Long noncoding RNA POU6F2-AS1 regulates lung cancer aggressiveness through sponging miR-34c-5p to modulate KCNJ4 expression

It has been extensively reported that long noncoding RNAs (lncRNAs) were closely associated with multiple malignancies. The aim of our study was to investigate the effects and mechanism of lncRNA POU6F2-AS1 in lung adenocarcinoma (LADC).The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets provided us the information of LADC clinical samples. High-regulation of POU6F2-AS1 was presented in LADC tissues compared with adjacent normal tissues, which was correlated with poor outcome of LADC patients. Functional experiments in Calu-3 and NCI-H460 cells showed that POU6F2-AS1 significantly promoted LADC cell proliferation, colony formation, invasion and migration. Moreover, through online prediction, luciferase reporter assay and Pearson’s correlation analysis, we found that POU6F2-AS1 may act as a competing endogenous RNA (ceRNA) of miR-34c-5p and facilitated the expression of potassium voltage-gated channel subfamily J member 4 (KCNJ4). The promoting effect of cell aggressiveness induced by POU6F2-AS1 was enhanced by KCNJ4, whilst was abrogated due to the overexpression of miR-34c-5p. Collectively, POU6F2-AS1 might function as a ceRNA through sponging miR-34c-5p to high-regulate KCNJ4 in LADC, which indicates that POU6F2-AS1 might be a promising therapeutic target with significant prognostic value for LADC treatment.     

LncRNA HAND2-AS1 inhibits 5-fluorouracil resistance by modulating miR-20a/PDCD4 axis in colorectal cancer

BackgroundChemoresistance is one of the main obstacles in the therapy of human cancers, including colorectal cancer (CRC). Long non-coding RNA heart and neural crest derivatives expressed 2-antisense RNA 1 (lncRNA HAND2-AS1) has been demonstrated to be associated with CRC. However, the function of HAND2-AS1 in 5-Fluorouracil (5-FU) resistance of CRC remains unclear.MethodsQuantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression of HAND2-AS1, miR-20a and programmed cell death factor 4 (PDCD4) mRNA. 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay was conducted to evaluate IC50 of 5-FU and cell proliferation. Flow cytometry analysis was used to determine cell apoptosis. Transwell assay was carried out to measure cell migration and invasion. Western blot assay was conducted to examine the protein levels of B-cell lymphoma-2 (Bcl-2), BCL2-Associated X (Bax), matrix metalloprotein 2 (MMP2), MMP9 and PDCD4. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull down assay were utilized to verify the combination between miR-20a and HAND2-AS1. Dual-luciferase reporter assay was used to analyze the association between miR-20a and PDCD4. Murine xenograft assay was used to confirm the function of HAND2-AS1 in vivo.ResultsHAND2-AS1 and PDCD4 were downregulated and miR-20a was upregulated in 5-FU-resistant CRC tissues and cells. HAND2-AS1 suppressed 5-FU resistance, cell proliferation, migration and invasion and promoted cell apoptosis in 5-FU-resistant CRC cells. HAND2-AS1 acted as a sponge of miR-20a to regulate PDCD4 expression. Moreover, HAND2-AS1 suppressed cell progression and 5-FU resistance by upregulating PDCD4 via sponging miR-20a in 5-FU-resistant CRC cells. Besides, HAND2-AS1 inhibited tumor growth in vivo.ConclusionHAND2-AS1/miR-20a/PDCD4 axis inhibited cell progression and 5-FU resistance in 5-FU-resistant CRC cells.     

E2F1-mediated MNX1-AS1-miR-218-5p-SEC61A1 feedback loop contributes to the progression of colon adenocarcinoma

The long noncoding RNA MNX1-AS1 has been reported to facilitate the progression of glioblastoma and ovarian cancer. Nevertheless, the biological roles and underlying mechanisms of MNX1-AS1 in colon adenocarcinoma have not been studied until now. In the current study, MNX1-AS1 was upregulated in colon adenocarcinoma. JASPAR prediction tool showed that E2F1 could bind to the promoter region of MNX1-AS1. The chromatin immunoprecipitation assay and luciferase reporter assay were used to verify the interactions between MNX1-AS1 and E2F1. Then functional assays revealed that downregulation of MNX1-AS1 decreased cell proliferation, migration, and invasion in colon adenocarcinoma, but upregulation of E2F1 reversed the effects. Moreover, subcellular fractionation assay manifested that MNX1-AS1 was enriched in the cytoplasm of colon adenocarcinoma cells, thus we speculated whether MNX1-AS1 could function as a competing endogenous RNA (ceRNA) to play roles in colon adenocarcinoma. Bioinformatics analysis and luciferase reporter assay indicated that MNX1-AS1 could sponge microRNA-218-5p (miR-218-5p). Furthermore, we discovered that SEC61A1 was downstream target of miR-218-5p, and MNX1-AS1 acted as a ceRNA to upregulate the expression of SEC61A1 through sponging miR-218-5p. Finally, rescue assays confirmed that MNX1-AS1 facilitated the progression of colon adenocarcinoma through regulating miR-218-5p/SEC61A1 axis. Taken together, we concluded that E2F1-mediated MNX1-AS1-miR-218-5p-SEC61A1 feedback loop contributed to the progression of colon adenocarcinoma.     

Long noncoding RNA DLX6-AS1 promotes liver cancer by increasing the expression of WEE1 via targeting miR-424-5p

Long noncoding RNAs (lncRNAs) played an important role in tumorigenesis and development of hepatocellular carcinoma (HCC). In this study, we first demonstrated that lncRNA DLX6 antisense RNA 1 (DLX6-AS1) was upregulated in cancer tissues and cells lines compared with normal adjacent and cell line. Knock-down DLX6-AS1 by transfection with small interfering RNA (siRNA) suppressed cell proliferation, migration, and invasion of HCC cells. Cell cycle analysis showed that cells transfected with siRNA were arrested in G0/G1 phase. Then, we performed dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay to show that DLX6-AS1 could bind with miR-424-5p. And cotransfection inhibitor of miR-424-5p with siRNA of DLX6-AS1 could abolish the inhibitory effect of siRNA of DLX6-AS1 on cell proliferation, migration, and invasion. Moreover, we further demonstrated that the oncogene WEE1 G2 checkpoint kinase (WEE1) was the target of miR-424-5p and expression levels of WEE1 were positive correlation with that of DLX6-AS1. Taken together, these results suggested that upregulated DLX6-AS1 promoted cell proliferation, migration, and invasion of HCC through increasing expression of WEE1 via targeting miR-424-5p.     

LBX2-AS1 promotes ovarian cancer progression by facilitating E2F2 gene expression via miR-455-5p and miR-491-5p sponging

LBX2-AS1 is a long non-coding RNA that facilitates the development of gastrointestinal cancers and lung cancer, but its participation in ovarian cancer development remained uninvestigated. Clinical data retrieved from TCGA ovarian cancer database and the clinography of 60 ovarian cancer patients who received anti-cancer treatment in our facility were analysed. The overall cell growth, colony formation, migration, invasion, apoptosis and tumour formation on nude mice of ovarian cancer cells were evaluated before and after lentiviral-based LBX2-AS1 knockdown. ENCORI platform was used to explore LBX2-AS1-interacting microRNAs and target genes of the candidate microRNAs. Luciferase reporter gene assay and RNA pulldown assay were used to verify the putative miRNA-RNA interactions. Ovarian cancer tissue specimens showed significant higher LBX2-AS1 expression levels that non-cancerous counterparts. High expression level of LBX2-AS1 was significantly associated with reduced overall survival of patients. LBX2-AS1 knockdown significantly down-regulated the cell growth, colony formation, migration, invasion and tumour formation capacity of ovarian cancer cells and increased their apoptosis in vitro. LBX2-AS1 interacts with and thus inhibits the function of miR-455-5p and miR-491-5p, both of which restrained the expression of E2F2 gene in ovarian cancer cells via mRNA targeting. Transfection of miRNA inhibitors of these two miRNAs or forced expression of E2F2 counteracted the effect of LBX2-AS1 knockdown on ovarian cancer cells. LBX2-AS1 was a novel cancer-promoting lncRNA in ovarian cancer. This lncRNA increased the cell growth, survival, migration, invasion and tumour formation of ovarian cancer cells by inhibiting miR-455-5p and miR-491-5p, thus liberating the expression of E2F2 cancer-promoting gene.     

Long noncoding MAGI2-AS3 promotes colorectal cancer progression through regulating miR-3163/TMEM106B axis

Colorectal cancer (CRC), is mostly derived from normal colon epithelial cells, and has been reported to be one of most common gastrointestinal malignancies globally. An increasing number of researchers have claimed that long noncoding RNAs (lncRNAs) exert significant functions in tumor progression. Nevertheless, the function of MAGI2-AS3 remains uncertain in CRC. The expression of MAGI2-AS3, miR-3163, and transmembrane protein 106B (TMEM106B) messenger RNA was examined by quantitative real-time polymerase chain reaction. Cell apoptosis was measured by caspase-3 activity test. Cell proliferation was tested by cell-counting kit 8 and 5-ethynyl-2′-deoxyuridine assays. Cell migration was detected by transwell assay. Western blot analysis examined the protein expression of TMEM106B. The expression of Ki-67 was evaluated by immunohistochemistry assay. The binding capacity between miR-3163 and MAGI2-AS3 (or TMEM106B) was studied by radioimmunoprecipitation and luciferase reporter assays. The expression of MAGI2-AS3 and TMEM106B was conspicuously upregulated whereas miR-3163 presented lower expression in CRC cells. MAGI2-AS3 deficiency facilitated cell apoptosis but hampered cell proliferation and migration. MAGI2-AS3 combined with miR-3163 and negatively regulated miR-3163 expression. In addition, the administration of sh-MAGI2-AS3 or miR-3163 mimics suppressed CRC cell growth in vivo. Subsequently, miR-3163 targeted TMEM106B and the transfection of sh-MAGI2-AS3 or miR-3163 mimics downregulated TMEM106B expression. Rescue assays verified that TMEM106B overexpression recovered the effects of MAGI2-AS3 inhibition on cell apoptosis, proliferation, and migration in CRC. MAGI2-AS3 drives CRC progression through regulating miR-3163/TMEM106B axis. This supplies innovative insights on the investigation of molecular mechanism in CRC progression.     

HOXC13-AS promotes breast cancer cell growth through regulating miR-497-5p/PTEN axis

Dysregulated long noncoding RNAs (lncRNAs) remains to be explored in tumorigenesis. LncRNA HOXC13 antisense RNA (HOXC13-AS) has been found as an oncogene in many cancers; however, the role of HOXC13-AS in breast cancer still elusive. In this study, the HOXC13-AS levels and its role in cell proliferation was first measured by real-time quantitative polymerase chain reaction, Cell Counting Kit-8 assay, and colony formation assay. It showed that HOXC13-AS was increased in breast cancer tissues compared with the adjacent normal tissues and upregulated HOXC13-AS promoted the growth of breast cancer cells. Then, we found that the miR-497-5p levels were downregulated in cancer tissues compared with the adjacent tissues and miR-497-5p suppressed breast cancer cell proliferation. Further study showed that HOXC13-AS could function as a “sponge” for miR-497-5p then suppress miR-497-5p expression. Moreover, we next identified that Phosphatase and Tensin homolog (PTEN) is the target of miR-497-5p. Overexpression of miR-497-5p by chemical mimics decreased the expression of PTEN, while downregulation of miR-497-5p by HOXC13-AS rescued the expression of PTEN. Finally, we showed that HOXC13-AS promoted the proliferation of breast cancer cells and tumor growth through miR-497-5p/PTEN axis in vitro and in vivo. Hence, we conclude that HOXC13-AS, which is significantly upregulated in breast cancers, promoted cell proliferation through the suppressed miR-497-5p and further upregulated PTEN.     

LncRNA PDCD4-AS1 alleviates triple negative breast cancer by increasing expression of IQGAP2 via miR-10b-5p

ObjectiveMounting evidence demonstrates that long non-coding RNA (lncRNA) is dysregulated in breast cancers. This study was designed to detect the influences and regulatory mechanism of lncRNA PDCD4-AS1 in triple-negative breast cancer (TNBC).MethodsqRT-PCR and Western blot were utilized to investigate the expression levels of PDCD4-AS1, miR-10b-5p and IQGAP2 in TNBC tissues and cells. Online software and luciferase reporter gene system were employed to testify the interactions among these molecules. Loss and gain of function of PDCD4-AS1, miR-10b-5p or IQGAP2 were performed before MTT and colony formation assay, TUNEL staining in addition to Transwell and scratch assays were applied to measure the cell biological functions.ResultsIn this work, PDCD4-AS1 and IQGAP2 were lowly expressed while miR-10b-5p was strongly expressed in TNBC tissues and cells. PDCD4-AS1 or IQGAP2 overexpression effectively attenuated TNBC cell proliferation, migration and invasion, and increased the apoptosis rate, while this effect was abandoned in response to miR-10b-5p mimics transfection. miR-10b-5p bound to IQGAP2 and acted as a downstream target of PDCD4-AS1.ConclusionOur findings identified lncRNA PDCD4-AS1 as a tumor suppressor in TNBC by regulating IQGAP2 expression via miR-10b-5p, giving a novel insight into the regulatory mechanism of PDCD4-AS1 in the pathogenesis of TNBC.     

Long noncoding RNA MNX1-AS1 contributes to lung cancer progression through the miR-527/BRF2 pathway

Lung cancer belongs to a leading popular and malignant cancer around the world. However, the root mechanism underlying lung cancer progression remains unclear. Recently, long noncoding RNA (lncRNA) has been identified as important for tumorigenesis. LncRNA MNX1-AS1 is proven to regulate colon adenocarcinoma, cervical cancer, glioblastoma, and ovarian cancer. Whether MNX1-AS1 participates in lung cancer needs investigation. In our research, we found that MNX1-AS1 was dramatically upregulated in lung cancer. MNX1-AS1 upregulation indicated poor prognosis in lung cancer patients. Functionally, MNX1-AS1 promoted lung cancer progression through regulating proliferation, migration, and invasion. Mechanistically, MNX1-AS1 was found to locate in the cytoplasm and interact with miR-527. Through inhibiting miR-527 availability, MNX1-AS1 facilitated BRF2 expression. Restoration of BRF2 rescued defects of proliferation, migration, and invasion caused by MNX1-AS1 knockdown. Taken together, our study found a novel signaling pathway, namely MNX1-AS1/miR-527/BRF2 axis, involved in lung cancer progression.     

LncRNA ZNF667-AS1靶向miR-31-5p对食管癌细胞增殖、迁移和凋亡的机制研究

目的 探究长链非编码RNA(lncRNA)ZNF667-AS1通过靶向miR-31-5p对食管癌细胞增殖和迁移的影响及潜在的机制.方法 采用实时荧光定量PCR(qPCR)技术检测ZNF667-AS1在食管癌细胞Eca109、EC1、TE1和正常食管上皮细胞Het-1A的表达水平,并选择表达差异最大的细胞株进行后续实验....     

lncRNA RPL34-AS1通过靶向调控miR-575表达对卵巢癌SKOV3细胞增殖和凋亡的影响

目的探究RPL34-AS1对卵巢癌细胞增殖、迁移的影响及其作用机制。 方法取对数生长期SKOV3细胞用无血清培养基同步化12 h,将pcDNA、pcDNA-RPL34-AS1、si-NC、si-RPL34-AS1、anti-miR-NC、anti-miR-575转染至SKOV3细胞中,分别记为pcDNA组、pcDNA-RPL34-AS1组、si-NC组、si-RPL34-AS1组、anti-miR-NC组、anti-miR-575组;将pcDNA-RPL34-AS1与miR-NC、miR-575分别共转染至SKOV3细胞中,记为pcDNA-RPL34-AS1+miR-NC组、pcDNA-RPL34-AS1+miR-575组。实时荧光定量PCR （RT-qPCR）检测临床组织标本及转染后各组细胞中RPL34-AS1和miR-575的表达水平;双荧光素酶报告实验检测RPL34-AS1和miR-575的靶向关系;四甲基偶氮唑盐比色法（MTT）检测细胞存活率;流式细胞术检测细胞凋亡;蛋白质印迹（Western blot）法检测细胞周期蛋白D1（Cyclin D1）、细胞周期蛋白依赖性激酶抑制剂1A （p21）、B细胞淋巴瘤/白血病-2 （Bcl-2）、Bcl-2相关X蛋白（Bax）蛋白表达水平。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析,组间两两比较采用LSD-t检验。 结果与癌旁组织相比,卵巢癌组织中RPL34-AS1表达水平降低（1.00±0.08比0.47±0.05）,miR-575表达水平升高（1.01±0.07比3.12±0.28）（P < 0.05）。转染si-RPL34-AS1后,细胞活性升高（48 h：0.68±0.06比0.55±0.05;72 h：0.99±0.08比0.71±0.06）,G1期细胞所占比例降低（13.42±1.38比32.15±2.11）,S期细胞所占比例升高（53.75±5.22比34.69±3.41）,细胞凋亡率降低（4.31±0.42比9.25±0.91）,CyclinD1、Bcl-2表达水平升高（0.92±0.08比0.71±0.07;0.86±0.07比0.61±0.06）,p21、Bax表达水平降低（0.13±0.02比0.29±0.03;0.19±0.02比0.31±0.03） （P均< 0.05）。RPL34-AS1靶向调控miR-575,过表达RPL34-AS1或抑制miR-575后可抑制细胞活性,阻滞细胞周期和促进细胞凋亡。miR-575过表达逆转了RPL34-AS1过表达对卵巢癌SKOV3细胞增殖抑制和凋亡促进的作用。 结论过表达RPL34-AS1可抑制卵巢癌SKOV3细胞增殖,促进细胞凋亡,其机制可能与下调miR-575有关。     

lncRNA XIST-miR137-ATG5调节细胞自噬功能在肠癌细胞5-氟胞嘧啶耐药性中的作用

摘要 目的：本文旨在研究长链非编码RNA XIST-miR137-ATG5的相互作用,同时探讨其调节细胞自噬功能与肠癌细胞5-氟胞嘧啶敏感性的关系。方法：实时聚合酶链反应(real time PCR)检测XIST与miR-137在肠癌细胞中的表达;采用脂质体转染法将si-XIST,miR-137转染入肠癌SW480及HCT116细胞中。采用CCK-8检测瞬时转染si-XIST对肠癌细胞增殖及5-FU敏感性的影响;并利用双荧光素酶报告实验检测miR-137与XIST, miR-137与ATG5相互关系。Western blot方法检测XIST- miR137- ATG5对细胞自噬的影响。结果：与正常结肠细胞FHC比较, XIST在结肠癌细胞系明显高表达,miR-137在结肠癌细胞系明显低表达。与阴性对照组比较,转染si-XIST后,SW480及HCT116细胞增殖能力明显受到抑制,对F-5U的敏感性增强,且抑制自噬蛋白Beclin-1及LC3II/LC3 I的表达。miR-137可与XIST,ATG5 3''UTR结合,抑制XIST和ATG5的表达及功能。在结肠癌SW480细胞中共转染miR-137 inhibitor或过表达ATG5可逆转XIST沉默引起的5-FU耐药,同时可逆转因XIST沉默引起的自噬蛋白表达的抑制。结论：LncRNA XIST或可通过调控mir137-ATG促进结直肠癌细胞SW480自噬从而提高其对5-FU的耐药,针对其这一机制,可为将来针对结肠癌的靶向治疗提供一定的实验基础。     

Long noncoding RNA FLVCR1-AS1 aggravates biological behaviors of glioma cells via targeting miR-4731-5p/E2F2 axis

Long noncoding RNAs (lncRNAs) display essential roles in cancer progression. FLVCR1-AS1 is a rarely investigated lncRNAs involved in various human cancers, such as hepatocellular carcinoma and lung cancer. However, its function in glioma has not been clarified. In our study, we found that FLVCR1-AS1 was highly expressed in glioma tissues and cell lines. And upregulation of FLVCR1-AS1 predicted poor prognosis in patients with glioma. Moreover, FLVCR1-AS1 knockdown inhibited proliferation, migration and invasion of glioma cells. Through bioinformatics analysis, we identified that FLVCR1-AS1 was a sponge for miR-4731-5p to upregulate E2F2 expression. Moreover, rescue assays indicated that FLVCR1-AS1 modulated E2F2 expression to participate in glioma progression. Altogether, our research demonstrates that the FLVCR1-AS1/miR-4731-5p/E2F2 axis is a novel signaling in glioma and may be a potential target for tumor therapy.     

LncRNA CERS6-AS1 promotes proliferation and metastasis through the upregulation of YWHAG and activation of ERK signaling in pancreatic cancer

LncRNAs play essential regulatory roles in pancreatic cancer (PC) tumorigenesis and progression. We aimed to investigate the role of lncRNA CERS6-AS1 in PC. CERS6-AS1 expression was determined in PC tissues and cell lines by PCR analysis. The roles of CERS6-AS1 on proliferation, migration, invasion, and epithelial to mesenchymal transition (EMT) were confirmed via CCK-8 assay, EDU assay, transwell assay, wound healing assay, and western blot assay. Besides, the interaction between CERS6-AS1 and their target genes was verified by luciferase report assays and RIP assays. Animal assays and clinical data analysis were performed to validate the functions in vivo. We found that lncRNA CERS6-AS1 was highly expressed in PC tissues and cells. Additionally, high expression of CERS6-AS1 was obviously associated with poor prognosis. Functional assays demonstrated that CERS6-AS1 downregulation significantly inhibited PC cell growth and migration. Moreover, CERS6-AS1 exerted as a molecular sponge for miR-217-5p (miR-217), and miR-217 was confirmed as a potential target of CERS6-AS1. Subsequently, miR-217 suppressed PC cell proliferation and metastasis by directly targeting YWHAG, which interacted with RAF1 and promoted its phosphorylation, leading to RAF1-mediated ERK signaling activation and translocation of phosphorylated ERK from the cytoplasm to the nucleus. Mechanically, CERS6-AS1 silencing significantly inhibited PC cell proliferation and metastasis via a miR-217/YWHAG/RAF1 signaling axis. CERS6-AS1 exerts as a carcinogen in PC to promote malignant features and behaves as a competitive endogenous RNA for miR-217. We identified CERS6-AS1 as a potential biomarker or therapeutic target to improve PC diagnosis and treatment outcomes.Subject terms: Oncogenes, Tumour biomarkers     

Targeting miR-21 enhances the sensitivity of human colon cancer HT-29 cells to chemoradiotherapy in vitro

5-Fluorouracil (5-FU) is a classic chemotherapeutic drug that has been widely used for colorectal cancer treatment, but colorectal cancer cells are often resistant to primary or acquired 5-FU therapy. Several studies have shown that miR-21 is significantly elevated in colorectal cancer. This suggests that this miRNA might play a role in this resistance. In this study, we investigated this possibility and the possible mechanism underlying this role. We showed that forced expression of miR-21 significantly inhibited apoptosis, enhanced cell proliferation, invasion, and colony formation ability, promoted G1/S cell cycle transition and increased the resistance of tumor cells to 5-FU and X radiation in HT-29 colon cancer cells. Furthermore, knockdown of miR-21 reversed these effects on HT-29 cells and increased the sensitivity of HT-29/5-FU to 5-FU chemotherapy. Finally, we showed that miR-21 targeted the human mutS homolog2 (hMSH2), and indirectly regulated the expression of thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD). These results demonstrate that miR-21 may play an important role in the 5-FU resistance of colon cancer cells.     

Long noncoding RNA Linc00339 promotes triple-negative breast cancer progression through miR-377-3p/HOXC6 signaling pathway

Recently, long noncoding RNAs (lncRNAs) have become the key gene regulators and prognostic biomarkers in various cancers. Through microarray data, Linc00339 was identified as a candidate oncogenic lncRNA. We compared the expression levels of Linc00339 in several breast cancer cell lines and normal mammary gland epithelial cell line. The effects of Linc00339 on tumor progression were examined both in vitro and in vivo. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays were applied to evaluate the functions of Linc00339, miR-377-3p, and HOXC6 on cell proliferation. Flow cytometry analysis was used to detect apoptosis and cell cycle distribution. Overall survival (OS) was analyzed using data from The Cancer Genome Atlas and molecular taxonomy of breast cancer international consortium (METABRIC). Dual luciferase assay and RNA immunoprecipitation were performed to confirm the interaction between Linc003339 and miR-377-3p. Linc00339 was increased in breast cancer cell lines compared with the normal epithelial cell. Through in vitro and in vivo experiments, Linc00339 overexpression promoted triple-negative breast cancer (TNBC) proliferation, inhibited cell cycle arrest, and suppressed apoptosis. Silencing of Linc00339 obtained the opposite effects. Mechanistic investigations demonstrated that Linc00339 could sponge miR-377-3p and regulate its expression. Higher expression of miR-377-3p indicated longer OS in breast cancer patients, especially in TNBC patients. Overexpression of miR-377-3p retarded TNBC cell growth through regulating cell cycle distribution and apoptosis. And miR-377-3p was involved in Linc00339-mediated TNBC proliferation through regulating HOXC6 expression. Knockdown of HOXC6 inhibited TNBC progression. In conclusion, our results illuminated that the novel Linc00339/miR-377-3p/HOXC6 axis played a critical role in TNBC progression and might be a promising therapeutic target for TNBC treatment.     

Down-Regulation of ID2-AS1 Alleviates the Neuronal Injury Induced by 1-Methy1-4-Phenylpyridinium in Human Neuroblastoma Cell Line SH-SY5Y Cells Through Regulating miR-199a-5p/IFNAR1/JAK2/STAT1 Axis

We aimed to illustrate the roles and molecular mechanisms of ID2-AS1 in parkinson’s disease (PD). Methods: qRT-PCR detected the expression of ID2-AS1. CCK-8, LDH release assays the effect of ID2-AS1 knockdown on PD cells. Flow cytometry and Western Blot were used to detect the effect of ID2-AS1 inhibition on PD cell apoptosis. ELISA analysis showed that ID2-AS1 inhibition can reduce the inflammation of PD cells. ROS activity assay showed that inhibiting ID2-AS1 attenuated the oxidative stress induced by 1-methy1-4-phenylpyridinium (MPP+). RNA binding protein immunoprecipitation assay showed that ID2-AS1 is mainly located in the cytoplasm. The luciferase reporter assay is used to verify the interaction. In our study, ID2-AS1 was concentration-dependently and time-dependently up-regulated in MPP+?-treated human neuroblastoma cell line SH-SY5Y. ID2-AS1 knockdown enhanced cell proliferation and decreased cell death in PD cells. Knockdown of ID2-AS1 attenuates MPP+?-induced cytotoxicity in SH-SY5Y cells. ID2-AS1 is a sponge of miR-199a-5p. IFNAR1 is a target of miR-199a-5p. Inhibition of miR-199a-5p and overexpression of IFNAR1 alleviate the inhibitory effect of ID2-AS1 knockdown on MPP+?triggered neuronal injury. Inhibition of miR-199a-5p and overexpression of IFNAR1 alleviate the inhibitory effect of ID2-AS1 knockdown on MPP+?-triggered JAK2/STAT1 activation. Overall, down-regulation of ID2-AS1 alleviated the neuronal injury in PD through regulating miR-199a-5p/IFNAR1/JAK2/STAT1 axis.

     

lncRNA MAGI2-AS3下调miR-31-5p抑制肺腺癌细胞的增殖、迁移、侵袭并促进凋亡

目的研究lncRNA MAGI2-AS3对肺癌A549细胞增殖、迁移、侵袭和凋亡的影响和潜在的分子机制。方法根据转染载体不同将A549细胞分为pcDNA3.1组(转染pcDNA3.1)、pcDNA3.1-MAGI2-AS3组(转染pcDNA3.1-MAGI2-AS3)、anti-miR-NC组(转染anti-miR-NC)、anti-miR-31-5p组(转染anti-miR-31-5p)、pcDNA3.1-MAGI2-AS3+miR-NC组(共转染pcDNA3.1-MAGI2-AS3和miR-NC)、pcDNA3.1-MAGI2-AS3+miR-31-5p组(共转染pcDNA3.1-MAGI2-AS3和miR-31-5p mimics)。实时荧光定量PCR(qRT-PCR)检测miR-31-5p和MAGI2-AS3 RNA的表达,四氮唑蓝(MTT)法测定A549细胞增殖活性,Transwell实验检测细胞迁移和侵袭能力,双荧光素酶报告系统验证MAGI2-AS3与miR-31-5p的调控关系,流式细胞术检测细胞凋亡与周期。两组间比较采用独立样本t检验进行分析;多组间比较采用单因素方差分析,组内多重比较采用SNK-q检验。结果与人正常肺细胞HBE相比,肺癌细胞A549中的MAGI2-AS3表达量(0.48±0.03比1.29±0.06)降低,miR-31-5p表达量(1.01±0.05比0.25±0.02)升高;与pcDNA3.1组比较,pcDNA3.1-MAGI2-AS3组A549细胞活力(0.48±0.04比0.77±0.06)、迁移[(81.33±2.87)个比(124.33±3.09)个]和侵袭[(32.00±2.83)个比(53.00±3.27)个]细胞数、S期细胞所占比例(23.01﹪±1.00﹪比32.95﹪±1.06﹪)均降低,凋亡率(19.95﹪±1.25﹪比7.23﹪±0.51﹪)、G0-G1期细胞所占比例(43.58﹪±2.15﹪比33.56﹪±1.23﹪)均升高;与anti-miR-NC组比较,anti-miR-31-5p组A549细胞活力(0.53±0.04比0.78±0.06)、迁移[(76.00±3.74)个比(108.33±2.87)个]和侵袭[(30.00±1.63)个比(42.33±2.05)个]细胞数、S期细胞所占比例(24.43﹪±1.13﹪比32.91﹪±1.08﹪)降低,凋亡率(18.21﹪±1.24﹪比7.29﹪±0.51﹪)、G0-G1期细胞所占比例(41.56﹪±2.19﹪比33.53﹪±1.27﹪)升高,差异有统计学意义(P均<0.05);双荧光素酶报告系统结果显示,MAGI2-AS3靶向负调控miR-31-5p的表达。与pcDNA3.1-MAGI2-AS3+miR-NC组比较,pcDNA3.1-MAGI2-AS3+miR-31-5p组A549细胞活力(0.68±0.06比0.50±0.04)、迁移[(91.00±1.63)个比(52.67±2.62)个]和侵袭[(62.67±2.49)个比(31.67±4.03个)]细胞数升高,凋亡率(10.59﹪±1.0﹪比21.11﹪±1.14﹪)降低,差异有统计学意义(P均<0.05)。结论lncRNA MAGI2-AS3通过靶向miR-31-5p抑制A549细胞的增殖、迁移和侵袭,促进细胞凋亡。lncRNA MAGI2-AS3是肺癌潜在分子治疗靶点。