Ultrasound-targeted microbubble destruction mediated miR-492 inhibitor suppresses the tumorigenesis in non-small cell lung cancer

BackgroundUltrasound-targeted microbubble destruction (UTMD) is a novel adjuvant tumor therapeutic method by enhancing exogenous gene transfection to target tissues. This study aims to investigate the role of microRNA-492 (miR-492) in non-small cell lung cancer (NSCLC) and further analyze the effects of UTMD-mediated miR-492 inhibitor on tumorigenesis.MethodsThe expression of miR-492 was detected by qRT-PCR. Co-transfection of microbubbles and miR-492 inhibitor with Lipofectamine 3000 was performed to achieve UTMD-mediated miR-492 inhibition in NSCLC cells. CCK-8 and Transwell assay were used to determine NSCLC cell proliferation, and the migration and invasion.ResultHigh expression of miR-492 was associated with poor prognosis in NSCLC patients. miR-492 inhibitor suppressed tumor cell proliferation, migration and invasion, and UTMD not only increased the transfection efficiency of miR-492 inhibitor, but also enhance the inhibitory effects on cell biological behaviors.ConclusionThe results showed that the expression level of miR-492 was up-regulated in NSCLC tissue samples and cells. Silencing of miR-492 inhibited NSCLC cell proliferation, migration and invasion, and UTMD-mediated miR-492 inhibitor could promote more significant inhibition, which indicated that UTMD-mediated miR-492 inhibitor might provide a novel strategy for the treatment of NSCLC.

KEY MESSAGES

• miR-492 inhibitor inhibited cell proliferation, migration and invasion.
• UTMD-mediated miR-492 inhibitor can promote more significant inhibition.
• UTMD-mediated miR-492 inhibitor provide a new strategy for NSCLC.

     

SPOCD1 regulated by miR-133a-3p promotes hepatocellular carcinoma invasion and metastasis

ObjectiveTo investigate the tumorigenic role of spen paralogue and orthologue C-terminal domain-containing 1 (SPOCD1) in hepatocellular carcinoma (HCC) and identify the upstream regulatory mechanism.MethodsWe analyzed SPOCD1 and miR-133-3p expression in normal and HCC tissues from the Cancer Genome Atlas and UALCAN databases, and in normal hepatocytes and HCC cell lines by real-time quantitative polymerase chain reaction and western blot. We identified the miR-133a-3p-binding site on the SPOCD1 3ʹ-untranslated region using TargetScan. Hierarchical regulation was confirmed by luciferase assay and miR-133a-3p overexpression/silencing. Cell proliferation, migration, invasion, and colony formation were assessed by MTT, scratch, transwell, and clonogenic assays, respectively.ResultsSPOCD1 was highly expressed in HCC tissues and cell lines, while miR-133a-3p expression was significantly downregulated. Kaplan–Meier analysis indicated that high SPOCD1 expression was significantly associated with poor survival. TargetScan and luciferase reporter assay revealed that SPOCD1 was the downstream target of miR-133a-3p. Overexpression of miR-133a-3p significantly inhibited the expression of SPOCD1, while miR-133a-3p knockdown significantly increased SPOCD1 expression.ConclusionSPOCD1, regulated by miR-133a-3p, promotes HCC cell proliferation, migration, invasion, and colony formation. This study provides the first evidence for the role of the miR-133a-3p/SPOCD1 axis in HCC tumorigenesis.     

MiR-206 inhibits Head and neck squamous cell carcinoma cell progression by targeting HDAC6 via PTEN/AKT/mTOR pathway

As a kind of endogenous noncoding small RNA, microRNA (miRNA) plays important roles of regulation to various physiological functions, while its affections on senescence of human Head and neck squamous cell carcinoma (HNSCC) are still unknown. The objective of this study was to investigate the effect of miR-206 in HNSCC tissues, adjacent normal tissues and cell lines, and explore its biological functions in HNSCC.In our study, the level of miR-206 in HNSCC tissues and adjacent normal tissues was detected via real-time qPCR. The effect of miR-206 on cell proliferation was tested by MTT assay, colony formation and cell cycle assays. In order to explore the effect of miR-206 on HNSCC cell migration and invasion, we performed wound healing assays and transwell assays. Luciferase reporter assays were designed to identify the interaction between 3′UTR of HDAC6 and miR-206. The level of signaling pathway-related proteins was determined by western blot. The expression of miR-206 was found to be observably decreased in HNSCC tissues and cell lines through real time-PCR. Restoration of miR-206 weaked cell proliferation, invasion and migration in HNSCC cells and the cell cycle was arrest in S phase. Further explores have shown that miR-206 could inhibit HNSCC cells proliferation by targeting the HDAC6 via PTEN/AKT/mTOR pathway. Taken together, our results demonstrated that miR-206 plays a critical role in HNSCC progression by targeting HDAC6 via PTEN/AKT/mTOR pathway, which might be a potential target for diagnostic and therapeutic in HNSCC.     

Hsa_circ_0042823 accelerates cancer progression via miR-877-5p/FOXM1 axis in laryngeal squamous cell carcinoma

BackgroundLaryngeal squamous cell carcinoma (LSCC) is a common malignant tumour of the head and neck. Our previous study reveals that the circular RNA (circRNA) hsa_circ_0042823 is abnormally expressed in LSCC, suggesting that hsa_circ_0042823 is closely associated with LSCC. Here, we attempted to explore the molecular mechanism of hsa_circ_0042823 in LSCC.MethodsQuantitative real-time PCR and western blot were performed to assess the expression of gene and protein in human laryngeal carcinoma cells, TU212 and TU686. MTT and transwell assays were performed to examine cell proliferation, migration and invasion. The relationship among hsa_circ_0042823, miR-877-5p and forkhead box M1 (FOXM1) was verified by luciferase reporter assay. Finally, we constructed a subcutaneous tumour mouse model to analyse in vivo growth of LSCC cells following knockdown of hsa_circ_0042823.ResultsCompared with normal human bronchial epithelial cells (HBECs), hsa_circ_0042823 was highly expressed in the LSCC cell lines (AMC-HN-8 and TU686). Further studies demonstrated that hsa_circ_0042823 interacted with miR-877-5p, and FOXM1 was the target of miR-877-5p. Hsa_circ_0042823 promoted the expression of FOXM1 via its ceRNA activity on miR-877-5p. Hsa_circ_0042823 overexpression promoted proliferation, migration and invasion of AMC-HN-8 cells through regulating miR-877-5p/FOXM1 axis. Additionally, inhibition of hsa_circ_0042823 inhibited growth of LSCC in vivo via miR-877-5p/FOXM1 axis.ConclusionsHsa_circ_0042823/miR-877-5p/FOXM1 axis participates in the progression of LSCC. This work demonstrates that hsa_circ_0042823 accelerates cancer progression by regulating miR-877-5p/FOXM1 axis in LSCC. Therefore, this study may provide new insights into the pathogenesis of LSCC.

KEY MESSAGES

• Hsa_circ_0042823 promotes FOXM1 expression by sponging miR-877-5p.
• Hsa_circ_0042823 promotes proliferation, migration, invasion of LSCC cells.
• Hsa_circ_0042823 knockdown inhibits tumour growth of LSCC via miR-877-5p/FOXM1 axis.

     

Retracted Article: MiR-206 reduced the malignancy of hepatocellular carcinoma cells in vitro by inhibiting MET and CTNNB1 gene expressions

The anti-cancer role of miR-206 in hepatocellular carcinoma (HCC) cells has been reported, but its mechanism of action remains poorly understood. This research aimed to investigate the anti-HCC mechanism of miR-206. We analyzed 25 pairs of HCC and adjacent tissue specimens from HCC patients. Two patient-derived HCC cell lines were established. MiR-206 levels in tissue specimens and cell lines were detected by qRT-PCR. MiR-206 overexpression was mimicked by miR-206 mimic transfection. MET or CTNNB1 gene was overexpressed by transient transfection. Protein and protein phosphorylation levels of interests were assessed by western blotting. HCC cell malignancy in vitro was evaluated by cell proliferation, apoptosis, colony formation, trans-well invasion assays as well as western blotting assessing the marker proteins of epithelial or mesenchymal phenotype. We found that miR-206 level was significantly lower in HCC tissue specimens in comparison to adjacent counterparts. Two patient-derived HCC cell lines showed lower miR-206 level than L02 human hepatocytes. MiR-206 mimic transfection significantly reduced phosphorylation levels of pan-Akt Ser9, Erk1 Thr202/Tyr204 and Gsk-3beta Ser308 as well as protein levels of beta-catenin and c-Met in primary HCC cells in vitro. Luciferase reported assay and AGO2-RNA co-immunoprecipitation assays results demonstrated that miR-206 reduced MET and CTNNB1 gene expressions in HCC cells by interacting with the 3′ UTR of their mRNAs. Restoring c-Met or beta-catenin protein level by MET or CTNNB1 transient overexpression partially restored the malignancy of HCC cells in vitro. We concluded that miR-206 might inhibit HCC development by targeting MET and CTNNB1 gene expression.

The anti-cancer role of miR-206 in hepatocellular carcinoma (HCC) cells has been reported, but its mechanism of action remains poorly understood.     

Effect of MicroRNA-218 on the viability,apoptosis and invasion of renal cell carcinoma cells under hypoxia by targeted downregulation of CXCR7 expression

ObjectiveTo investigate the effect of microRNA-218 on the viability, apoptosis and invasion of renal cell carcinoma cells under hypoxia by targeted regulation of expression of chemokine receptor 7 (CXCR7).MethodsThe expression of miR-218 in renal cell carcinoma cell lines under normal and hypoxia conditions, as well in normal renal tubular epithelial cells (HK2) was measured using RT-PCR. MiR-218 mimic and NC were transfected into renal cell carcinoma cell line ACHN using Lipofectamine™ 2000. The expression of miR-218 was analyzed using RT-PCR. The viability, apoptosis, migration and invasion of the transfected cells were assayed using the MTT assay, flow cytometry and transwell assays. The expression of CXCR7 was assayed using RT-PCR and Western blot. Luciferase reporter was used to verify the downstream target of miR-218.ResultsThe expression of miR-218 was lower than in renal cell carcinoma cell lines ACHN, 769-p and Caki-1 that in HK-2. The expression of miR-218 in the renal carcinoma cell lines was lower under hypoxia than under normal oxygen conditions. The expression of miR-218 in ACHN cells under normal and hypoxic conditions was significantly increased after transfection with miR-218 mimic. Compared with NC transfected cells under normal oxygen condition, the mimic-transfected cells had reduced viability, migration ability and invasion ability, and increased apoptosis, and mimic transfected-cells under hypoxia had significantly reduced viability, migration ability and invasion ability, and increased apoptosis. Overexpression of miR-218 mimic resulted in significant reduction in the expression of CXCR7 at protein and mRNA levels under normal and hypoxic conditions. Luciferase reporter assay confirmed that CXCR7 is the target protein of miR-218.ConclusionUp-regulation of miR-218 expression in renal cell carcinoma under hypoxia can result in significant and targeted down-regulation of CXCR7 expression, which could reduce cell viability, migration and invasion ability and induce apoptosis in the cancer cells.     

LINC01232 serves as a novel biomarker and promotes tumour progression by sponging miR-204-5p and upregulating RAB22A in clear cell renal cell carcinoma

BackgroundLong non-coding RNAs (lncRNAs) are involved in the progression of various cancers, including clear cell renal cell carcinoma (ccRCC). This study aimed to investigate the expression and prognostic value of long intergenic non-protein coding RNA (LINC) 01232 in ccRCC and preliminary explore the molecular mechanism underlying the role of LINC01232 in ccRCC progression.MethodsTumour tissues and adjacent normal tissues of 122 patients with ccRCC were collected in this study. The levels of LINC01232, microRNA (miR)-204-5p and RAB22A were measured by quantitative real-time PCR. The proliferation, migration and invasion of ccRCC cells were detected by cell counting kit-8 assay and Transwell assay, respectively. The interaction among LINC01232, miR-204-5p and RAB22A was confirmed by bioinformatics analysis, dual-luciferase reporter assay and Pearson correlation analysis. The association of LINC01232 and miR-204-5p with ccRCC patient survival was verified by the Kaplan–Meier method and log-rank test. The prognostic value of LINC01232 in ccRCC was confirmed by Cox regression analysis.ResultsLINC01232 expression was increased in ccRCC tumour tissues and ccRCC cells and independently predicted the prognosis of ccRCC patients. In addition, LINC01232 silencing inhibited ccRCC cell proliferation, migration and invasion. Moreover, LINC01232 served as a sponge for miR-204-5p, and miR-204-5p reduction reversed the inhibitory effect of LINC01232 silencing on ccRCC cell function. Furthermore, LINC01232 could sponge miR-204-5p, causing the elevation of RAB22A in ccRCC, thereby promoting ccRCC cell function.ConclusionLINC01232 may be an independent prognostic biomarker in ccRCC and plays an oncogenic role in ccRCC progression by sponging miR-204-5p and upregulating RAB22A.     

miR-301-3p directly regulates Cx43 to mediate the development of gastric cancer

ObjectiveIdentifying novel biomarkers involved in the development of gastric cancer (GC) can provide potential therapeutic strategies and improve clinical prognosis. miR-301-3p and Cx43 are reportedly dysregulated in GC. miR-301-3p and Cx43 interaction, and their functions in GC progression, are still poorly understood.MethodsThe expression levels of miR-301-3p and Cx43 in GC tissues and cell lines with various differentiation degrees were evaluated by RT-qPCR. The interaction between miR-301-3p and Cx43 was assessed by dual-luciferase reporter assays. CCK8 and Transwell assays were employed to assess the effects of the miR-301-3p-Cx43 axis on GC cell proliferation, migration, and invasion.ResultsCx43 was significantly downregulated in GC tissues and cell lines, while miR-301-3p expression was negatively correlated with Cx43 mRNA levels. The expression levels of Cx43 and miR-301-3p were closely associated with the differentiation, TNM stage, vascular invasion, and lymph node metastasis status of GC patients. Cx43 overexpression could suppress the proliferation, migration, and invasion of GC cells. Cx43 mRNA is a direct target of miR-301-3p, and transfection of an miR-301-3p mimic could reverse the inhibitory effects of Cx43.ConclusionThe miR-301-3p-Cx43 axis is involved in the development and progression of GC by affecting the proliferation, migration, and invasion of GC cells.     

Hsa-miR-145对人三阴性乳腺癌细胞侵袭和迁移的影响

目的 探讨Hsa-miR-145对人三阴性乳腺癌细胞增殖、侵袭及迁移的影响,并初步分析其影响三阴性乳腺癌细胞增殖、侵袭及迁移的可能机制.方法 运用脂质体介导的转染方法将miR-145阻遏物(miR-145 inhibitors)转染人三阴性乳腺癌细胞株MDA-MB-231,以inhibitor negative control(inhibitor NC)作为阴性对照,通过MTT法和Transwell侵袭实验检测细胞的增殖能力和侵袭力;采用Transwell迁移实验及划痕试验检测细胞的迁移能力;利用生物信息学方法预测miR-145的靶基因,并对其靶基因进行基因功能分析.结果 (1)miR-145 inhibitors组细胞增殖活性明显高于inhibitor NC组(P<0.05);(2)划痕后miR-145 inhibitors组细胞迁移能力比inhibitor NC组明显增强(P<0.05);(3)Transwell侵袭及迁移实验均显示转染miR-145 inhibitors后,MDA-MB-231细胞的侵袭及迁移能力明显增强(P<0.01,P<0.05);(4)生物信息学方法预测miR-145的靶基因中,部分发挥了促进细胞增殖、侵袭及迁移的生物学功能.结论 (1)miR-145对人三阴性乳腺癌细胞的增殖、侵袭及迁移能力可能存在负性调控作用;(2)miR-145可能通过多种靶基因发挥其对肿瘤的调控作用.     

DICER-AS1 functions as competing endogenous RNA that targets CSR1 by sponging microRNA-650 and suppresses gastric cancer progression

ObjectiveThis study explored the functional interactions between the long non-coding RNA DICER-AS1 and the cellular stress response 1 (CSR1) gene in gastric cancer.MethodsQuantitative polymerase chain reaction (qPCR) and western blotting were used to measure DICER-AS1, CSR1, and miR-650 expression levels. Gastric cancer cell line proliferation and migration abilities were analyzed using the MTT and transwell migration and invasion assays, respectively. Bioinformatic analysis and dual luciferase reporter assays were employed to study the functional interactions among miR-650, DICER-AS1, and CSR1.ResultsDICER-AS1 and CSR1 expression levels were significantly decreased in gastric cancer tissues compared with normal tissues, and qPCR analysis showed that miR-650 was upregulated in gastric cancer tissues. Bioinformatic analysis and dual luciferase reporter assays revealed that DICER-AS1 functioned as a competing endogenous RNA that sponged miR-650, which in turn regulated CSR1 expression. Importantly, ectopic DICER-AS1 and CSR1 expression inhibited cell proliferation and migration in vitro and suppressed xenograft tumorgenicity in vivo.ConclusionsThese results suggest that DICER-AS1 functions as a competing endogenous RNA that regulates miR-650 to suppress proliferation and migration of gastric cancer cells by targeting CSR1. These findings indicate that targeting DICER-AS1 and miR-650 could be a novel treatment for gastric cancer.     

The role of microRNA-572 in the proliferation and chemotherapeutic treatment of prostate cancer

ObjectiveMicroRNAs (miRNAs) regulate prostate tumorigenesis and progression by involving different molecular pathways. In this study, we examined the role of miR-572 in prostate cancer (PCa).MethodsThe proliferation rates of LNCaP and PC-3 PCa cells were studied using MTT assays. Transwell migration and Matrigel invasion assays were performed to evaluate cell migration and invasion, respectively. Protein expression levels were examined using western blotting. Docetaxel-induced apoptosis was evaluated by Caspase-Glo3/7 assays. The putative miR-572 binding site in the phosphatase and tensin homolog (PTEN) 3ʹ untranslated region (3ʹ UTR) was assessed with dual-luciferase reporter assays. Additionally, miR-572 expression levels in human PCa tissues were examined by qRT-PCR assays.ResultsUpregulation of miR-572 promoted proliferation, migration, and invasion of PCa cells. Overexpression of miR-572 decreased sensitivity of PCa cells to docetaxel treatment by reducing docetaxel-induced apoptosis. MiR-572 can regulate migration and invasion in PCa cells. Furthermore, miR-572 could regulate expression of PTEN and p-AKT in PCa cells by directly binding to the PTEN 3ʹ UTR. MiR-572 expression levels were increased in human PCa tissues and associated with PCa stage.ConclusionsmiR-572 displayed essential roles in PCa tumor growth and its expression level may be used to predict docetaxel treatment in these tumors.     

肿瘤抑素对婴幼儿血管瘤内皮细胞增殖活力与细胞周期的影响

目的探讨抗血管生成药物肿瘤抑素(tumstatin)对体外培养的婴幼儿血管瘤内皮细胞增殖与细胞周期的影响。方法将切取的血管瘤组织在体外消化,获得血管瘤内皮细胞,进行体外培养和传代;以不同浓度tumstatin多肽加入细胞培养基中,MTT法检测细胞活力,以流式细胞仪检测细胞周期的变化。结果在2.5%体积胎牛血清的培养条件下,tumstatin对血管瘤内皮细胞增殖活力的抑制,呈现出量效关系;在17.0μg/mL浓度tumstatin的作用下,血管瘤内皮细胞的细胞周期G0/G1期为55.8%,而对照组为42.9%。结论 tumstatin能够在体外有效地抑制婴幼儿血管瘤内皮细胞的增殖活力,并呈现出一定的量效关系;tumstatin还能影响细胞周期的进程,促使其阻滞在G0/G1期。