Downregulation of MIAT reduces the proliferation and migratory and invasive abilities of retinoblastoma cells by sponging miR‑665 and regulating LASP1

  • Authors:
    • Xiabing Xu
    • Yadong Zhao
    • Gang Duan
    • Bo Du
  • View Affiliations

  • Published online on: September 22, 2021     https://doi.org/10.3892/etm.2021.10777
  • Article Number: 1342
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Long non‑coding RNAs (lncRNAs) can function as onco‑lncRNAs in several types of human cancer, including retinoblastoma (Rb). The present study investigated the potential role and regulatory mechanism of the lncRNA myocardial infarction‑associated transcript (MIAT) in Rb. To do so, the expression levels of MIAT, microRNA (miR)‑665, and LIM and SH3 protein 1 (LASP1) in Rb tissues from patients or Rb cells were analysed using reverse transcription quantitative PCR. The interactions between miR‑665 and MIAT/LASP1 were confirmed by the dual‑luciferase reporter assay. MTT, Transwell (to assess migration and invasion) and western blotting assays were used to explore the functions of the MIAT/miR‑665/LASP1 axis on Rb progression in vitro. The results of the present study indicated that MIAT targeted miR‑665. In Rb tissues and cell lines, high expression of MIAT was observed, whereas miR‑665 was downregulated in Rb tissues. Furthermore, the proliferation and migratory and invasive abilities of Rb Y79 and HXO‑RB44 cells were decreased following MIAT downregulation or miR‑665 overexpression. In addition, LASP1 was identified as a target gene of miR‑665. Both the decreased expression of miR‑665 and the elevated expression of LASP1 reversed the suppressive effects of MIAT knockdown on the proliferation and migratory and invasive abilities of Y79 cells. Furthermore, MIAT silencing attenuated the development of Rb by regulating the miR‑665/LASP1 axis. Taken together, these findings suggested that MIAT may be considered as a possible therapeutic target for Rb.

Introduction

Retinoblastoma (Rb) is a common malignant tumour reported mainly in children and affecting ~8,000 infants worldwide annually (1). Rb morbidity rate in developed countries is lower than that observed in underdeveloped countries, suggesting that Rb morbidity is closely associated with effective diagnosis and treatment (2,3). At present, the main treatments of Rb include enucleation, laser photocoagulation, chemotherapy and focal therapy (4). However, the therapeutic effects remain limited due to accelerated metastasis formation in Rb (5). It is therefore crucial to determine effective therapeutic targets for Rb to diminish the formation of metastasis.

Long non-coding RNAs (lncRNAs) are defined as transcripts of >200 nucleotides in length (6). Certain lncRNAs have been reported to contribute considerably to the pathogenesis of Rb, including LINC00152(7), small nucleolar RNA host gene 16 (SNHG16) (8), Pvt1 oncogene (PVT1) (9) and TP73 antisense RNA (TP73-AS1) (10). Yang et al (8) demonstrated that SNHG16 downregulation can restrain Rb cell migratory and invasive abilities. Wu et al (9) reported that PVT1 silencing not only suppresses the proliferation and migratory and invasive abilities of Rb cells in vitro, but also inhibits the growth of tumour xenografts in vivo. Wang et al (10) reported increased expression of TP73-AS1 in Rb tissues and cell lines (Y79, HXO-RB44, WERI-Rb-1 and SO-RB50), whereas TP37-AS1 overexpression could further aggravate the malignant phenotype of Rb in vitro (10). All the aforementioned lncRNAs serve as oncogenes in Rb. In addition, the lncRNA myocardial infarction-associated transcript (MIAT) was demonstrated to facilitate the progression of numerous types of human cancer, including cholangiocarcinoma (CCA) (11), non-small cell lung cancer (NSCLC) (12,13), cervical cancer (CC) (14), ovarian cancer (OC) (15), colorectal cancer (CRC) (16) and gastric cancer (GC) (17). However, the potential role and underlying mechanism of MIAT in Rb remain unclear.

MicroRNAs (miRNAs/miRs) are a class of small endogenous RNA that can target the 3'-untranslated region to regulate gene expression (18). The anti-tumour roles of certain miRNAs in human cancer, especially in Rb, including miR-22-3p (19), miR-124(20), miR-182-5p (8), miR-128-3p (8), miR-488-3p (9), miR-506-3p (21) and miR-936(22), have received increased attention. Furthermore, miR-665 has been reported to serve a crucial role in inhibiting Rb function (23,24). A recent study demonstrated that miR-665 exerts its inhibitory effect on Rb by inactivating the Wnt/β-catenin pathway (23). In addition, miR-665 regulation by LINC00205 has been demonstrated to restrain Rb tumorigenesis (24). The modulation of miR-665 activity by MIAT during Rb progression requires therefore further investigation.

LIM and SH3 protein 1 (LASP1) is an oncogene in the pathogenesis of several human cancers, such as hepatocellular carcinoma (HCC) (25), CRC (26), prostate cancer (27) and breast cancer (28). In these cancers, LASP1 has been indicated to be regulated by miRNAs involved in cancer progression (25,29). Hu et al (25) revealed that miR-326 repressed the cell proliferation and invasion of HCC via directly targeting LASP1. Song et al (29) demonstrated that LASP1 is a downstream target of miR-342-3p affecting the progression of oral squamous cell carcinoma. In addition, Yang et al (8) demonstrated that lncRNA SNHG16 promoted the migration and invasion of Rb cells by regulating LASP1. Furthermore, interestingly, Liu et al (30) indicated that MIAT interacted with LASP1 in the development of papillary thyroid cancer. Nevertheless, the association between miR-665 and LASP1, as well as the role of the MIAT/miR-665/LASP1 axis in the pathogenesis of Rb, are largely unknown.

In the present study, the expression levels of MIAT, miR-665 and LASP1 in Rb tissues and cells were analysed. The detailed regulatory mechanisms of the MIAT/miR-665/LASP1 axis in the pathogenesis of Rb were also explored. The findings from this study may provide a possible therapeutic target for Rb.

Materials and methods

Patients with Rb

A total of 47 patients with Rb (age range, 4 months to 14 years; mean age, 5.54±3.09 years) were enrolled in the No. 215 Hospital of Shaanxi Nuclear Industry (Xianyang, China) between March 2017 and July 2019. The Rb tissues and adjacent normal tissues (1 cm from the edge of the tumour tissues) were collected following an enucleation procedure. Tumour tissues were histologically confirmed and the adjacent tissues without histopathological changes were considered as the normal control group. None of the patients received any preoperative radiotherapy and/or chemotherapy. Each patient or their parents provided written informed consent. The protocol was approved by the Ethics Committee of the No. 215 Hospital of Shaanxi Nuclear Industry (approval no. EC-20200924-1017).

Cell culture and transfection

The human retinal epithelial ARPE-19 cell line and the human Rb Y79, HXO-RB44, WERI-Rb-1 and SO-RB50 cell lines were obtained from the American Type Culture Collection. These cell lines were selected according to previous studies (9,31-34). All cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Invitrogen; Thermo Fisher Scientific, Inc.) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and placed at 37˚C in a humidified incubator containing 5% CO2.

The plasmids used in the present study were synthesised by Hanbio Biotechnology Co., Ltd. and included the short hairpin (sh)RNA-MIAT (sh-MIAT; 5'-UCCUCCGAACCUGGCA CGU-3'), shRNA-negative control (sh-NC; 5'-UUCUCCGAAC GUGUCACGU-3'), miR-665 mimics (5'-ACCAGGAGGCU GAGGCCCCU-3'), mimics-NC (miR-NC; 5'-UUCUCCGAA CGUGUCACGUTT-3'), miR-665 inhibitor (5'-AGGGGCCU CAGCCUCCUGGU-3'), inhibitor NC (5'-CAGUACUUUUGU GUAGUACAA-3'), LASP1 overexpression vector (pcDNA-LASP1) and pcDNA-negative control (pcDNA-NC). Transfections (all the above molecules at 20 nM) were performed using Lipofectamine® 3000 (Thermo Fisher Scientific, Inc.) according to the manufacturers' instructions. After 48 h, cells were collected for subsequent experiments.

Reverse transcription-quantitative (RT-q)PCR

Total RNA was extracted from Rb tissues and the Y79, HXO-RB44, WERI-Rb-1 and SO-RB50 cell lines using TRIzol® (Invitrogen; Thermo Fisher Scientific, Inc.). According to the manufacturer's instructions, RNA was reversed transcribed into cDNA using the First-Strand cDNA Synthesis kit (APeXBIO Technology LLC). RT-qPCR was performed using the SYBR-Green FAST Mastermix (Qiagen GmbH). The following thermocycling conditions were used for the qPCR: Initial denaturation at 95˚C for 3 min, followed by 40 cycles at 95˚C for 15 sec (denaturation), 60˚C for 30 sec (annealing), 72˚C for 1 min (elongation) and a final extension at 72˚C for 5 min. GADPH and U6 were used as internal references. The relative expression levels were normalized to endogenous control and were expressed as 2-ΔΔCq (35). The respective sequences of primers were as follows: MIAT forward, 5'-TCTTCATGTCAGAACACGCTTTA-3' and reverse, 5'-AAGGTCACCCGAGGTCCAA-3'; miR-665 forward, 5'-GCCGAGACCAGGAGGCTGAG-3' and reverse, 5'-CTCAACTGGTGTCGTGGA-3'; LASP1 forward, 5'-GGT GCGGCAAGATCGTGTA-3' and reverse, 5'-TGCAGGTCT CGCAATGGAA-3'; GAPDH forward, 5'-CCAGGTGGTCTC CTCTGA-3' and reverse, 5'-GCTGTAGCCAAATCGTTGT-3'; and U6 forward, 5'-CTCGCTTCGGCAGCACA-3' and reverse, 5'-AACGCTTCACGAATTTGCGT-3'.

MTT assay

Cells were cultured in 96-well plates at a density of 5x104 cells/ml for 96 h, followed by addition of 15 µl MTT (Procell Life Science & Technology, Co., Ltd.) and incubation for 2 h at 37˚C. Subsequently, 100 µl DMSO was added to dissolve the formazan. The optical density was measured at 570 nm using a microplate reader (Thermo Fisher Scientific, Inc.).

Migration and invasion assays

For the migration assay, transfected Y79 and HXO-RB44 cells (5x104 cells/ml) were re-suspended in serum-free DMEM and seeded in the upper chamber of a Transwell insert (8 µm pore size; BD Biosciences). Simultaneously, DMEM containing 10% FBS was added in the lower chamber. For the invasion assay, Matrigel matrix (Becton-Dickinson and Company) was used to coat the membranes before cell seeding. Following overnight incubation at 37˚C, cells in the lower chamber were fixed with 4% paraformaldehyde at 37˚C for 1 h and stained with 0.1% crystal violet for 15 min at 37˚C. The stained cells were imaged using an inverted light microscope (Olympus Corporation) and analysed with ImageJ software (version 1.46; National Institutes of Health).

Dual luciferase reporter (DLR) assay

The targeting association between MIAT and miR-665 was analyzed using the StarBase software (version 2.0; http://starbase.sysu.edu.cn). Additionally, the targeting interaction between miR-665 and LASP1 was predicted using TargetScan software (v7.2; http://www.targetscan.org/vert_72/). The predicted binding region sequences (MIAT, 5'-GGGUUCCAGGCUCCUGG-3'; LASP1, 5'-CUCCUGGU-3') were inserted into pGL3 vector (Promega Corporation) to construct the wild-type (wt) phenotype. For construction of the mutant (mut) phenotype, the mutation sequences (MIAT mut, 5'-CCCUUCGUCGGAGGACC-3'; LASP1 mut, 5'-GAGGACCA-3') were inserted into pGL3. Y79 and HXO-RB44 cells were then co-transfected with MIAT/LASP1-wt or MIAT/LASP1-mut (80 ng) and miR-665 mimics/miR-NC (50 nM) using Lipofectamine 3000 (Invitrogen; Thermo Fisher Scientific, Inc.) at 37˚C for 48 h, followed by detection of the luciferase activity using a dual-luciferase reporter assay system (Promega Corporation). The activity of firefly luciferase was normalized to that of Renilla luciferase.

Western blotting

Transfected cells were lysed on ice with RIPA (Beyotime Institute of Biotechnology) containing 10 mmol/l PMSF (Beyotime Institute of Biotechnology). The protein concentration was detected using a BCA Protein Assay Kit (Abcam). A total of 50 µg of protein/lane was separated via 10% SDS-PAGE and transferred onto PVDF membranes. Following blocking with 5% skimmed milk for 2 h at 25˚C, membranes were incubated with the primary antibodies against LASP1 (1:1,000; cat. no. SAB1402251; Sigma-Aldrich; Merck KGaA) and α-tubulin (1:1,000; cat. no. T6199; Sigma-Aldrich; Merck KGaA) at 4˚C overnight. Membranes were then incubated with the HRP-conjugated anti-mouse IgG secondary antibody (1:5,000; cat. no. sc-2005; Santa Cruz Biotechnology, Inc.) for 1 h at 37˚C. Bands were detected using enhanced chemiluminescence substrate (Amersham; Cytiva). Relative expression level of LASP1 was normalized to endogenous control α-tubulin. The immunoblots were visualized using an ECL detection kit (Thermo Fisher Scientific, Inc.) using Gel-Pro analyzer (version 4.0; Media Cybernetics, Inc.).

Statistical analysis

The SPSS 20.0 software (IBM Corp.) was used to perform statistical analysis. All cell experiments were performed three times. Data are presented as the mean ± standard deviation. Student's t-test was used to assess the differences between two groups. One-way ANOVA followed by Tukey's post hoc test was used to evaluate the differences among multiple groups. χ2 test was used to analyse data from Table I. The linear correlation was assessed by Pearson's correlation analysis. P<0.05 was considered to indicate a statistically significant difference.

Table I

Association between MIAT expression and the clinicopathological characteristics of patients with retinoblastoma.

Table I

Association between MIAT expression and the clinicopathological characteristics of patients with retinoblastoma.

 MIAT expression, n 
CharacteristicsTotal, nLow (n=23)High (n=24)P-value
Age, years   0.192
     <520128 
     ≥5271116 
Sex   0.891
     Male221111 
     Female251213 
Tumor size, mm   0.440
     <10281513 
     ≥1019811 
Optic nerve invasion   0.044
     No301812 
     Yes17512 
TNM stage   0.012
     I/II18135 
     III/IV291019 
IIRC stage   0.0035
     Early stages (A-C)15123 
     Advanced stages (D and E)321121 

[i] TNM, Tumor-Node-Metastasis; IIRC, intraocular international retinoblastoma classification; Rb, retinoblastoma; MIAT, MIAT, myocardial infarction-associated transcript.

Results

High MIAT expression is observed in Rb tissues and cells

The expression of MIAT was initially detected in Rb tissues using RT-qPCR. The results demonstrated that MIAT expression was upregulated in Rb tissues compared with that in adjacent tissues (Fig. 1A; P<0.01). Furthermore, a significantly increased expression level of MIAT was observed in the tissues from patients with Tumour-Node-Metastasis (TNM) stage III/IV compared with that in the tissues from patients with TNM stage I/II (Fig. 1B; P<0.01). As shown in Table I, MIAT expression was significantly associated with intraocular international retinoblastoma classification (IIRC) stage (P=0.0035), TNM stage (P=0.012) and optic nerve invasion (P=0.044). In addition, MIAT mRNA expression was detected in the Rb WERI-Rb-1, SO-RB50, HXO-RB44 and Y79 cell lines, and the normal ARPE-19 cell line. The results from RT-qPCR demonstrated that MIAT expression level was significantly increased in all Rb cell lines compared with that in the ARPE-19 cell line (Fig. 1C; P<0.01). Since the Y79 and HXO-RB44 cell lines exhibited the highest MIAT expression levels amongst all Rb cell lines, they were selected for subsequent experiments.

MIAT knockdown inhibits the proliferation and migratory and invasive abilities of Rb cells in vitro

To explore the effects of MIAT on Rb progression in vitro, Y79 and HXO-RB44 cells were transfected with sh-MIAT or sh-NC and the transfection efficiency was detected by RT-qPCR. The results demonstrated that the expression level of MIAT was significantly decreased following transfection with sh-MIAT compared with sh-NC, which confirmed the successful transfection into Y79 and HXO-RB44 cells (Fig. 2A; P<0.01). Subsequently, transfected Y79 and HXO-RB44 cells were used to determine the proliferation and migratory and invasive abilities of Rb cells using MTT assay and Transwell assay. As presented in Fig. 2B, Y79 and HXO-RB44 cell proliferation was significantly decreased following transfection with sh-MIAT compared with that in the sh-NC group (P<0.01). Furthermore, both migratory and invasive abilities of Rb cell lines were decreased in the sh-MIAT group compared with those in the sh-NC group (Fig. 2C and D; P<0.01).

MIAT acts as an endogenous sponge of miR-665

lncRNAs can act as competing endogenous RNAs or sponges of miRNAs to modulate cancer progression (36). Through the StarBase software, 58 target miRNAs of MIAT were predicted (data not shown). In the present study, miR-665 was selected due to its important role in Rb (23,24) (Fig. 3A). A DLR assay was used to confirm the target relationship between MIAT and miR-665 in Y79 and HXO-RB44 cells. The results demonstrated that the luciferase activity in MIAT wt/miR-665 mimics was decreased, which was not the case in the MIAT mut (Fig. 3B; P<0.01). The results from RT-qPCR indicated that the expression levels of MIAT and miR-665 in Rb cell lines (Fig. 3C; P<0.01) and Rb tissues (Fig. 3E; P=0.001, r=-0.4614) were negatively correlated. In addition, a decreased expression level of miR-665 was detected in Rb tissues compared with that in the adjacent tissues (Fig. 3D; P<0.01).

High expression of miR-665 serves as a suppressor of the proliferation and migratory and invasive abilities of Rb cells in vitro

Both miR-665 mimics and miR-665 inhibitor were transfected into Y79 and HXO-RB44 cells, and the effects of miR-665 overexpression on Rb progression in vitro were subsequently evaluated. The results demonstrated that miR-665 was significantly upregulated following transfection with miR-665 mimics, whereas it was significantly downregulated following transfection with miR-665 inhibitor (Fig. 4A; P<0.01). Similar to the effects of MIAT knockdown on the progression of Rb in vitro, the results from the MTT assay and Transwell assay revealed that the proliferation and migratory and invasive abilities of Rb cells were inhibited following miR-665 overexpression (Fig. 4B-D; P<0.01).

LASP1 is a downstream target gene of miR-665

As illustrated in Fig. 5A, the TargetScan software predicted a binding site between miR-665 and LASP1. A DLR assay was subsequently used to confirm the binding relationship, and a decrease in luciferase activity in the LASP1 wt/miR-665 mimics groups was observed in both Y79 and HXO-RB44 cells (Fig. 5B; P<0.01). Furthermore, LASP1 was overexpressed in Rb tissues compared with that in adjacent tissues (Fig. 5C; P<0.01). In addition, the results from Pearson's correlation analysis demonstrated a negative correlation between LASP1 and miR-665 expression (Fig. 5D; P<0.01, r=-0.4738); however, a positive correlation was observed between LASP1 and MIAT expression (Fig. 5E; P<0.01, r=0.4116) in Rb tissues. To further verify the interaction between the expression of miR-665 and LASP1, western blotting was performed to determine the protein expression of LASP1 following transfection of Y79 and HXO-RB44 cells with miR-665 mimics. The results revealed that LASP1 protein expression was decreased following miR-665 upregulation (Fig. 5F; P<0.01).

MIAT knockdown decreases the development of Rb by sponging miR-665 and regulating LASP1

The results from western blotting revealed that LASP1 expression was increased following transfection with pcDNA-LASP1 (Fig. 6A; P<0.01). The results from MTT assay and Transwell assay demonstrated that the suppressive effects of sh-MIAT on the proliferation and migratory and invasive abilities of Y79 and HXO-RB44 cells (Fig. 2) were reversed following transfection with miR-665 inhibitor or pcDNA-LASP1 (Fig. 6B-D; P<0.01).

Discussion

Chemotherapy and radiation are not the optimal therapy strategies for Rb as most of the patients with Rb are children or infants and these therapies can cause serious physical injury (8). Determining a novel therapeutic target for Rb is therefore essential. Growing evidence has demonstrated that certain lncRNAs are upregulated in Rb and are associated with some pathological features of Rb, such as the association between XIST and TNM stage (34), the association of AFAP1-AS1 with choroidal invasion and optic nerve invasion (37), the association of SNHG16 with choroidal invasion, optic nerve invasion and TNM stage (8), and the association of PVT1 with optic nerve invasion and IIRC stage (9). The present study demonstrated that MIAT was overexpressed in Rb tissues and cell lines. Furthermore, MIAT expression was significantly associated with IIRC stage, TNM stage and optic nerve invasion. These findings suggested that MIAT may be considered as a risk factor in Rb and therefore a potential target for treating Rb.

To further explore the possible effect of MIAT on the pathogenesis of Rb, sh-MIAT was transfected into Y79 and HXO-RB44 cells. The results demonstrated that the proliferation and migratory and invasive abilities of Rb cells were suppressed following MIAT knockdown. Similarly, MIAT has been reported to slow tumorigenesis in several types of human cancer, such as CCA, NSCLC and CC (11-14). Chang et al (11) demonstrated that MIAT silencing can inhibit the proliferation and accelerate the apoptosis of CCA cells. Similar to the findings reported by Zhou et al (12), Li et al (13) reported that transfection with sh-MIAT inhibits the proliferation, migratory and invasive abilities of NSCLC cells. Similar results were reported in a study by Zhang et al (14), which revealed that MIAT downregulation has an inhibitory effect on the proliferation and migration of CC cells in vitro (14). The present study therefore hypothesized that MIAT silencing may inhibit the progression of Rb.

Increasing research efforts have been engaged to elucidate the anti-tumour roles of miR-665 in human cancer (38-40). For example, miR-665 downregulation was reported to reverse the suppressive effects of the lncRNA RHPN1-AS1 on the proliferation and migratory and invasive abilities of OC cells (38). Furthermore, transfection with miR-665 inhibitor partly aggravates the tumorigenicity of CRC in vitro (39). A recent study by Wu et al (40) revealed that miR-665 expression is downregulated in GC tissues, and that miR-665 overexpression exhibits a visible effect on tumour inhibition. In the present study, a decreased expression of miR-665 was found in Rb tissues, and miR-665 overexpression significantly reduced the proliferation and migratory and invasive abilities of Rb cells. Similarly, Wang et al (23) reported that miR-665 is minimally expressed in Rb tissues and cell lines, and that the proliferation and migratory and invasive abilities of Rb cells are inhibited following transfection with miR-665 mimics. In the present study, MIAT was shown to target and negatively modulate miR-665 expression, suggesting that miR-665 may be involved in Rb tumorigenesis via MIAT regulation. The results of the present study further demonstrated that the decreased expression of miR-665 reversed the inhibitory effects of MIAT silencing on the proliferation and migratory and invasive abilities of Y79 cells. These findings indicated that MIAT knockdown may attenuate Rb malignancy by modulating miR-665.

LASP1, an oncogene involved in cancer aggressiveness (41), was found to be upregulated in numerous types of cancer, including gallbladder cancer (42), NSCLC (43), CRC (44) and OC (45). In the present study, LASP1 was demonstrated to be highly expressed in Rb tissues compared with that in adjacent tissues. Consistent with these findings, Yang et al (8) reported a high expression level of LASP1 in Rb tissues. These results suggested that LASP1 may act as an oncogene in Rb pathogenesis. Furthermore, a positive correlation between MIAT and LASP1 expression was observed in the present study. Similarly, Liu et al (46) confirmed that the expression of LASP1 is positively correlated with MIAT expression in papillary thyroid cancer tissues. Furthermore, the present study demonstrated that LASP1 may be a target gene of miR-665 and was negatively regulated by miR-665. These findings suggested that miR-665 may be involved in the tumorigenesis of Rb via MIAT regulation. We hypothesized that LASP1 may be modulated by MIAT and be involved in Rb progression. The feedback verification experiments demonstrated that the enhancement of proliferation and migratory and invasive abilities of Y79 cells caused by MIAT silencing were restrained following transfection with pcDNA-LASP1, which confirmed this hypothesis. These findings indicated that MIAT downregulation may reduce the progression of Rb by sponging miR-665 and regulating LASP1.

In summary, the present study reported an elevated expression level of MIAT in Rb tissues and demonstrated that MIAT silencing significantly reduced the tumorigenicity of Rb by modulating the miR-665/LASP1 axis in vitro. The results from this study provide a novel target for treating Rb and may be applied in the clinical setting. One limitation of this study is the absence of in vivo experiments to confirm the interactions among MIAT, miR-665 and LASP1. Further investigation will therefore be performed in the future.

Acknowledgements

Not applicable.

Availability of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Authors' contributions

XX and BD made significant contributions to the overall structure design of the study, data analysis, methodology, project management, funding and draft writing. YZ and GD are mainly responsible for resource collection and integration, experiments, experimental data analysis, software processing and paper modification and editing. All authors confirmed the authenticity of all the raw data and have read and approved the final manuscript.

Ethics approval and consent to participate

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of No. 215 Hospital of Shaanxi Nuclear Industry (approval no. EC-20200924-1017). Each patient or their parents provided written informed consent.

Patient consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

References

1 

Pascual-Pasto G, Bazan-Peregrino M, Olaciregui NG, Restrepo-Perdomo CA, Mato-Berciano A, Ottaviani D, Weber K, Correa G, Paco S, Vila-Ubach M, et al: Therapeutic targeting of the RB1 pathway in retinoblastoma with the oncolytic adenovirus VCN-01. Sci Transl Med. 11(11)2019.PubMed/NCBI View Article : Google Scholar

2 

Chantada GL, Qaddoumi I, Canturk S, Khetan V, Ma Z, Kimani K, Yeniad B, Sultan I, Sitorus RS, Tacyildiz N, et al: Strategies to manage retinoblastoma in developing countries. Pediatr Blood Cancer. 56:341–348. 2011.PubMed/NCBI View Article : Google Scholar

3 

Canturk S, Qaddoumi I, Khetan V, Ma Z, Furmanchuk A, Antoneli CB, Sultan I, Kebudi R, Sharma T, Rodriguez-Galindo C, et al: Survival of retinoblastoma in less-developed countries impact of socioeconomic and health-related indicators. Br J Ophthalmol. 94:1432–1436. 2010.PubMed/NCBI View Article : Google Scholar

4 

Errico A: Cancer therapy: Retinoblastoma - chemotherapy increases the risk of secondary cancer. Nat Rev Clin Oncol. 11(623)2014.PubMed/NCBI View Article : Google Scholar

5 

Shields CL, Say EA, Pointdujour-Lim R, Cao C, Jabbour PM and Shields JA: Rescue intra-arterial chemotherapy following retinoblastoma recurrence after initial intra-arterial chemotherapy. J Fr Ophtalmol. 38:542–549. 2015.PubMed/NCBI View Article : Google Scholar

6 

Rao AKDM, Rajkumar T and Mani S: Perspectives of long non-coding RNAs in cancer. Mol Biol Rep. 44:203–218. 2017.PubMed/NCBI View Article : Google Scholar

7 

Li S, Wen D, Che S, Cui Z, Sun Y, Ren H and Hao J: Knockdown of long noncoding RNA 00152 (LINC00152) inhibits human retinoblastoma progression. OncoTargets Ther. 11:3215–3223. 2018.PubMed/NCBI View Article : Google Scholar

8 

Yang L, Zhang L, Lu L and Wang Y: Long noncoding RNA SNHG16 sponges miR-182-5p and miR-128-3p to promote retinoblastoma cell migration and invasion by targeting LASP1. OncoTargets Ther. 12:8653–8662. 2019.PubMed/NCBI View Article : Google Scholar

9 

Wu XZ, Cui HP, Lv HJ and Feng L: Knockdown of lncRNA PVT1 inhibits retinoblastoma progression by sponging miR-488-3p. Biomed Pharmacother. 112(108627)2019.PubMed/NCBI View Article : Google Scholar

10 

Wang L, Wang C, Wu T and Sun F: Long non-coding RNA TP73-AS1 promotes TFAP2B-mediated proliferation, metastasis and invasion in retinoblastoma via decoying of miRNA-874-3p. J Cell Commun Signal. 14:193–205. 2020.PubMed/NCBI View Article : Google Scholar

11 

Chang W, Wang Y, Li W and Geng Z: Long non-coding RNA myocardial infarction associated transcript promotes the proliferation of cholangiocarcinoma cells by targeting miR-551b-3p/CCND1 axis. Clin Exp Pharmacol Physiol. 47:1067–1075. 2020.PubMed/NCBI View Article : Google Scholar

12 

Zhou Z, Zhang S and Xiong Y: Long noncoding RNA MIAT promotes non-small cell lung cancer progression by sponging miR-149-5p and regulating FOXM1 expression. Cancer Cell Int. 20(348)2020.PubMed/NCBI View Article : Google Scholar

13 

Li F, Li H, Li S, Lv B, Shi J, Yan H, Zhang H and He Y: Long Non-coding RNA MIAT mediates non-small cell lung cancer development through regulating the miR-128-3p/PELI3 Axis. Biochem Genet. 58:867–882. 2020.PubMed/NCBI View Article : Google Scholar

14 

Zhang L, Ge S and Cao B: Long non-coding RNA MIAT promotes cervical cancer proliferation and migration. J Biochem. 168:183–190. 2020.PubMed/NCBI View Article : Google Scholar

15 

Zhou S, Xu A, Song T, Gao F, Sun H and Kong X: lncRNA MIAT regulates cell growth, migration, and invasion through sponging miR-150-5p in ovarian cancer. Cancer Biother Radiopharm. 35:650–660. 2020.PubMed/NCBI View Article : Google Scholar

16 

Liu Y, Peng H, Shen Y, Da R, Tian A and Guo X: Downregulation of long noncoding RNA myocardial infarction associated transcript suppresses cell proliferation, migration, invasion, and glycolysis by regulation of miR-488-3p/IGF1R pathway in colorectal cancer. Cancer Biother Radiopharm: Oct 21, 2020 (Epub ahead of print). doi: 10.1089/cbr.2020.3671.

17 

Sha M, Lin M, Wang J, Ye J, Xu J, Xu N and Huang J: Long non-coding RNA MIAT promotes gastric cancer growth and metastasis through regulation of miR-141/DDX5 pathway. J Exp Clin Cancer Res. 37(58)2018.PubMed/NCBI View Article : Google Scholar

18 

Lu TX and Rothenberg ME: MicroRNA. J Allergy Clin Immunol. 141:1202–1207. 2018.PubMed/NCBI View Article : Google Scholar

19 

Liu Y, Li H, Liu Y and Zhu Z: MiR-22-3p targeting alpha-enolase 1 regulates the proliferation of retinoblastoma cells. Biomed Pharmacother. 105:805–812. 2018.PubMed/NCBI View Article : Google Scholar

20 

Wang L, Yang D, Tian R and Zhang H: NEAT1 promotes retinoblastoma progression via modulating miR-124. J Cell Biochem. 120:15585–15593. 2019.PubMed/NCBI View Article : Google Scholar

21 

Han N, Zuo L, Chen H, Zhang C, He P and Yan H: Long non-coding RNA homeobox A11 antisense RNA (HOXA11-AS) promotes retinoblastoma progression via sponging miR-506-3p. OncoTargets Ther. 12:3509–3517. 2019.PubMed/NCBI View Article : Google Scholar

22 

Xu L, Li W, Shi Q, Wang M, Li H, Yang X and Zhang J: MicroRNA 936 inhibits the malignant phenotype of retinoblastoma by directly targeting HDAC9 and deactivating the PI3K/AKT pathway. Oncol Rep. 43:635–645. 2020.PubMed/NCBI View Article : Google Scholar

23 

Wang S, Du S, Lv Y, Zhang F and Wang W: MicroRNA-665 inhibits the oncogenicity of retinoblastoma by directly targeting high-mobility group box 1 and inactivating the Wnt/β-catenin pathway. Cancer Manag Res. 11:3111–3123. 2019.PubMed/NCBI View Article : Google Scholar

24 

Zhang S, Long J and Hu Y: Long noncoding RNA LINC00205 enhances the malignant characteristics of retinoblastoma by acting as a molecular sponge of microRNA-665 and consequently increasing HMGB1 expression. Biochem Biophys Res Commun. 526:396–403. 2020.PubMed/NCBI View Article : Google Scholar

25 

Hu S, Ran Y, Chen W, Zhang Y and Xu Y: MicroRNA-326 inhibits cell proliferation and invasion, activating apoptosis in hepatocellular carcinoma by directly targeting LIM and SH3 protein 1. Oncol Rep. 38:1569–1578. 2017.PubMed/NCBI View Article : Google Scholar

26 

Yan P, Liu J, Zhou R, Lin C, Wu K, Yang S, Yang S, Zhou J, Xu L, Wang H, et al: LASP1 interacts with N-WASP to activate the Arp2/3 complex and facilitate colorectal cancer metastasis by increasing tumour budding and worsening the pattern of invasion. Oncogene. 39:5743–5755. 2020.PubMed/NCBI View Article : Google Scholar

27 

Wang S, Qiu J, Wang L, Wu Z, Zhang X, Li Q and Jiang F: Long non-coding RNA LINC01207 promotes prostate cancer progression by downregulating microRNA-1972 and upregulating LIM and SH3 protein 1. IUBMB Life. 72:1960–1975. 2020.PubMed/NCBI View Article : Google Scholar

28 

Sui Y, Zhang X, Yang H, Wei W and Wang M: MicroRNA-133a acts as a tumour suppressor in breast cancer through targeting LASP1. Oncol Rep. 39:473–482. 2018.PubMed/NCBI View Article : Google Scholar

29 

Song X, Jin Y, Yan M, Zhang Y and Chen B: MicroRNA-342-3p functions as a tumor suppressor by targeting LIM and SH3 protein 1 in oral squamous cell carcinoma. Oncol Lett. 17:688–696. 2019.PubMed/NCBI View Article : Google Scholar

30 

Liu W, Wang ZL, Wang C and Ai ZL: Long non-coding RNA MIAT promotes papillary thyroid cancer progression through upregulating LASP1. Cancer Cell Int. 19(194)2019.PubMed/NCBI View Article : Google Scholar

31 

Wang JX, Yang Y and Li K: Long noncoding RNA DANCR aggravates retinoblastoma through miR-34c and miR-613 by targeting MMP-9. J Cell Physiol. 233:6986–6995. 2018.PubMed/NCBI View Article : Google Scholar

32 

Shang W, Yang Y, Zhang J and Wu Q: Long noncoding RNA BDNF-AS is a potential biomarker and regulates cancer development in human retinoblastoma. Biochem Biophys Res Commun. 497:1142–1148. 2018.PubMed/NCBI View Article : Google Scholar

33 

Yan G, Su Y, Ma Z, Yu L and Chen N: Long Noncoding RNA LINC00202 promotes tumor progression by sponging miR-3619-5p in retinoblastoma. Cell Struct Funct. 44:51–60. 2019.PubMed/NCBI View Article : Google Scholar

34 

Hu C, Liu S, Han M, Wang Y and Xu C: Knockdown of lncRNA XIST inhibits retinoblastoma progression by modulating the miR-124/STAT3 axis. Biomed Pharmacother. 107:547–554. 2018.PubMed/NCBI View Article : Google Scholar

35 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar

36 

Du Z, Sun T, Hacisuleyman E, Fei T, Wang X, Brown M, Rinn JL, Lee MG, Chen Y, Kantoff PW, et al: Integrative analyses reveal a long noncoding RNA-mediated sponge regulatory network in prostate cancer. Nat Commun. 7(10982)2016.PubMed/NCBI View Article : Google Scholar

37 

Hao F, Mou Y, Zhang L, Wang S and Yang Y: LncRNA AFAP1-AS1 is a prognostic biomarker and serves as oncogenic role in retinoblastoma. Biosci Rep. 38(38)2018.PubMed/NCBI View Article : Google Scholar

38 

Zhao J, Yang T, Ji J, Zhao F, Li C and Han X: RHPN1-AS1 promotes cell proliferation and migration via miR-665/Akt3 in ovarian cancer. Cancer Gene Ther. 28:33–41. 2021.PubMed/NCBI View Article : Google Scholar

39 

Ouyang S, Zhou X, Chen Z, Wang M, Zheng X and Xie M: LncRNA BCAR4, targeting to miR-665/STAT3 signaling, maintains cancer stem cells stemness and promotes tumorigenicity in colorectal cancer. Cancer Cell Int. 19(72)2019.PubMed/NCBI View Article : Google Scholar

40 

Wu KZ, Zhang CD, Zhang C, Pei JP and Dai DQ: miR-665 suppresses the epithelial-mesenchymal transition and progression of gastric cancer by targeting CRIM1. Cancer Manag Res. 12:3489–3501. 2020.PubMed/NCBI View Article : Google Scholar

41 

Orth MF, Cazes A, Butt E and Grunewald TG: An update on the LIM and SH3 domain protein 1 (LASP1): A versatile structural, signaling, and biomarker protein. Oncotarget. 6:26–42. 2015.PubMed/NCBI View Article : Google Scholar

42 

Li Z, Chen Y, Wang X, Zhang H, Zhang Y, Gao Y, Weng M, Wang L, Liang H, Li M, et al: LASP-1 induces proliferation, metastasis and cell cycle arrest at the G2/M phase in gallbladder cancer by down-regulating S100P via the PI3K/AKT pathway. Cancer Lett. 372:239–250. 2016.PubMed/NCBI View Article : Google Scholar

43 

Zheng J, Wang F, Lu S and Wang X: LASP-1, regulated by miR-203, promotes tumor proliferation and aggressiveness in human non-small cell lung cancer. Exp Mol Pathol. 100:116–124. 2016.PubMed/NCBI View Article : Google Scholar

44 

Zhao L, Wang H, Liu C, Liu Y, Wang X, Wang S, Sun X, Li J, Deng Y, Jiang Y, et al: Promotion of colorectal cancer growth and metastasis by the LIM and SH3 domain protein 1. Gut. 59:1226–1235. 2010.PubMed/NCBI View Article : Google Scholar

45 

Grunewald TG, Kammerer U, Winkler C, Schindler D, Sickmann A, Honig A and Butt E: Overexpression of LASP-1 mediates migration and proliferation of human ovarian cancer cells and influences zyxin localisation. Br J Cancer. 96:296–305. 2007.PubMed/NCBI View Article : Google Scholar

46 

Liu W, Wang Z, Wang C and Ai Z: Long non-coding RNA MIAT promotes papillary thyroid cancer progression through upregulating LASP1. Cancer Cell Int. 19(194)2019.PubMed/NCBI View Article : Google Scholar

Related Articles

Journal Cover

November-2021
Volume 22 Issue 5

Print ISSN: 1792-0981
Online ISSN:1792-1015

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Xu X, Zhao Y, Duan G and Du B: Downregulation of MIAT reduces the proliferation and migratory and invasive abilities of retinoblastoma cells by sponging miR‑665 and regulating LASP1. Exp Ther Med 22: 1342, 2021
APA
Xu, X., Zhao, Y., Duan, G., & Du, B. (2021). Downregulation of MIAT reduces the proliferation and migratory and invasive abilities of retinoblastoma cells by sponging miR‑665 and regulating LASP1. Experimental and Therapeutic Medicine, 22, 1342. https://doi.org/10.3892/etm.2021.10777
MLA
Xu, X., Zhao, Y., Duan, G., Du, B."Downregulation of MIAT reduces the proliferation and migratory and invasive abilities of retinoblastoma cells by sponging miR‑665 and regulating LASP1". Experimental and Therapeutic Medicine 22.5 (2021): 1342.
Chicago
Xu, X., Zhao, Y., Duan, G., Du, B."Downregulation of MIAT reduces the proliferation and migratory and invasive abilities of retinoblastoma cells by sponging miR‑665 and regulating LASP1". Experimental and Therapeutic Medicine 22, no. 5 (2021): 1342. https://doi.org/10.3892/etm.2021.10777