Long non-coding RNA small nucleolar RNA host gene 7 facilitates the proliferation,migration, and invasion of esophageal cancer cells by regulating microRNA-625

BackgroundEsophageal cancer (EC) is a highly aggressive malignant tumor, of which esophageal squamous cell carcinoma (ESCC) constitutes the main subtype. Long non-coding RNA (lncRNA) small nucleolar RNA host gene 7 (SNHG7) has been extensively studied in many tumors and has been confirmed to be an oncogene; however, it has yet to be investigated in an ESCC study. Therefore, this study intended to uncover the role of SNHG7 in ESCC.MethodsQuantitative real-time polymerase chain reaction was applied to measure the expression levels of SNHG7 and miR-625 in ESCC tumor tissues and cell lines. Cell Counting Kit-8 assay, 5-Ethynyl-2''-deoxyuridine assay, scratch assay, and Transwell assay were conducted to assess the proliferation, migration, and invasion ESCC cell. We verified the interaction between SNHG7 and miR-625 by performing the dual luciferase reporter gene experiment.ResultsCompared to that in adjacent normal tissues and HET1A cell lines, the expression level of SNHG7 in ESCC tumor tissues and ESCC cell lines was up-regulated, while the expression level of miR-625 was down-regulated. ESCC cell proliferation, migration, and invasion were significantly promoted by SNHG7 overexpression but inhibited by silencing of SNHG7. Further, luciferase reporter gene experiments confirmed that SNHG7 interacted with miR-625, and rescue experiments showed that SNHG7 promoted the malignant phenotype by inhibiting miR-625.ConclusionsSNHG7 is up-regulated in ESCC tumor tissues and cell lines, while miR-625 is expressed at a low level. SNHG7 is able to facilitate the proliferation, migration, and invasion of ESCC cells by targeting miR-625.     

lncRNA SNHG3 acts as oncogene in ovarian cancer through miR-139-5p and Notch1

Ovarian cancer (OC) is a common malignant tumor of the female reproductive system. Long non-coding RNAs (lncRNAs) play an important role in OC occurrence and development. Thus, the function and potential mechanism of lncRNA small nucleolar RNA host gene 3 (SNHG3) was explored in the development of OC. The expression of SNHG3, microRNA (miR)-139-5p and Notch homolog 1, translocation-associated (Drosophila) (Notch1) in OC were detected by RT-qPCR or western blot assay. In addition, CCK-8 and wound-healing assays were used to detect OVCAR3 proliferation and migration ability. The targeting relationship of miR-139-5p with SNHG3 or Notch1 was verified through luciferase reporter assay. Rescue experiments were performed to confirm whether SNHG3 could mediate OVCAR3 proliferation and migration through miR-139-5p and Notch1. In OC tissues and cell lines, the expression of SNHG3 and Notch1 were significantly increased, and the expression of miR-139-5p was significantly decreased. SNHG3 inhibition suppressed the proliferation and migration of OVCAR3 cells. Luciferase reporter experiment confirmed that miR-139-5p could target SNHG3 and Notch1. Transfection of miR-139-5p inhibitor significantly reversed the inhibitory effect of SNHG3 knockdown on OVCAR3 proliferation and migration. Moreover, SNHG3 inhibition or miR-139-5p mimic abolished the promotion of Notch1 overexpression on OVCAR3 proliferation and migration. In conclusion, SNHG3 could accelerate the proliferation and migration of OC cells by regulating miR-139-5p and Notch1.     

miR-138-5p靶向调控RAB22A表达抑制胶质瘤细胞的增殖、迁移和侵袭

目的:探讨miR-138-5p对胶质瘤细胞增殖、迁移与侵袭的影响以及其作用机制。方法:U251细胞分为miR-NC组(对照组)、miR-mimics组、miR-inhibitor组以及miR-mimics+RAB22A-pcDNA3.1组。qRT-PCR检测miR-138-5p在胶质瘤细胞中的表达量;CCK8实验检测U251细胞的增殖能力;Transwell实验检测U251细胞的迁移和侵袭能力;Targetscan数据库预测miR-138-5p的下游靶基因;运用双荧光素酶报告基因实验验证细胞周期蛋白RAB22A是miR-138-5p的靶基因;RNA结合蛋白免疫沉淀实验进一步验证miR-138-5p与RAB22A之间的相互作用;运用蛋白免疫印迹实验检测组织和细胞中RAB22A的蛋白表达水平。结果:与正常人星形胶质细胞相比较,miR-138-5p 在胶质瘤细胞中表达量降低(P<0.05);CCK8和Transwell实验结果表明,在U251细胞中,过表达miR-138-5p能显著抑制细胞的增殖、迁移和侵袭能力(P＜0.05);运用Targetscan数据库预测miR-138-5p下游靶基因为RAB22A,miR-138-5p作用于RAB22A的3' UTR区域,双荧光素酶报告基因实验与蛋白免疫沉淀实验结果证实miR-138-5p与RAB22A之间的相互作用;RAB22A在胶质瘤细胞中明显高表达(P＜0.05),在U251细胞中过表达miR-138-5p明显抑制RAB22A的表达(P＜0.05)。结论:miR-138-5p在胶质瘤细胞中表达降低,miR-138-5p通过下调RAB22A表达而抑制细胞增殖、迁移和侵袭。     

lncRNA LUADT1靶向miR-138-5p调控胶质瘤细胞的增殖、迁移和侵袭

目的:探讨肺腺癌相关转录本1(LUADT1)对胶质瘤细胞的增殖、迁移和侵袭的影响及分子机制.方法:将si-NC、si-LUADT1、miR-NC、miR-138-5p、pcDNA、pcDNA-LUADT1分别转染至U251细胞中,记为si-NC组、si-LUADT1组、miR-NC组、miR-138-5 p组、pcDN...     

长链非编码RNA SNHG1靶向miR-340-5p/CCND1轴调控食管癌细胞增殖、迁移和侵袭

目的:分析长链非编码RNA小核仁RNA宿主基因1（LncRNA SNHG1）靶向miR-340-5p/细胞周期蛋白1（CCND1）轴调控食管癌细胞增殖、迁移和侵袭。方法:体外培养人食管上皮细胞、食管癌细胞KYSE-30、TE-1、NEC、Eca109,qRT-PCR法测定细胞中LncRNA SNHG1、miR-340-5p、CCND1 mRNA水平。将对数期NEC细胞分为对照组、sh NC1组、sh LncRNA SNHG1组、sh NC2组、sh CCND1组、sh LncRNA SNHG1+miR-340-5p inhibitor组、sh CCND1+miR-340-5p inhibitor组。CCK-8法测定细胞增殖能力,Transwell小室法测定细胞侵袭、迁移能力,Western blot法检测CCND1、Ki-67、MMP-2蛋白表达,双荧光素酶验证miR-340-5p与LncRNA SNHG1、CCND1的靶向关系,通过裸鼠瘤内注射转染试剂进行体内试验。结果:与人食管上皮细胞相比,食管癌细胞KYSE-30、TE-1、NEC、Eca109中LncRNA SNHG1、CCND1 mRNA表达升高,miR-340-5p表达降低（P＜0.05）,其中NEC细胞变化最显著,所以使用NEC作为下续研究菌株。与对照组、sh NC组相比,sh LncRNA SNHG1组NEC细胞LncRNA SNHG1、OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2降低（P＜0.05）;与对照组、sh NC组相比,sh CCND1组CCND1 mRNA与蛋白表达、OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2表达降低（P＜0.05）。miR-340-5p与LncRNA SNHG1、CCND1均靶向结合,与sh LncRNA SNHG1组相比,sh LncRNA SNHG1+miR-340-5p inhibitor组OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2蛋白表达升高（P＜0.05）;与sh CCND1组相比,sh CCND1+miR-340-5p inhibitor组OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2蛋白表达升高（P＜0.05）;裸鼠移植瘤实验进行了体内验证。结论:LncRNA SNHG1沉默可能通过调控miR-340-5p/CCND1表达抑制食管癌NEC细胞增殖、侵袭与迁移,裸鼠体内也验证了这一结果。     

miR-590-5p介导lncRNA SNHG1信号影响卵巢癌细胞增殖的实验研究

目的:探讨miR-590-5p在卵巢癌组织中的表达情况以及其对卵巢癌细胞增殖的影响及作用机制。方法:Real-time PCR和双荧光素酶实验确定miR-590-5p与lncRNA SNHG1之间的调控作用。Real-time PCR检测卵巢癌、癌旁组织以及卵巢癌细胞中miR-590-5p的表达。分别采用NC mimic或者miR-590-5p mimic转染两株卵巢癌细胞，CCK-8检测各组细胞的增殖情况。此外，Real-time PCR检测两株细胞中SOX2、RECK和YAP1的表达。结果:Real-time PCR结果显示下调SNHG1后miR-590-5p的表达显著升高。双荧光素酶结果显示转染miR-590-5p mimic和野生型SNHG1片段的细胞中荧光素酶的活性显著降低。此外，Real-time PCR结果显示miR-590-5p在卵巢癌组织中的表达水平显著低于癌旁组织。CaOV3、OV-90细胞中miR-590-5p的表达水平明显低于其他卵巢癌细胞。转染miR-590-5p mimic显著上调了CaOV3、OV-90细胞中miR-590-5p的表达并且抑制了这两株细胞的增殖，同时抑制了两株细胞中SOX2、RECK和YAP1的表达。结论:miR-590-5p的低表达与卵巢癌的进展密切相关，miR-590-5p能够介导SNHG1信号并且通过对其下游靶基因的调控抑制卵巢癌细胞的增殖。     

miR-143-3p通过靶向EZH2 调控结肠癌RKO细胞增殖、迁移和侵袭

目的：探讨miR-143-3p 通过靶向果蝇zeste 基因增强子同源物2（enhancer of zeste homolog 2, EZH2）调控结肠癌RKO细胞增殖、迁移和侵袭的分子机制。方法：选用2015 年3 月至2017 年7 月昆明医科大学第一附属医院手术切除的40 例结肠癌患者的癌及癌旁组织标本,以及结肠癌细胞系COLO320、RKO、CL-11 和正常肠黏膜细胞株NCM460,用qPCR 法检测结肠癌组织和细胞系中miR-143-3p 的表达水平。分别将miR-143-3p mimics、miR-143-3p inhibitor、EZH2 shRNA 及阴性对照质粒转染进RKO细胞,用CCK-8 法、Transwell 小室法分别检测miR-143-3p/EZH2 分子轴对RKO细胞增殖、迁移和侵袭的影响,用Western blotting 检测RKO细胞中EZH2蛋白的表达。用双荧光素酶报告基因实验验证miR-143-3p 和EZH2 的靶向关系。结果：miR-143-3p在结肠癌组织和细胞系中均低表达（均P<0.01）。过表达miR-143-3p 显著抑制RKO 细胞的增殖、迁移和侵袭能力（均P<0.01）。双荧光素酶报告基因实验证实miR-143-3p 靶向EZH2。同时敲降miR-143-3p 和EZH2 可逆转敲降EZH2 对RKO细胞增殖、迁移和侵袭能力的抑制作用。结论：miR-143-3p 通过靶向EZH2并下调其表达水平进而抑制结肠癌细胞的增殖、迁移与侵袭。     

lncRNA XIST 通过miR-32-5p/EZH2 分子轴调控结直肠癌HCT-8细胞的恶性生物学行为

目的：探讨lncRNA XIST 通过miR-32-5p/果蝇Zeste 基因增强子同源物2（enhancer of zeste homolog 2,EZH2）分子轴调控结直肠癌HCT-8 细胞的恶性生物学行为。方法：收集2014 年7 月至2018 年8 月中南大学湘雅医院直肠肛门外科资料完整的结直肠癌患者28 例癌组织和配对的癌旁组织标本,采用qPCR检测结直肠癌组织及细胞系中lncRNA XIST 和miR-32-5p 的表达水平,双荧光素酶报告基因验证lncRNA XIST、miR-32-5p 和EZH2 的靶向关系,并进一步通过WB检测EZH2 的表达水平。CCK-8、Transwell 及Annexin V-FITC/PI 染色流式细胞术检测HCT-8 细胞增殖、迁移及凋亡情况。结果：lncRNA XIST 在结直肠癌组织及细胞系中高表达,且在HCT-8 细胞中表达最高（P<0.05 或P<0.01）。双荧光素酶报告基因证实,lncRNA XIST 靶向负调控miR-32-5p（P<0.05）,且EZH2 是miR-32-5p 的靶基因。敲降lncRNA XIST 抑制HCT-8 细胞增殖和迁移并诱导其凋亡（P<0.05或P<0.01）;进一步实验证明,敲降lncRNA XIST 上调miR-32-5p 的表达水平,从而下调EZH2 表达水平,进而抑制HCT-8 细胞增殖和迁移并诱导其凋亡。结论：lncRNA XIST通过miR-32-5p/EZH2 分子轴促进HCT-8 细胞增殖、迁移并抑制细胞凋亡。     

LncRNA SNHG5 affects cell proliferation,metastasis and migration of colorectal cancer through regulating miR-132-3p/CREB5

We aimed at the effects of long non-coding RNA (lncRNA) SNHG5 on proliferation, metastasis and migration of colorectal cancer (CRC) cells. We also investigated regulatory relationships among miR-132-3p, SNHG5 and CREB5 and their roles in CRC. 25 pairs of samples containing CRC tissues and matched para-tumor tissues were obtained to examine SNHG5, miR-132-3p and CREB5 expression by qRT-PCR or Western blot. The targeted relationship between miR-132-3p and SNHG5 or CREB5 was confirmed by dual luciferase report assay as well as RNA pull down assay. The expression of SNHG5, miR-132-3p and CREB5 in CRC cells were regulated by cell transfection. CRC cellular proliferation was assayed by CCK-8 and meanwhile flow cytometry was adopted to observe apoptosis. Metastasis and migration of CRC cells were determined respectively by means of Transwell assay and scratch test. The effects of SNHG5 on CRC were researched in vivo, too. SNHG5 or CREB5 was up-regulated in CRC tissues and cells, whereas miR-132-3p was down-regulated. Overexpression of SNHG5 and CREB5 resulted in the enhancement of proliferation, metastasis, migration and the inhibition of apoptosis in CRC cells, while miR-132-3p led to the opposite result. LncRNA SNHG5 promoted proliferation, migration and metastasis of CRC cells but inhibited apoptosis by modulating miR-132-3p/CERB5.     

Downregulation of circ-TRPS1 suppressed prostatic cancer prognoses by regulating miR-124-3p/EZH2 axis-mediated stemness

Abnormal circular RNA (circRNA) expression correlates with human traits such as many kinds of cancers. Though circRNAs have links to cancer, they have less characterization in metastatic castration-resistant prostate cancer (PCa), which is main reason for PCa mortality. Therefore, high-throughput sequencing was used for selected circRNA profiles. The result showed that circ-TRPS1 was upregulated significantly in high-grade PCa tissues or cell lines. High circ-TRPS1 expression correlated to aggressive PCa phenotypes. Knockdown of circ-TRPS1 suppressed PCa proliferation and metastasis through targeting miR-124-3p/EZH2 axis-mediated stemness in PCa, which was validated by luciferase reporter assays. EZH2 overexpression or miR-124-3p inhibition reversed the inhibition of circ-TRPS1 silencing in PCa cell migration and proliferation by recovering stemness. In summary, data demonstrated that circ-TRPS1 suppressed PCa progression through functioning similar to a miR-124-3p sponge to enhance EZH2 expression and cancer stem-like cell differentiation. Thus, circ-TRPS1 might be a candidate target for PCa treatment.     

MiR-138 induces cell cycle arrest by targeting cyclin D3 in hepatocellular carcinoma

The deregulation of microRNA (miRNA) is frequently associated with a variety of cancers, including hepatocellular carcinoma (HCC). In this study, we identified 10 upregulated miRNAs (miR-217, miR-518b, miR-517c, miR-520g, miR-519a, miR-522, miR-518e, miR-525-3p, miR-512-3p and miR-518a-3p) and 10 downregulated miRNAs (miR-138, miR-214, miR-214#, miR-27a#, miR-199a-5p, miR-433, miR-511, miR-592, miR-483-5p and miR-483-3p) by Taqman miRNAs array and quantitative real-time PCR (qRT-PCR) confirmation. Additionally, we investigated the expression and possible role of miR-138 in HCC. qRT-PCR results showed that miR-138 was downregulated in 77.8%(14/18) of HCC tissues compared with adjacent non-tumor tissues. Overexpression of miR-138 reduced cell viability and colony formation by induction of cell arrest in HCC cell lines and inhibited tumor cell growth in xenograft nude mice. The use of miR-138 inhibitor increased cell viability and colony formation in HCC cell lines and tumor cell growth in xenograft nude mice. Using TargetScan predictions, CCND3 was defined as a potential direct target of miR-138. Furthermore, CCND3 protein expression was observed to be negatively correlated with miR-138 expression in HCC tissues. The dual-luciferase reporter gene assay results showed that CCND3 was a direct target of miR-138. The use of miR-138 mimic or inhibitor could decrease or increase CCND3 protein levels in HCC cell lines. We conclude that the frequently downregulated miR-138 can regulate CCND3 and function as a tumor suppressor in HCC. Therefore, miR-138 may serve as a useful therapeutic agent for miRNA-based HCC therapy.     

lncRNA SNHG11通过吸附miR-193a-5p促进NSCLC细胞A549的恶性生物学行为

目的：探讨lncRNA SNHG11对非小细胞肺癌（NSCLC）A549细胞增殖、侵袭和迁移的影响及其可能机制。方法:qPCR检测人胚肺细胞HEL-1和NSCLC细胞A549、H1299、HCC827中lncRNA SNHG11和miR-193a-5p的表达水平,向A549细胞中转染SNHG11小干扰RNA(si-SNHG11)、miR-193a模拟物（miR-193a mimic）或miR-193a抑制剂（miR-193a inhibitor）后,CCK-8法检测其对细胞增殖的影响,Transwell小室和细胞划痕实验检测对细胞侵袭和迁移的影响,WB法检测对细胞增殖抗原Ki67、细胞周期蛋白D1(cyclin D1)表达的影响,双荧光素酶报告实验验证lncRNA SNHG11与miR-193a-5p的靶向关系。结果：与人胚肺细胞HEL-1相比,NSCLC细胞A549、H1299、HCC827中lncRNA SNHG11均呈高表达、miR-193a-5p呈低表达（均P<0.05）;沉默lncRNA SNHG11可抑制A549细胞的增殖、侵袭和迁移,降低细胞中Ki67和Cyclin...     

LncRNA WEE2-AS1 knockdown inhibits the proliferation,migration and invasion of glioma cells via regulating miR-29b-2- 5p/TPM3 axis

Glioma is a general malignant tumor with a dismal prognosis. Long noncoding RNAs (lncRNAs) have been implicated in the initiation and processes of tumors. An investigation of the GEPIA database revealed that long noncoding RNA WEE2 antisense RNA 1 (WEE2-AS1) is upregulated in glioma tissues compared to normal brain tissues, and validation with quantitative real-time polymerase chain reaction (qRT–PCR) revealed that WEE2-AS1 expression was consistent with the database prediction. Fluorescence in situ hybridization (FISH) assays revealed that WEE2-AS1 was localized primarily in the cytoplasm. Clone formation experiment and EDU assay were used to detect cell proliferation ability, and Transwell assay was used to detect cell migration and invasion ability, Western-blot assay and immunofluorescence were used to determine TPM3 protein level. Functional experiments revealed that the downregulation of WEE2-AS1 impeded cell proliferation, migration, and invasion in glioma cell lines. Furthermore, downregulation of WEE2-AS1 suppressed tumor growth in vivo. Bioinformatics predictions and integrated experiments indicated that WEE2-AS1 promoted tropomyosin 3 (TPM3) expression by sponging miR-29b-2-5p. A dual-luciferase reporter assay was conducted to uncover the binding of WEE2-AS1 and miR-29b-2-5p and that of miR-29b-2-5p and TPM3. Additionally, a series of rescue assays showed that WEE2-AS1 promotes proliferation, migration, and invasion by targeting miR-29b-2-5p to regulate TPM3 expression. Ultimately, the results of this study indicate that WEE2-AS1 plays an oncogenic role in glioma and may promote further investigations of the diagnostic and prognostic value of WEE2-AS1 in glioma.     

miR-340 suppresses glioblastoma multiforme

Deregulation of microRNAs (miRs) contributes to tumorigenesis. Down-regulation of miR-340 is observed in multiple types of cancers. However, the biological function of miR-340 in glioblastoma multiforme (GBM) remains largely unknown. In the present study, we demonstrated that expression of miR-340 was downregulated in both glioma cell lines and tissues. Survival of GBM patients with high levels of miR-340 was significantly extended in comparison to patients expressing low miR-340 levels. Biological functional experiments showed that the restoration of miR-340 dramatically inhibited glioma cell proliferation, induced cell-cycle arrest and apoptosis, suppressed cell motility and promoted autophagy and terminal differentiation. Mechanistic studies disclosed that, miR-340 over-expression suppressed several oncogenes including p-AKT, EZH2, EGFR, BMI1 and XIAP. Furthermore, ROCK1 was validated as a direct functional target miR-340 and silencing of ROCK1 phenocopied the anti-tumor effect of mR-340. Our findings indicate an important role of miR-340 as a glioma killer, and suggest a potential prognosis biomarker and therapeutic target for GBM.