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.2022 Oct 21;24(6):438.
doi: 10.3892/ol.2022.13558. eCollection 2022 Dec.

LINC01635, a long non-coding RNA with a cancer/testis expression pattern, promotes lung cancer progression by sponging miR-455-5p

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LINC01635, a long non-coding RNA with a cancer/testis expression pattern, promotes lung cancer progression by sponging miR-455-5p

Wenyi Shen et al. Oncol Lett..

Abstract

Long non-coding RNAs (lncRNAs) have been reported to play vital roles in human lung cancer. In recent years, cancer/testis (CT) lncRNAs have been characterized as a novel class of lncRNA. However, this class of lncRNA remains to be thoroughly investigated. The present study identified long intergenic non-protein coding RNA 1635 (LINC01635), which was highly expressed in testis and in a broad range of human cancer types. Next, it was confirmed that LINC01635 was upregulated significantly in samples from patients with lung cancer and in non-small cell lung carcinoma (NSCLC) cell lines. Silencing LINC01635 suppressed the proliferation and metastasis of NSCLC cellsin vitro andin vivo. Furthermore, it was found that LINC01635 could bind to microRNA (miRNA or miR)-455-5p and regulate the expression of a series of miR-455-5p-targeting tumor-related genes. Knockdown of miR-455-5p partially rescued the progression of lung cancer cells that was suppressed by LINC01635 silencing. Together, the current results demonstrated that LINC01635 may play important roles in NSCLC progression by targeting miR-455-5p, and that it could be a biomarker and therapeutic target for lung cancer.

Keywords: cancer/testis long non-coding RNAs; long intergenic non-protein coding RNA 1635; long non-coding RNA; microRNA-455-5p; non-small cell lung carcinoma.

Copyright: © Shen et al.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1.
Figure 1.
LINC01635 is a long non-coding RNA with a cancer/testis expression pattern. (A) The volcano map of differentially expression genes of the GSE113852 dataset, which contains the expression profiles of 27 lung adenocarcinoma and 27 normal lung tissue samples. The green dots represent downregulated genes (P<0.05), the blue dots represent upregulated genes (P<0.05) and the red dots represent upregulated genes (P<0.05) for which the value of log2(FC) was >2. The log2(FC) value of LINC01635 was 2.36, and the P-value was 8.58×10-8. (B) Non-coding property prediction of LINC01635, including CPAT, CNCI and Secondary Structure Prediction tools. (C) Expression in RPKM of LINC01635 among the Human Protein Atlas RNA-seq normal tissue datasets. (D) The overexpression of LINC01635 in lung cancer tissues was confirmed by analyzing GSE101929 datasets using lnCAR online tools. (E) Expression of LINC01635 in tumor and normal tissue samples from GTEx and TCGA database. *P<0.05; ***P<0.001. LINC01635, long intergenic non-protein coding RNA 1635; CPAT, coding-potential assessment tool; CNCI, coding-non-coding index; RPKM, reads per kilobase million; RNA-seq, RNA-sequencing; lnCAR, lncRNAs from cancer arrays; TCGA, The Cancer Genome Atlas; KICH, kidney chromophobe; ACC, adrenocortical carcinoma; HNSC, head and neck squamous cell carcinoma; PCPG, pheochromocytoma and paraganglioma; BRCA, breast invasive carcinoma; CESC, cervical squamous cell carcinoma and endocervical adenocarcinoma; LIHC, liver hepatocellular carcinoma; READ, rectal adenocarcinoma; BLCA, bladder urothelial carcinoma; CHOL, cholangiocarcinoma; COAD, colon adenocarcinoma; KIRC, kidney renal clear cell carcinoma; PRAD, prostate adenocarcinoma; ESCA, esophageal carcinoma; LGG, brain lower grade glioma; UCEC, uterine corpus endometrial carcinoma; LUSC, lung squamous cell carcinoma; KIRP, kidney renal papillary cell carcinoma; OV, ovarian serous cystadenocarcinoma; UCS, uterine carcinosarcoma; LUAD, lung adenocarcinoma; PAAD, pancreatic adenocarcinoma; THCA, thyroid carcinoma; GBM, glioblastoma multiforme; SARC, sarcoma; TGCT, testicular germ cell tumor; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; THYM, thymoma; DLBC, lymphoid neoplasm diffuse large B-cell lymphoma; LAML, acute myeloid leukemia.
Figure 2.
Figure 2.
LINC01635 expression is upregulated in NSCLC tissue samples and cell lines. (A) The expression of LINC01635 was significantly increased in NSCLC tissue samples compared with that in adjacent normal lung tissue samples according to RT-qPCR analysis (***P<0.001). (B) The expression of LINC01635 was overexpressed in NSCLC cell lines (A549, H1299 and H1975) compared with that in the normal human bronchial epithelial cell line (HBE135) (**P<0.01 and ***P<0.001 vs. HBE135). (C) Transcripts of LINC01635 were examined by RT-PCR in the A549 cell line, which showed two specific bands. (D) Transcripts of LINC01635 were analyzed according to sequencing reports of all RT-PCR products. Blue boxes represent exons, and polylines represent splicing sites. NSCLC, non-small cell lung cancer; LINC01635, long intergenic non-protein coding RNA 1635; RT-qPCR, reverse transcription-quantitative PCR.
Figure 3.
Figure 3.
Knockdown of LINC01635 decreases the proliferation and migration in lung cancer cells. (A and B) The knockdown efficiencies of LINC01635 in (A) A549 and (B) H1975 cell lines were examined by reverse transcription-qPCR. (C and D) CCK-8 assays showed that knockdown of LINC01635 decreased the proliferation of (C) A549 and (D) H1975 cells. (E and F) Transwell assays showed knockdown of LINC01635 decreased the migration of (E) A549 and (F) H1975 cells. (G and H) The expression of (G) MMP2 and (H) MMP9 decreased when LINC01635 was knocked down, as assessed by qPCR. (I) The expression of (G) MMP2 and (H) MMP9 decreased when LINC01635 was knocked down, as assessed by western blotting. Scale bar, 100 µm. *P<0.05, **P<0.01, ***P<0.001. qPCR, quantitative PCR; si, small interfering; NC, negative control; LINC01635, long intergenic non-protein coding RNA 1635.
Figure 4.
Figure 4.
Knockdown of LINC01635 decreases the proliferation and metastasis of lung cancer cells in a zebrafish xenograft. (A-F) Zebrafish xenograft models showed that knockdown of LINC01635 decreased the proliferation and metastasis of A549 cells at 4 dpi. A549 cells transfected with (A and D) NC and (B and E) si-LINC01635 were labeled with CM-DiI and then injected into the PVS of 2-days post-fertilization Tg(fli1a:EGFP) transgenic zebrafish larvae, which labeled the zebrafish endothelial cells. Confocal images were taken at 4 dpi. (A and B) The tumor cells in the PVS were quantified for cell proliferation, (D and E) and the tumor cells in the trunk were quantified for cell metastasis. Statistical analysis of (C) proliferation and (F) metastasis when knocking down LINC01635 in A549 cells compared with the NC. (G-L) Zebrafish xenograft models showed that knockdown of LINC01635 decreased the proliferation and metastasis of H1975 cells. Scale bar, 100 µm. **P<0.01. dpi, days post-injection; si, small interfering; NC, negative control; LINC01635, long intergenic non-protein coding RNA 1635; PVS, perivitelline space.
Figure 5.
Figure 5.
LINC01635 can bind with miR-455-5p and regulate the expression of miR-455-5p-targeting tumor-related genes. (A) LINC01635 subcellular location in A549 was assessed by RT-qPCR after nuclear and cytosolic separation of total RNA. GAPDH was used as a cytosol marker and U6 was used as a nucleus marker. (B) miRNA binding sites of LINC01635 were predicted by miRDB and LncBase tools; 9 miRNAs were predicted by both tools and miR-455-5p had the highest potential for binding with LINC01635. (C) Binding site between LINC01635 transcripts and miR-455-5p, and the LINC01635 with the binding site mutant (LINC01635-mut). The red arrow represents the site of the miR-455-5p targeting site. (D) Dual-luciferase reporter assay showed that LINC01635 could bind with miR-455-5p directly. The relative luciferase reporter activity was reduced when co-transfecting with miR-455-5p mimic and LINC01635 reporter plasmid, but not LINC01635-mut reporter plasmid (*P<0.05). (E) The expression level of miR-455-5p-targeting tumor-related genes was examined by RT-qPCR following knockdown of LINC01635 or overexpression of miR-455-5p (*P<0.05 and ***P<0.001 vs. NC). miR/miRNA, microRNA; si, small interfering; NC, negative control; LINC01635, long intergenic non-protein coding RNA 1635; RT-qPCR, reverse transcription-quantitative PCR; WT, wild-type; mut, mutant.
Figure 6.
Figure 6.
Knockdown of miR-455-5p partially rescues the proliferation and migration of lung cancer cells, which is suppressed by LINC01635 silencing. (A) The knockdown efficiencies of miR-455-5p in A549 cell lines were examined by reverse transcription-quantitative PCR (*P<0.05). (B) CCK-8 assays showed that knockdown of miR-455-5p increased the proliferation of A549 cells (*P<0.05 and ***P<0.001 vs. NC inhibitor). (C) Transwell assays showed that knockdown of miR-455-5p increased the migration of A549 cells (*P<0.05). (D) CCK-8 assays showed that knockdown of miR-455-5p partially rescued the proliferation of A549 cells, which was suppressed by LINC01635 silencing (***P<0.001). (E) Transwell assays showed that knockdown of miR-455-5p partially rescued the migration of A549 cells, which was suppressed by LINC01635 silencing (***P<0.001). Scale bar, 100 µm. miR/miRNA, microRNA; si, small interfering; NC, negative control; LINC01635, long intergenic non-protein coding RNA 1635.
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References

    1. Bade BC, Dela Cruz CS. Lung cancer 2020: Epidemiology, etiology, and prevention. Clin Chest Med. 2020;41:1–24. doi: 10.1016/j.ccm.2019.10.001. - DOI - PubMed
    1. Venkatesan P. IASLC 2020 world conference on lung cancer. Lancet Respir Med. 2020;8:e76. doi: 10.1016/S2213-2600(20)30395-7. - DOI - PubMed
    1. Herbst RS, Morgensztern D, Boshoff C. The biology and management of non-small cell lung cancer. Nature. 2018;553:446–454. doi: 10.1038/nature25183. - DOI - PubMed
    1. Molina JR, Yang P, Cassivi SD, Schild SE, Adjei AA. Non-small cell lung cancer: Epidemiology, risk factors, treatment, and survivorship. Mayo Clin Proc. 2008;83:584–594. doi: 10.4065/83.5.584. - DOI - PMC - PubMed
    1. Hirsch FR, Scagliotti GV, Mulshine JL, Kwon R, Curran WJ, Jr, Wu YL, Paz-Ares L. Lung cancer: Current therapies and new targeted treatments. Lancet. 2017;389:299–311. doi: 10.1016/S0140-6736(16)30958-8. - DOI - PubMed

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The present study was supported by the Research Fund of Lianshui County People's Hospital (2020), and The Program of Innovation and Entrepreneurship Doctor of Jiangsu Province (2021).

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