LncRNA LOXL1-AS1 Research Articles

LOXL1-AS1 contributes to metastasis in sonic-hedgehog medulloblastoma by promoting cancer stem-like phenotypes

BackgroundMedulloblastomas (MBs) are one of the most common malignant brain tumor types in children. MB prognosis, despite improvement in recent years, still depends on clinical and biological risk factors. Metastasis is the leading cause of MB-related deaths, which highlights an unmet need for risk stratification and targeted therapy to improve clinical outcomes. Among the four molecular subgroups, sonic-hedgehog (SHH)-MB harbors clinical and genetic heterogeneity with a subset of high-risk cases. Recently, long non-coding (lnc)RNAs were implied to contribute to cancer malignant progression, but their role in MB remains unclear. This study aimed to identify pro-malignant lncRNAs that have prognostic and therapeutic significance in SHH-MB.MethodsThe Daoy SHH-MB cell line was engineered for ectopic expression of MYCN, a genetic signature of SHH-MB. MYCN-associated lncRNA genes were identified using RNA-sequencing data and were validated in SHH-MB cell lines, MB tissue samples, and patient cohort datasets. SHH-MB cells with genetic manipulation of the candidate lncRNA were evaluated for metastatic phenotypes in vitro, including cell migration, invasion, sphere formation, and expressions of stemness markers. An orthotopic xenograft mouse model was used to evaluate metastasis occurrence and survival. Finally, bioinformatic screening and in vitro assays were performed to explore downstream mechanisms.ResultsElevated lncRNA LOXL1-AS1 expression was identified in MYCN-expressing Daoy cells and MYCN-amplified SHH-MB tumors, and was significantly associated with lower survival in SHH-MB patients. Functionally, LOXL1-AS1 promoted SHH-MB cell migration and cancer stemness in vitro. In mice, MYCN-expressing Daoy cells exhibited a high metastatic rate and adverse effects on survival, both of which were suppressed under LOLX1-AS1 perturbation. Integrative bioinformatic analyses revealed associations of LOXL1-AS1 with processes of cancer stemness, cell differentiation, and the epithelial-mesenchymal transition. LOXL1-AS1 positively regulated the expression of transforming growth factor (TGF)-β2. Knockdown of TGF-β2 in SHH-MB cells significantly abrogated their LOXL1-AS1-mediated prometastatic functions.ConclusionsThis study proved the functional significance of LOXL1-AS1 in SHH-MB metastasis by its promotion of TGF-β2-mediated cancer stem-like phenotypes, providing both prognostic and therapeutic potentials for targeting SHH-MB metastasis.

A comprehensive review on lncRNA LOXL1-AS1: molecular mechanistic pathways of lncRNA LOXL1-AS1 in tumorigenicity of cancer cells.

Long non-coding RNAs (lncRNAs) are versatile RNAs that regulate various cellular processes, such as gene regulation, by acting as signals, decoys, guides, and scaffolds. A novel recognized lncRNA, LOXL1-antisense RNA 1 (LOXL1-AS1), is dysregulated in some diseases, including cancer, and acts as an oncogenic lncRNA in many types of cancer cells. Upregulation of LOXL1-AS1 has been involved in proliferation, migration, metastasis, and EMT, as well as inhibiting apoptosis in cancer cells. Most importantly, the malignant promoting activity of LOXL1-AS1 can be mostly mediated by sequestering specific miRNAs and inhibiting their binding to the 3´UTR of their target mRNAs, thereby indirectly regulating gene expression. Additionally, LOXL1-AS1 can decoy transcription factors and proteins and prevent their binding to their regulatory regions, inhibiting their mechanistic activity on the regulation of gene expression and signaling pathways. This review presents the mechanistic pathways of the oncogenic role of LOXL1-AS1 by modulating its target miRNAs and proteins in various cancer cells. Having information about the molecular mechanisms regulated by LOXL1-AS1 in cancer cells can open ways to find out particular prognostic biomarkers, as well as discover novel therapeutic approaches for different types of cancer.

LncRNA LOXL1-AS1 promotes proliferation and invasion and inhibits apoptosis in retinoblastoma by regulating the MAPK signaling pathway.

Retinoblastoma (RB) is an intraocular malignancy that is most common in children and rare in adults. Addressing novel biomarkers and therapeutic targets for RB to modulate tumor progression has become a challenge. The aim of the present study was to investigate the function of long non-coding RNAs (LncRNAs) LOXL1-AS1 in RB cell proliferation and metastasis. It was found that LOXL1-AS1 was overexpressed in RB tissues and cells. In order to evaluate cell viability and colony formation potential, the knockdown of LOXL1-AS1 has been established. Knockdown of LOXL1-AS1 was also inhibited cells migration and invasion. In addition, the proportion of cells in the G2/M phase of the sh-LOXL1-AS1 group increased significantly, and the proportion of cells in the sh-NC group decreased significantly. In the xenograft model of RB, the tumors in the sh-LOXL1-AS1 group grow slowly compared to the sh-NC group. Western blot analysis revealed that LOXL1-AS1 can regulate the progression of RB cells through MAPK signaling pathway in vitro and in vivo. These results indicated that LncRNA LOXL1-AS1 promotes proliferation, invasion and inhibits apoptosis of retinoblastoma by regulating MAPK signaling pathway, and might be expected to be a novel basis for clinical diagnosis and treatment.

Abstract 1285: The pro-metastatic role of long-noncoding RNA LOXL1-AS1 in sonic-hedgehog medulloblastoma

Abstract Introduction: Medulloblastoma (MB) is the most common malignant tumor in children. Since most MB-related deaths are the result of metastasis, current standard care for MB is aggressive with intense post-surgical radiotherapies, which aim to minimize metastatic occurences and are applied in most cases regardless of tumor subgroups. Among the four molecular subgroups, sonic-hedgehog (SHH-) MB has a high prevalence and well-characterized profiles. A proportion of SHH-MB cases with higher metastasis rate and worse prognosis have been linked to aberrant expressions of certain genes such as MYCN. Strategies to target such pro-metastatic drivers have been explored but proven to be challenging. Therefore, discovery of novel targets in MB is an urgent need to optimize treatment and improve outcomes. Long non-coding RNAs (lncRNAs) have recently emerged as malignancy regulators in many cancers. More insights into the role of lncRNAs in metastasis-promoting phenotypes, especially cancer stemness, would present both prognostic and therapeutic significance in metastatic MB. Objective: To determine the role of lncRNA LOXL1-AS1 in progression and metastasis of SHH-MB. Methods: First, we used RNA-sequencing to screen for differentially expressed lncRNAs in SHH-MB cells (Daoy) achieved a higher malignancy level via MYCN overexpression (Daoy-MYCN), identifying LOXL1-AS1 as one of the pro-malignant lncRNA candidates. LOXL1-AS1 expressions and prognostic associations were validated in clinical samples and datasets. Next, MB cells with stable LOXL1-AS1 knockdown (Daoy-MYCN) or overexpression (Daoy and ONS-76) were subjected to migration and invasion assays, as well as seeded in sphere-forming condition to assess sphere formation capacities and stemness marker expressions. Finally, we injected Daoy-MYCN cells knockdown of LOXL1-AS1 into BALB/c mouse brain to observe formation of tumor, metastasis, and survival rates. Results: LOXL1-AS1 was increased in Daoy cells under MYCN overexpression, was highly expressed in MB compared to normal brain, and was associated with poor prognosis in SHH-MB, particularly in the SHH-α and SHH-γ subgroups. MB cells with LOXL1-AS1 knockdown or overexpression showed a significant decrease or increase, respectively, in cell migration, cell invasion, sphere numbers, sphere diameters, and expression of stemness markers. Mice orthotopically transplanted with LOXL1-AS1-knockdown cells experienced a delayed tumor growth and metastasis development with improved overall and metastasis-free survivals. Conclusion: Our study has proven the role of LOXL1-AS1 in pro-metastatic phenotypes of MYCN-associated SHH-MB through a possible mechanism of inducing cancer stemness, which expands our current array of lncRNA candidates to develop MB-targeted therapies. The mechanisms of LOXL1-AS1-driven metastasis are being investigated in MB models. Citation Format: Anh Duy Do, Chia-Ling Hsieh, Tai-Tong Wong, Shian-Ying Sung. The pro-metastatic role of long-noncoding RNA LOXL1-AS1 in sonic-hedgehog medulloblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1285.

Molecular Mechanism of LncRNA LOXL1-AS1 Regulating the Proliferation and Apoptosis of Ovarian Cancer Cells by Targeting MicroRNA-761

Molecular Mechanism of LncRNA LOXL1-AS1 Regulating the Proliferation and Apoptosis of Ovarian Cancer Cells by Targeting MicroRNA-761

The LOXL1 antisense RNA 1 (LOXL1-AS1)/microRNA-423-5p (miR-423-5p)/ectodermal-neural cortex 1 (ENC1) axis promotes cervical cancer through the mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) pathway

ABSTRACT As the fourth commonest malignancy among females worldwide, cervical cancer (CC) poses a huge challenge to human health. The pivotal regulatory roles of lncRNAs in cancers have been highlighted. LOXL1 antisense RNA 1 (LOXL1-AS1) has been reported to play a key role in cervical squamous cell carcinoma and other various cancers. Thus, we investigated the roles and mechanisms of lncRNA LOXL1-AS1 in CC. The in vivo experiments demonstrated that LOXL1-AS1 downregulation inhibited tumor growth and metastasis and proliferation of CC cells. The results of RT-qPCR demonstrated that LOXL1-AS1 and ectodermal-neural cortex 1 (ENC1) expression levels were upregulated in CC cells and tissues, while microRNA-423-5p (miR-423-5p) level was downregulated. As subcellular fractionation assays, RNA pull down assays and luciferase reporter assays revealed, LOXL1-AS1 bound to miR-423-5p and miR-423-5p targeted ENC1. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, wound healing and colony formation assays demonstrated that miR-423-5p upregulation and LOXL1-AS1 downregulation inhibited CC cell proliferation and migration, while ENC1 upregulation attenuated the inhibitory effects of miR-423-5p upregulation on the malignant phenotypes of CC cells. Western blotting was conducted to measure protein levels and the results showed that ENC1 knockdown inhibited the activation of ERK/MEK pathway. In summary, the LOXL1-AS1/miR-423-5p/ENC1 axis accelerates CC development through the MEK/ERK pathway.

LncRNA LOXL1-AS1 Facilitates the Oncogenic Character in Cervical Cancer by the miR-526b-5p /LYPLA1 Axis.

Increasing reports demonstrate that long noncoding RNAs participate in the regulation of numerous malignancies, cervical cancer included. Although lncRNA LOXL1 antisense RNA 1 has been commonly accepted to be an oncogene in many cancers. Here, the role of LOXL1-AS1 in CC still need to be explored. In this study, LOXL1-AS1 was found elevated in CC tissues and cells. LOXL1-AS1 depletion restrained CC cell proliferation, migration, invasion, and angiogenesis in vivo. Furthermore, we found that LOXL1-AS1 upregulated Lysophospholipase 1 expression via sequestering miR-526b-5p. Rescue assays revealed that overexpression of LYPLA1 reversed the LOXL1-AS1 silencing-induced inhibitory effects on the malignant phenotypes of CC cells. To conclude, this study showed that LOXL1-AS1 facilitates cellular process in CC via functioning as a miR-526b-5p sponge.

LncRNA LOXL1-AS1 inhibits proliferation of PDLSCs and downregulates IL-1β in periodontitis patients.

The carcinogenic role of lncRNA LOXL1-AS1 in thoracic aortic aneurysm has been confirmed, but its role in the pathogenesis of periodontitis remains unclear. Our preliminary deep sequencing data revealed LOXL1-AS1 downregulation in periodontitis and its inverse correlation with IL-1β, a critical inflammatory mediator in periodontitis. This study was therefore performed to investigate the potential interaction between LOXL1-AS1 and IL-1β in periodontitis. The study included 30 periodontitis patients (18males and 12 females at the age of 34 to 44years, with a mean of 39.3±2.1years) and 30healthy controls (18males and 12 females at the age of 33 to 44years with a mean of 39.2±2.0years). The effects of LOXL1-AS1 overexpression on IL-1β were evaluated by RT-qPCR and Western blot. CCK-8 assay was used to analyze cell proliferation. LOXL1-AS1 was downregulated in periodontitis-affected periodontal ligament stem cells (PDLSCs) compared with healthy PDLSCs, while IL-1β was upregulated in periodontitis-affected PDLSCs and was inversely correlated with LOXL1-AS1. LOXL1-AS1 overexpression mediated IL-1β downregulation in PDLSCs. IL-1β treatment did not affect LOXL1-AS1 expression. Moreover, LOXL1-AS1 overexpression inhibited the proliferation of periodontitis-affected PDLSCs. LOXL1-AS1 overexpression and IL-1β knockdown increased Bax/Bcl-2 ratio and caspase-3level. This study is the first to report LOXL1-AS1 downregulation in periodontitis. Moreover, LOXL1-AS1might inhibit the proliferation of periodontitis-affected PDLSCs and downregulate IL-1β to improve periodontitis.

LOXL1‑AS1 promotes thymoma and thymic carcinoma progression by regulating miR‑525‑5p‑HSPA9.

Due to the lack of specific symptoms in early thymic epithelial tumours (TETs), patients are mostly in the advanced stage at the time of presentation. The aim of the present study was to explore the mechanism by which the long noncoding RNA (lncRNA) LOXL1-AS1 affects thymoma and thymic carcinoma progression by targeting the miR-525-5p-HSPA9 axis. Bioinformatics was used to analyse the process of LOXL1-AS1 targeting miR-525-5p-HSPA9 and its expression characteristics in TET. The relationships between LOXL1-AS1, miR-525-5p, HSPA9 and prognosis were analysed. The dual luciferase reporter assay was applied to verify targeting. The gene was knocked down or overexpressed by plasmid transfection. Cell counting kit 8 (CCK-8) assay, flow cytometry and Transwell assay were used to detect cell viability, apoptosis and invasion ability, respectively. Proteins and RNAs were examined by western blot analysis and qPCR, respectively. A tumour-burdened assay was used to perform in vivo verification. LOXL1-AS1 and HSPA9 were overexpressed in thymoma and thymic carcinoma; high levels of LOXL1-AS1 and HSPA9 were associated with poor prognosis, and there was a significant positive correlation between their levels. Downregulation of miR-525-5p expression was also associated with poor prognosis of patients. Clinical trials also demonstrated the same trends. miR-525-5p inhibited the expression of HSPA9 protein by targeting the 3′-untranslated region (UTR) of HSPA9 mRNA. LOXL1-AS1 promoted the expression of HSPA9 as a sponge targeting miR-525-5p. Animal experiment results also showed that knockdown of miR-525-5p promoted cancer by promoting the expression of HSPA9. In conclusion, LOXL1-AS1 and HSPA9 are highly expressed in thymoma and thymic carcinoma; miR-525-5p is expressed at low levels in thymoma and thymic carcinoma; and downregulation of miR-525-5p is associated with poor prognosis. In summary, this study demonstrates that LOXL1-AS1 acts as a sponge that targets miR-525-5p to promote HSPA9 expression, thereby promoting the growth and invasion and inhibiting apoptosis of thymoma and thymic carcinoma cells.

LncRNA LOXL1-AS1 promotes esophageal squamous cell carcinoma progression by targeting DESC1.

Recently, ample evidence indicated that numerous aberrantly expressed long non-coding RNAs (lncRNAs) participated in the development of multiple malignancies. However, the expression and function of lncRNA LOXL1-AS1 in mediating esophageal squamous cell carcinoma (ESCC) carcinogenesis remains largely elusive. Here we validated that LOXL1-AS1 was significantly upregulated in ESCC tissues compared with the corresponding adjacent non-neoplastic tissues, and LOXL1-AS1 expression was positively correlated with ESCC patients' lymph node metastasis. Besides, LOXL1-AS1 knockdown impaired ESCC cells proliferation, migration and invasion capabilities in vitro. Furthermore, inhibiting LOXL1-AS1 in ESCC cells increased the percentage of cells at the G1 phase, accompanied by reducing in S phase in contrast to scramble control, and silencing of LOXL1-AS1 evoked ESCC cell apoptosis. From high throughput RNA sequencing (RNA-seq) analysis, we identified that differentially expressed in squamous cell carcinoma 1 (DESC1) was a critical downstream target of LOXL1-AS1. Taken together, we demonstrated the function and mechanism of LOXL1-AS1 in contributing ESCC progression for the first time, and indicated LOXL1-AS1 may be a novel therapeutic biomarker of ESCC.

Targeted-regulating of miR-515-5p by LncRNA LOXL1-AS1 on the proliferation and migration of trophoblast cells

ObjectiveThis study aims to elucidate the molecular mechanism of LOXL1-AS1 in proliferation and migration of trophoblast cells. MethodsWe have used specific small interfering RNAs (si-RNA) and microRNA (mi-RNA) to knock down the target gene or m-RNA. In this regard we used following siRNAs: si-NC, si-LOXL1-AS1, pcDNA-NC, pcDNA-LOXL1-AS1, miR-NC, miR-515-5p. These si-RNA and miRNA were transfected into JeG-3 cells. Real-time quantitative PCR (RT-qPCR) was used to detect the expressions of LOXL1-AS1 and miR-515-5p. Western blot was used to detect Cyclin D1, matrix metalloproteinase 2 (MMP2), matrix metalloproteinases 9 (MMP9), phosphorylated p65 (p-p65), profilin of phosphorylated nuclear transcription factor κB (p-IκBα). MTT assay was used to detect cell survival rate, and the luciferase assay was used to detected the targeting relationship of LOXL1-AS1 and miR-515-5p. ResultsOur results showed that human placental trophoblast cells had higher level of LOXL1-AS1 in comparison to human choriocarcinoma cells, however, human placental trophoblast cells had lower level of miR-515-5p. In addition, the expression of CyclinD1, MMP2, MMP9 were significantly decreased in JeG-3 cell lines. We observed lower cell survival rate and lower cell migration number in JeG-3 cell lines. Our results demonstrated that LOXL1-AS1 could target miR-515-5p, and subsequently reverse the inhibitory effect of LOXL1-AS1 on proliferation and migration in JEG-3 cells. Also, lower expressions of p-p65 and p-IкBα in JeG-3 cells showed that miR-515-5p could reversed the inhibitory effect of LOXL1-AS1 on NF-κB signaling pathway. ConclusionsThe low expression of LOXL1-AS1 inhibits the proliferation and migration of human choriocarcinoma cells, which might be related to miR-515-5p and NF-κB signaling pathways.

Up-regulation of long non-coding RNA LOXL1-AS1 functions as an oncogene in cervical squamous cell carcinoma by sponging miR-21

LncRNA LOXL1-AS1 has been reported to be upregulated in several types of cancer and plays oncogenic roles. We analysed TCGA dataset and observed the downregulation of LOXL1-AS1 in cervical squamous cell carcinoma (CSCC). We therefore analysed the roles of LOXL1-AS1 in CSCC. We observed that LOXL1-AS1 was downregulated in CSCC. LOXL1-AS1 was predicted to interact with miR-21, while overexpression experiments showed that LOXL1-AS1 and miR-21 had no significant effects on the expression of each other. However, LOXL1-AS1 overexpression led to the upregulation of RHOB, a direct target of miR-21. Cell invasion and migration analysis showed decreased invasion and migration rates of CSCC cells after LOXL1-AS1 and RHOB overexpression. MiR-21 played an opposite role at reduced the effects of LOXL1-AS1 and RHOB overexpression. Therefore, LOXL1-AS1 may promote CSCC cell invasion and migration by sponging miR-21 to upregulate RHOB.

Long Non-Coding RNA LOXL1-AS1 Enhances Colorectal Cancer Proliferation, Migration and Invasion Through miR-708-5p/CD44-EGFR Axis.

IntroductionColorectal cancer (CRC), the third most common cancer worldwide, involves a physiological and pathological long non-coding RNA (lncRNA) paradigm shift. It has been reported that the lncRNA LOXL1-AS1 affects tumor development for many kinds of cancers, but its functions and mechanisms in CRC remain unknown.MethodsExpression levels of LOXL1-AS1 and miR-708-5p within CRC tissues and cell lines were measured using qRT-PCR. The performance of gain-of-function and loss-of-function assays was aimed at examining the effects of LOXL1-AS1 and miR-708-5p; colony formation and cell viability assays were carried out to measure cell multiplication; and Transwell migration and wound-healing assays were carried out for the measurement of cell migration and invasion. Luciferase reporter assay was used to verify the interactions between LOXL1-AS1 and miR-708-5p and between miR-708-5p and the CD44-EGFR signaling pathway. Finally, expression of CD44 and EGFR proteins was measured by Western blot and immunofluorescence assays.ResultsIn this study, we reveal that the regulation of lncRNA LOXL1-AS1 occurs within CRC based on the correlation with poor clinical outcomes. LOXL1-AS1 knockdown along with miR-708-5p overpresentation in CRC cell lines inhibited cell multiplication, migration, and invasion. The inhibiting effect of LOXL1-AS1 knockdown on CRC was reversed by upregulating the CD44-EGFR signal pathway. From the perspective of mechanism, LOXL1-AS1 imposes sponging upon miR-708-5p and thereby promotes the CD44-EGFR signal pathway in CRC cells.DiscussionThis study demonstrated that lncRNA LOXL1-AS1 enhances multiplication, migration, invasion, and progression of CRC by sponging miR-708-5p to regulate the CD44-EGFR signal pathway.

LncRNA LOXL1-AS1 controls osteogenic and adipocytic differentiation of bone marrow mesenchymal stem cells in postmenopausal osteoporosis through regulating the miR-196a-5p/Hmga2 axis.

Exploring molecular mechanisms of human bone marrow mesenchymal stem cells (hBMMSCs) differentiation, a crucial step for bone formation, is a new direction for treating postmenopausal osteoporosis. LncRNAs are involved in lots of biological processes including hBMMSCs differentiation. The present study aimed to explore the effect of LOXL1-AS1 on hBMMSCs differentiation. We examined the expression levels of LOXL1-AS1, miR-196a-5p and Hmga2 in peripheral blood from postmenopausal osteoporosis patients by RT-qPCR, and detected their changes during the osteogenic differentiation of hBMMSCs by RT-qPCR. RT-qPCR and western blot measured the level of biomarkers of bone formation and osteogenic differentiation (osteopontin, OPN; Alkaline phosphatase, ALP; Runt-related transcription factor-2, Runx-2). The effects of LOXL1-AS1 on the osteogenic and adipocytic differentiation of hBMMSCs were, respectively, determined by ALP, ARS staining assays and oil red O staining assay. The abnormal high expression of LOXL1-AS1 was found in patients. LOXL1-AS1 expression showed a gradual decrease during the osteogenic differentiation of hBMMSCs. However, LOXL1-AS1 overexpression inhibited the hBMMSCs osteogenic differentiation but promoted adipocytic differentiation. Furthermore, LOXL1-AS1 was identified to be a sponge of miR-196a-5p and Hmga2 as a target gene of miR-196a-5p. Besides, LOXL1-AS1 sponged miR-196a-5p to mediate Hmga2 expression, which played contrary effects on regulating osteogenic and adipocytic differentiation of hBMMSCs. Moreover, LOXL1-AS1/miR-196a-5p/Hmga2 axis regulated hBMMSCs differentiation through controlling C/EBPβ-mediated PPARγ expression. These findings facilitate understanding the molecular mechanism of hBMMSCs differentiation and bring up a novel sight for postmenopausal osteoporosis therapy.

LOXL1-AS1 Contributes to Non-Small Cell Lung Cancer Progression by Regulating miR-3128/RHOXF2 Axis.

PurposeThe purpose of this study was to investigate the molecular mechanism of LncRNA LOXL1-AS1 in non-small cell lung cancer (NSCLC).MethodsLung cancer cell lines (H1299, A549, H520 and H596) and human normal lung epithelial cell line (BEAS-2B) were used in this study. Gene expression was measured by qRT-PCR (quantitative real-time PCR). The bioinformatics databases (miRDB and TargetScan7) were used to predict target genes. Luciferase assay and pull-down assay were processed for verifying the binding sites. CCK8 assay was used for detecting proliferation, and transwell assay was undertaken for migration and invasion.ResultsLncRNA LOXL1-AS1 was higher expressed in lung cancer tissues and cells. Moreover, LOXL1-AS1 expression was upregulated in tumor tissues with advanced stages and metastasis. After knocking down LOXL1-AS1, proliferation, invasion and migration of H1299 and A549 cells were inhibited. Interestingly, miR-3128 was negatively regulated by LncRNA LOXL1-AS1, which inhibited the expression of RHOXF2. Rescue assay also confirmed that miR-3128 inhibitor and oeRHOXF2 could rescue the effect of down-regulated LOXL1-AS1 on proliferation, invasion and migration progression.ConclusionLOXL1-AS1 promotes the progression of NSCLC by regulating miR-3128/RHOXF2 axis, which might be a new potential target for the diagnosis and treatment of NSCLC.

Non‐coding RNA LOXL1-AS1 exhibits oncogenic activity in ovarian cancer via regulation of miR‐18b‐5p/VMA21 axis

Long non-coding RNAs (lncRNAs) exert critical effects in the process of malignant cancers and lncRNA LOXL1 Antisense RNA 1 (LOXL1-AS1) has been demonstrated to be a pro-oncogene in multiple tumor types. In the current study, we illuminated the precise roles of LOXL1-AS1 in the development of ovarian cancer. LOXL1-AS1 is significantly overexpressed in ovarian carcinoma tissue compared with adjacent non‐cancerous sample. The luciferase reporter gene assay reveals the relationship between LOXL1-AS1 and miR‐18b‐5p, miR‐18b‐5p and its target gene, Vacuolar ATPase Assembly Factor VMA21 (VMA21). Transfection of LOXL1-AS1 siRNA or miR‐18b‐5p mimics inhibits the growth and aggressive phenotypes of SKOV3 and OVCAR3 cell. Furthermore, miR-18b-5p suppresses ovarian carcinoma cell proliferation and metastasis by targeting VMA21 and LOXL1-AS1 regulates ovarian carcinoma cell growth and metastasis through sponging miR‐18b‐5p. These findings suggest that lncRNA LOXL1-AS1 promotes ovarian cancer cell growth, migratory and invasiveness via modulating miR‐18b‐5p/VMA21 axis.

LncRNA LOXL1-AS1 promotes endometrial cancer progression by sponging miR-28-5p to upregulate RAP1B expression

BackgroundIncreasing lncRNAs are found to be involved in the biological process of multiple cancer types. Herein, we aimed to reveal the role of LOXL1-AS1 in endometrial cancer (EC) progression. MethodsTumor and corresponding normal tissues were obtained from EC patients. Si-LOXL1-AS1 and miR-28-5p inhibitor were transfected to downregulate the expressions of LOXL1-AS1 and miR-28-5p, while miR-28-5p mimics were used to upregulate the miR-28-5p expression. CCK-8 and colony assays were applied to estimate the cell proliferation. Flow cytometry was performed to measure the cell apoptosis. Wound healing and transwell assays were conducted to assess the cell migration and invasion abilities. Informatics analysis was used to explore the relationship among LOXL1-AS1, miR-28-5p and RAP1B. ResultsLOXL1-AS1 was found markedly up-regulated in EC tissues and cell lines. LOXL1-AS1 knockdown displayed evident suppression in cell proliferation, migration and invasion, as well as promotion in cell apoptosis. Moreover, the LOXL1-AS1 induced regulatory effects on EC cells were partially reversed by miR-28-5p inhibitor. Mechanistically, LOXL1-AS1 competitively bond to miR-28-5p, resulting in upregulation of RAP1B. Additionally, in vivo study confirmed the findings discovered in vitro. ConclusionsIn summary, LOXL1-AS1 exerted oncogenic roles in EC progression by sponging miR-28-5p and thereby upregulating RAP1B. This finding might provide potential targets for EC therapy.

LOXL1-AS1 Drives The Progression Of Gastric Cancer Via Regulating miR-142-5p/PIK3CA Axis.

BackgroundGastric cancer (GC) is a deadly disease, and its incidence is especially high in East Asia including China. Recently, some long non-coding RNA (lncRNAs) have been identified as oncogenes or tumor suppressors. This study aimed to determine the function and mechanism of lncRNA LOXL1-AS1 on the progression of GC.MethodsRT-PCR was done to measure the expression levels of LOXL1-AS1 and miR-142-5p in GC tissues. The association between pathological indexes and LOXL1-AS1 expression was also analyzed. Human GC cell lines AGS and BGC823 were used as cell models. CCK-8 and colony formation assays were conducted to assess the effect of LOXL1-AS1 on the proliferation of GC cell lines. Transwell assay was conducted to determine the influence of LOXL1-AS1 on cell migration and invasion. Furthermore, luciferase reporter assay was carried out to confirm the relationship of miR-142-5p with LOXL1-AS1. Additionally, Western blot was done to detect the regulatory function of LOXL1-AS1 on PIK3CA, a target of miR-142-5p. In vivo experiment was also performed to validate the roles and mechanism of LOXL1-AS1 on the growth and metastasis of GC cells.ResultsLOXL1-AS1 expression in GC samples was significantly increased, which was correlated with unfavorable pathological indexes. Highly expressed LOXL1-AS1 was closely linked to shorter overall survival time and post-progression survival time of the patients. LOXL1-AS1 markedly modulated the malignant phenotypes of GC cells. Additionally, overexpressed LOXL1-AS1 notably reduced the expression of miR-142-5p, but enhanced the expression level of PIK3CA. In vivo experiments further validated that knockdown of LOXL1-AS1 inhibited the growth and metastasis of GC cells via regulating miR-142-5p and PIK3CA.ConclusionLOXL1-AS1 was a sponge of tumor suppressor miR-142-5p in GC, enhanced the expression of PIK3CA indirectly and functioned as an oncogenic lncRNA.

LncRNA LOXL1-AS1 inhibited cell proliferation, migration and invasion as well as induced apoptosis in breast cancer via regulating miR-143-3p.

In recent years, studies have shown that lncRNA plays an essential regulatory role in biological life activities. In disease and cancer research, the function of lncRNA is closely related to inflammatory response, tumor formation and cellular metabolism. Breast cancer is one of the most common malignant tumors in women. The research on the pathogenesis of breast cancer is the focus of current research. Although the regulatory mechanisms of some lncRNAs have been proven, the complexity of cancer regulation has led to incomplete research. The expression of LOXL1-AS1 and miR-143-3p was measured using qRT-PCR. Western blot was used to detect CDK, Cyclin D1, MMP-2, MMP-9, Bcl-2, Bax and Cleaved caspase-3 protein expression. MTT assay and transwell assay were applied to analyze cell proliferation, migration and invasion, respectively. Cell apoptosis rate of transfected cells was measured with flow cytometry. Luciferase reporter assay was applied to verify the relationship between LOXL1-AS1 and miR-143-3p. In this study, we found that LOXL1-AS1 expression was induced while miR-142-3p expression was decreased in breast cancer tissues and cells, implying that LOXL1-AS1 and miR-143-3p play an important role in cell progression of breast cancer. Further investigation showed that silencing LOXL1-AS1 inhibited proliferation, promoted cell apoptosis and decreased the capacity of cell migrated and invasive in breast cancer cells. The analysis of luciferase reporter assay determined that LOXL1-AS1 directly targeted miR-143-3p in breast cancer cells. The rescue experiments further proved that miR-143-3p reversed the inhibited effects of si- LOXL1-AS1 on breast cancer cells. In this study, we verified that LncRNA LOXL1-AS1 inhibited cell proliferation, migration and invasion as well as induced apoptosis in breast cancer via regulating miR-143-3p, providing a novel therapeutic target and improving understanding of the regulatory mechanism of cell progression in breast cancer.

LncRNA LOXL1-AS1 regulates the tumorigenesis and development of lung adenocarcinoma through sponging miR-423-5p and targeting MYBL2.

Lung adenocarcinoma (LUAD) is the most common form of malignant tumor and closely correlated with high risk of death worldwide. Accumulating researches have manifested that long noncoding RNAs (lncRNAs) are deeply involved in the progression of multiple cancers. LncRNA LOXL1 antisense RNA 1 (LOXL1‐AS1) was identified as an oncogene in several cancers, nonetheless, its biological effect and regulatory mechanism have not been explained in LUAD. Our present study suggested that LOXL1‐AS1 expression was considerably increased in LUAD tissues and cells. Moreover, LOXL1‐AS1 deficiency notably hampered cell proliferation and migration as well as dramatically facilitated cell apoptosis. Through molecular mechanism assays, LOXL1‐AS1 was identified as a cytoplasmic RNA and acted as a sponge of miR‐423‐5p. Furthermore, MYBL2 was targeted and negatively modified by miR‐423‐5p. Rescue experiments revealed that MYBL2 knockdown could counteract miR‐423‐5p repression‐mediated enhancement on the progression of LOXL1‐AS1 downregulated LUAD cells. More importantly, MYBL2 was discovered to interact with LOXL1‐AS1 promoter, indicating a positive feedback loop of LOXL1‐AS1/miR‐423‐5p/MYBL2 in LUAD. These findings manifested the carcinogenic role of LOXL1‐AS1 and LOXL1‐AS1/miR‐423‐5p/MYBL2 feedback loop in LUAD, which could be helpful to explore effective therapeutic strategy for LUAD patients.