0.05) lower colony formation than the corresponding negative control cells (Fig. 1e). Furthermore, the EdU assay indicated that MDA-T32 cancer cells showed markedly lower proliferative viability when transfected with si-LIFR-AS1 (Fig. 1f). The proliferative viability of MDA-T68 was also affected in similar fashion under lncRNA-LIFR-AS1 knockdown (Fig. 1g). The results are thus conclusive of role of lncRNA-LIFR-AS1 in the development of thyroid cancer.

Overexpression of lncRNA-LIFR-AS1 enhances proliferation of thyroid cancer cells

Whether upregulation of lncRNA-LIFR-AS1 in thyroid cancer has any effect on cancer cell proliferation, lncRNA-LIFR-AS1 was first overexpressed in MDA-T32 cancer cells by transfecting them with pcDNA-LIFR-AS1 (overexpression construct) while empty vector transfected cells were used as reference (Fig. 2a). MTT assay was again performed to analyze the proliferation of lncRNA-LIFR-AS1 overexpressing cancer cells with reference to respective negative control cells. At different culture durations, the lncRNA-LIFR-AS1 overexpressing MDA-T32 exhibited significantly (p < 0.05) higher proliferation rate when compared to that of the negative control cells (Fig. 2b). Also, the colony formation from MDA-T32 cancer cells was remarkably increased under lncRNA-LIFR-AS1 overexpression (Fig. 2c). The results suggest that upregulation of lncRNA-LIFR-AS1 enhances the proliferation of thyroid cancer cells.

Fig. 2.

Fig. 2

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LncRNA-LIFR-AS1 overexpression enhanced thyroid cancer cell proliferation. a Overexpression of lncRNA-LIFR-AS1 in MDA-T32 cancer cells, control vector transfected cells served as negative control (b) MTT assay bases analysis of proliferation of MDA-T32 cancer cells transfected with overexpression vector construct of lncRNA-LIFR-AS1 (pcDNA-LIFR-AS1) or pcDNA3.1 empty vector (c) colony formation assay of MDA-T32 cancer cells transfected with overexpression vector construct of lncRNA-LIFR-AS1 (pcDNA-LIFR-AS1) or pcDNA3.1 empty vector. Data represents mean ± SD of three biological replicates (*p < 0.05)

LncRNA-LIFR-AS1 silencing induces G2/M cycle arrest in thyroid cancer cells

To unveil the possible mechanism underlying the proliferative decline of thyroid cancer cells under lncRNA-LIFR-AS1 knockdown, the flow cytometry was used to study the cell cycle distribution of si-NC or si-LIFR-AS1 MDA-T32 cancer cells. Interestingly, the percentage of MDA-T32 at G2/M phase was found to be 35.03% under lncRNA-LIFR-AS1 silencing, while it was only 8.56% for si-NC transfected cancer cells (Fig. 3a, b). Moreover, the expression levels of cell cycle regulatory proteins, Cyclin B1 and Cdk1 were considerably lower in si-LIFR-AS1 transfected MDA-T32 cancer cells in comparison to negative control cells (Fig. 3c). Taken together, the results are indicate that silencing of lncRNA-LIFR-AS1 induced G2/M cycle arrest in thyroid cancer cells.

Fig. 3.

Fig. 3

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LncRNA-LIFR-AS1 silencing induced G2/M cell cycle arrest in thyroid cancer cells. a Assessment of cell cycle phase distribution of si-NC control transfected MDA-T32 cancer cells by flow cytometry (b) assessment of cell cycle phase distribution of si-LIFR-AS1 transfected MDA-T32 cancer cells by flow cytometry (c) western blotting of Cyclin B1 and Cdk-1 cell cycle regulatory proteins from MDA-T32 cancer cells transfected with si-LIFR-AS1 or si-NC. Three replicates were used for performing each experiment

LncRNA-LIFR-AS1 silencing in thyroid cancer cells inhibits their migration and invasion

If lncRNA-LIFR-AS1 regulates the metastatic behavior of thyroid cancer cells, the invasion of MDA-T32 thyroid cancer cells transfected with si-LIFR-AS1 or si-NC was assessed using transwell chamber assay. The thyroid cancer cells showed considerably lower invasion under lncRNA-LIFR-AS1 silencing (Fig. 4a). The scratch-heal assay also showed that the migration of thyroid cancer cells declined considerably under lncRNA-LIFR-AS1 transcriptional knockdown (Fig. 4b). Besides, transfection of MDA-T32 cancer cells with si-LIFR-AS1 inhibited the expression of MMP2 and MMP9 proteins, markedly (Fig. 4c). Together, the results reveal that knockdown of lncRNA-LIFR-AS1 in thyroid cancer cells significantly reduces their invasion and migration, in vitro.

Fig. 4.

Fig. 4

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Silencing of lncRNA-LIFR-AS1 inhibits invasion and migration of thyroid cancer cells. a Transwell invasion assay of MDA-T32 cancer cells transfected with si-LIFR-AS1 or si-NC (b) scratch-heal migration assay of MDA-T32 cancer cells transfected with si-LIFR-AS1 or si-NC (c) western blotting of MMP-2 and MMP-9 proteins from MDA-T32 cancer cells transfected with si-LIFR-AS1 or si-NC. Three replicates were used for performing each experiment

Silencing of LncRNA-LIFR-AS1 inhibits in vivo tumorogenesis of thyroid cancer

Xenograft tumors were obtained by inducing the mice tumorogenesis through intra-peritoneal injections with MDA-T32 thyroid cancer cells. The MDA-T32 cancer cells used for induction of in vivo tumorogenesis were pre-transfected with si-LIFR-AS1 or si-NC. The mice tumors were excised 21 days after onset of mice tumorigenesis, and their relative sizes were compared. The volume of the si-LIFR-AS1 tumors was significantly (p < 0.05) lower size than those of si-NC tumors (Fig. 5a). Additionally, si-lncRNA-LIFR-AS1 tumors showed significantly (p < 0.05) lower tumor size as compared to the si-NC tumors (Fig. 5b). Collectively, the results suggest that lncRNA promotes the growth of thyroid cancer and its silencing inhibits the development of thyroid cancer.

Fig. 5.

Fig. 5

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Transcriptional knockdown of lncRNA-LIFR-AS1 inhibits in vivo thyroid tumorigenesis. a Analysis of relative xenograft tumor size from animal models inducted for tumorigenesis with MDA-T32 cancer cells transfected with si-LIFR-AS1 or si-NC (b) comparison of xenograft tumor size from animal models at different study stages inducted for tumorigenesis with MDA-T32 cancer cells transfected with si-LIFR-AS1 or si-NC (c) analysis of average xenograft tumor weight from animal models inducted for tumorigenesis with MDA-T32 cancer cells transfected with si-LIFR-AS1 or si-NC. Data represents mean ± SD of three biological replicates (*p < 0.05)

Discussion

Despite recent advancement, human cancer is still unquestionably one of the devastating health disorders across the globe (Hanahan 2014). Studies are being directed to identify potent therapeutic agents for the management of human cancers. LncRNAs have come into limelight for their crucial regulatory role in controlling the growth and progression of human cancers (Zhang et al. 2016; Huarte 2015; Chen et al. 2017; Yang et al. 2018). Thyroid cancer is associated with the dysregulation of number of lncRNAs (Liang et al. 2018; Lu et al. 2018). Nonetheless, the regulatory function of a novel lncRNA, LIFR-AS1 in thyroid cancer is still elusive. Several human cancers have been shown to be coupled with dysregulation of lncRNA-LIFR-(Xu et al. 2019; Liu et al. 2018; Wang et al. 2016). Consistently, in the present study, the thyroid cancer tissue and cell lines were revealed to express significantly higher transcript levels of lncRNA-LIFR-AS1 suggestive of a probable link with the onset and progression of thyroid cancer. The finding further points towards the probable utility of lncRNA-LIFR-AS1 in thyroid cancer prognosis which is in agreement with the findings of a previous study wherein the upregulation of lncRNA-LIFR-AS1 was found to be associated with the poor prognosis of gastric cancer (Wang et al. 2020a). As per the current study results, the knockdown of lnRNA-LIFR-AS1 inhibits the proliferation of thyroid cancer cells. On the other hand, overexpression of lncRNA-LIFR-AS1 in thyroid cancer cells positively affected their proliferation in vitro disclosing its tumor-promoting role. However, lncRNA-LIFR-AS1 acts as tumor suppressor in glioma and breast cancer signifying its dual role in human cancer pathogenesis (Xu et al. 2019; Ding et al. 2021). Silencing of lncRNA-LIFR-AS1 inducted thyroid cancer cells with G2/M cell cycle arrest and modulated the expression of cell cycle regulatory proteins, Cyclin B1 and Cdk1. The activation of latter proteins is required for the mitotic entry of dividing cells in eukaryotes (Jackman et al. 2003). Studies have shown that inhibition of Cyclin B1 or Cdk1 induces cell cycle arrest in cancer cells (Singh et al. 2017) which is consistent with the results of the present study. Several lncRNAs have been shown to regulate the migration and invasion of cancer cells (Mao et al. 2017). Consistently, we also examined the effects of lncRNA-LIFR-AS1 on the migration and invasion of the thyroid cancer cells. Interestingly, it was found that silencing of lncRNA-LIFR-AS1 suppressed the migration and invasion of the thyroid cancer cells. This is in confirmation with the results of a previous study wherein lncRNA-LIFR-AS1 was shown to regulate the migration and invasion of lung cancer cells (Wang et al. 2020b). The regulatory role of lncRNA-LIFR-AS1 in thyroid cancer metastasis was also evident from the inhibitory effects of lncRNA-LIFR-AS1 on the expression of matrix metalloproteins (MMPs) 2 and 9 which are pro-metastatic in function (Deryugina and Quigley 2006). Interestingly, the silencing of lncRNA-LIFR-AS1 significantly inhibited the tumorogenesis of mice xenograft tumors in vivo. The regulatory control of lncRNA-LIFR-AS1 on in vivo tumorogenesis has also been elucidated previously (Wang et al. 2020a).

Collectively, LncRNA-LIFR-AS1 exhibits significant upregulation in thyroid cancer. Knockdown of lncRNA-LIFR-AS1 inhibits growth of thyroid cancer cells via G2/M cell cycle arrest together with restraining their migration and invasion. In vivo thyroid cancer tumorogenesis was significantly minimized by lncRNA-LIFR-AS1 silencing. These results point towards the potential of lncRNA-LIFR-AS1 as biomarker and therapeutic target for the management of thyroid cancer.

Author contributions

Conceptualization: SL, CW and WL; methodology: SL, CW, YL and WL; formal analysis and investigation: SL, CW, YL and WL; writing—original draft preparation: SL, CW and WL; writing—review and editing critically for important intellectual content: WL; supervision: WL.

Funding

Not applicable.

Declarations

Ethical approval

A written informed consent was obtained from the patients before participation in the present study. The study was approved by the research ethics committee of TangShan GongRen Hospital TangShan, Hebei, China under approval number 62HT/GGH/2020.

Conflict of interest

All the authors declare that he has no conflict of interest.

Footnotes

Sha Li and Chen Wang contributed to this work equally.

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