miR-132 Targets FOXA1 and Exerts Tumor-Suppressing Functions in Thyroid Cancer

MicroRNA-132 (miR-132) has been demonstrated to be a tumor suppressor in several types of tumors. However, the expression and the role of miR-132 in human thyroid cancer are still poorly understood. The aim of the present study was to examine the potential roles and molecular mechanism of miR-132 in thyroid cancer. We found that miR-132 expression levels were significantly downregulated in thyroid cancer tissues and cell lines. Function assays showed that overexpression of miR-132 in TPC1 cells inhibited cell proliferation, migration, and invasion. Forkhead box protein A1 (FOXA1) was identified as a direct target of miR-132 in thyroid cancer cells. Knockdown of FOXA1 in TPC1 cells significantly inhibited cell proliferation, migration, and invasion, which mimicked the suppressive effect induced by miR-132 overexpression. Restoration of FOXA1 expression partially reversed the suppressive effect induced by miR-132 overexpression. Taken together, these results suggested that miR-132 acts as a tumor suppressor in thyroid cancer through targeting FOXA1.     

MiR-203 suppresses tumor growth and invasion and down-regulates MiR-21 expression through repressing Ran in esophageal cancer

The expression of miR-203 has been reported to be significantly down-regulated in esophageal cancer. We showed here that overexpression of miR-203 in esophageal cancer cells dramatically increased cell apoptosis and inhibited cell proliferation, migration and invasion as well as tumor growth and down-regulated miR-21 expression. We subsequently identified that small GTPase Ran was a target gene of miR-203. Furthermore, Ran restoration partially counteracted the tumor suppressive effects of miR-203 and increased miR-21 expression. Taken together, our findings suggest that miR-203 may act as novel tumor suppressor in esophageal cancer through down-regulating the expression of Ran and miR-21.     

miR-132 targeting cyclin E1 suppresses cell proliferation in osteosarcoma cells

In this study, we investigated the roles of miR-132 in tumor growth of osteosarcoma. We found that overexpression of miR-132 significantly suppressed in vitro cell proliferation and in vivo tumor growth. In addition, miR-132 overexpression induced G1/S cell cycle arrest of osteosarcoma cells. Further study showed that miR-132 could interact with the 3′-untranslated region of cyclin E1 (CCNE1) gene and repress its expression. Re-expression of CCNE1 (without the 3′UTR) could partially abrogate the miR-132-induced cell proliferation inhibition. Of significance, contrary to CCNE1, expression level of miR-132 was significantly lower in osteosarcoma tissues than in the adjacent normal tissues. Taken together, these results indicate that miR-132 functions as a tumor suppressor in osteosarcoma and that its suppressive effects are mediated chiefly by repressing CCNE1 expression.     

miR-194靶向EZH2抑制甲状腺癌细胞增殖、迁移和侵袭

目的:研究miR-194、EZH2对甲状腺癌细胞增殖、迁移和侵袭的影响,并探讨其作用机制。方法:采用qRT-PCR法检测甲状腺癌组织、癌旁正常组织及甲状腺癌细胞和正常甲状腺上皮细胞中miR-194和EZH2的mRNA表达;将miR-194（转染miR-194 mimics）、miR-NC（未转染细胞）、inhibitor-NC（转染空inhibitor）、miR-194 inhibitor（转染miR-194 inhibitor）、si-EZH2（转染si-EZH2）、miR-194+Vector（miR-194 mimics和pcDNA 3.1共转染）、miR-194+EZH2（miR-194 mimics和pcDNA 3.1-EZH2共转染）均以脂质体法转染到SW579、IHH-4细胞;Western blot检测细胞中EZH2的蛋白表达;MTT法检测细胞的增殖;Transwell检测细胞的迁移和侵袭;双荧光素酶报告基因检测实验检测细胞荧光素酶活性。结果:与正常组相比,甲状腺癌组EZH2的表达显著升高,miR-194的表达显著降低;与人正常甲状腺上皮细胞相比,甲状腺癌细胞中EZH2表达显著升高,miR-194的表达显著降低,且过表达miR-194和沉默EZH2均可抑制甲状腺癌细胞增殖、迁移和侵袭。EZH2为miR-194的靶标,且EZH2可逆转过表达miR-194对甲状腺癌细胞增殖、迁移和侵袭的抑制作用。结论:miR-194可抑制甲状腺癌细胞的增殖、迁移和侵袭,可能与靶向EZH2有关,将可为miR-194靶向治疗甲状腺癌提供依据。     

MicroRNA-212 suppresses tumor growth of human hepatocellular carcinoma by targeting FOXA1

MicroRNA-212 (miR-212) has been reported to play oncogenic or tumor suppressive role in different human malignancies. Here, we demonstrated that the mean level of miR-212 in hepatocellular carcinoma (HCC) tissues was significantly lower than that in matched tumor-adjacent tissues. Similarly, the expression of miR-212 was obviously reduced in HCC cell lines as compared with a nontransformed hepatic cell line. Ectopic expression of miR-212 inhibited cell viability and proliferation, and induced apoptosis in HepG2 cells. In contrast, down-regulation of miR-212 increased cell viability and proliferation, and suppressed apoptosis in Bel-7402 cells. In vivo studies showed that miR-212 inhibited tumor growth of HCC via suppressing proliferation and inducing apoptosis. Furthermore, we confirmed that Forkhead box protein A1 (FOXA1) was a direct target of miR-212, and it abrogated the function of miR-212 in HCC. Finally, we disclosed that the aberrant expression of miR-212 and FOXA1 was evidently correlated with poor prognostic features of HCC. MiR-212, FOXA1 and their combination were valuable prognostic markers for predicting survival of HCC patients. In conclusion, miR-212 may serve as a prognostic indicator for HCC patients and exerts tumor suppressive role, at least in part, by inhibiting FOXA1.     

miR-615 Inhibits Prostate Cancer Cell Proliferation and Invasion by Directly Targeting Cyclin D2

Previous studies have reported that miR-615 exerts a tumor suppressor role in some tumors, such as esophageal squamous cell carcinoma and non-small cell lung cancer. However, the role of miR-615 in prostate cancer has not been defined. Here we found that miR-615 was downregulated in prostate cancer tissues and cell lines. Overexpression of miR-615 in PC-3 cells significantly inhibited cellular proliferation, migration, and invasion. Moreover, overexpression of miR-615 delayed tumor growth in vivo. In terms of mechanism, we found that cyclin D2 (CCND2) is a target gene of miR-615 in prostate cancer. We showed that miR-615 could bind to the 3 -UTR region of CCND2 mRNA and inhibit its expression. There was a negative correlation between the expression of miR-615 and CCND2 in prostate cancer tissues. Moreover, restoration of cyclin D2 abolished the inhibitory effects of miR-615 on the proliferation, migration, and invasion of prostate cancer cells. Taken together, our study identified miR-615 as a tumor suppressor by targeting cyclin D2 in prostate cancer.     

miR-30c Impedes Glioblastoma Cell Proliferation and Migration by Targeting SOX9

miR-30c has been acknowledged as a tumor suppressor in various human cancers, such as ovarian cancer, gastric cancer, and prostate cancer. However, the role of miR-30c in glioblastoma (GBM) needs to be investigated. In our study, we found that the expression of miR-30c was significantly downregulated in GBM tissues and cell lines. We found that overexpression of miR-30c inhibited cellular proliferation of GBM cells in vitro and in vivo. More GBM cells were arrested in the G₀ phase after miR-30c overexpression. Moreover, we showed that miR-30c overexpression suppressed the migration and invasion of GBM cells. Mechanistically, we found that SOX9 was a direct target of miR-30c in GBM cells. Overexpression of miR-30c inhibited the mRNA and protein levels of SOX9 in GBM cells. Moreover, there was a negative correlation between the expression of miR-30c and SOX9 in GBM tissues. Finally, we showed that restoration of SOX9 in GBM cells reversed the proliferation, migration, and invasion of GBM cells transfected with miR-30c mimic. Collectively, our results demonstrated that miR-30c suppressed the proliferation, migration, and invasion of GBM cells via targeting SOX9.     

lncRNA BLACAT1靶向结合miR-29a-3p调控甲状腺癌细胞TPC-1的生物学行为

目的:探讨长链非编码RNA BLACAT1(lncRNA BLACAT1)调控microRNA-29a-3p（miR-29a-3p）对甲状腺癌细胞恶性生物学行为的作用及其可能机制。方法:收集2018年06月至2019年03月在我院行甲状腺癌切除术的31例患者的肿瘤组织和相应的癌旁组织。采用qRT-PCR检测lncRNA BLACAT1、miR-29a-3p在甲状腺癌组织、癌旁组织、5种甲状腺癌细胞系（SW579、 PDTC-1、 HMGA1、 TPC-1、 KAT-5）及正常甲状腺细胞Nthy-ori 3-1中的表达水平;调控TPC-1甲状腺癌细胞系中lncRNA BLACAT1、miR-29a-3p的表达,采用CCK-8法检测细胞增殖情况,Transwell法检测细胞的迁移和侵袭能力。用生物信息分析和双荧光素酶报告基因法预测和验证lncRNA BLACAT1与miR-29a-3p的靶向关系。结果:与癌旁组织和正常甲状腺细胞相比,甲状腺癌组织和细胞系中lncRNA BLACAT1的表达水平显著上调,miR-29a-3p的表达水平显著下调（P＜0.05）;过表达lncRNA BLACAT1促进TPC-1细胞的增殖、迁移和侵袭;在甲状腺癌组织中lncRNA BLACAT1的表达水平与miR-29a-3p的表达水平呈负相关（r2 =0.492,P＜0.001）;双荧光素酶分析证实lncRNA BLACAT1能特异性结合miR-29a-3p,并能降低其表达;过表达miR-29a-3p抑制TPC-1细胞的增殖、迁移和侵袭,且miR-29a-3p可以抑制由lncRNA BLACAT1过表达引起的TPC-1细胞增殖、迁移和侵袭能力的增强。结论:lncRNA BLACAT1通过靶向调控miR-29a-3p表达影响甲状腺癌细胞的增殖、迁移和侵袭,从而促进甲状腺癌的发展。     

LINC00243靶向miR-1976促进甲状腺癌细胞增殖、迁移和侵袭

目的:探讨长链非编码RNA（lncRNA）LINC00243对甲状腺癌细胞增殖、迁移及侵袭的影响和分子机制。方法:实时荧光定量PCR（qRT-PCR）检测正常甲状腺细胞系（HT-ori3）和甲状腺癌细胞系（BCPAP、TPC-1和SW1736）中LINC00243和miR-1976的表达水平。将LINC00243小干扰RNA（si-LINC00243）、miR-1976模拟物（miR-1976 mimics）分别转染TPC-1细胞。细胞计数试剂盒（CCK-8）检测细胞活力;Transwell实验检测细胞迁移和侵袭数量;蛋白质印记（Western blot）检测细胞周期素D1（CyclinD1）、基质金属蛋白酶2（MMP2）和MMP9的表达水平。双荧光素酶报告基因实验和qRT-PCR验证LINC00243和miR-1976的靶向调控关系。结果:与HT-ori3细胞比较,3种甲状腺癌细胞中LINC00243的表达水平显著升高,miR-1976的表达水平显著降低。沉默LINC00243或高表达miR-1976均可抑制TPC-1细胞的增殖、迁移和侵袭,抑制CyclinD1、MMP2和MMP9蛋白的表达（P＜0.05）。LINC00243靶向负性调控miR-1976表达。低表达miR-1976可逆转沉默LINC00243对TPC-1细胞增殖、迁移和侵袭的抑制作用（P＜0.05）。结论:在甲状腺癌细胞中,LINC00243呈高表达,miR-1976呈低表达。LINC00243通过靶向调控miR-1976促进甲状腺癌细胞增殖、迁移和侵袭。     

Caveolin-1 mediates tumor cell migration and invasion and its regulation by miR-133a in head and neck squamous cell carcinoma

MicroRNAs (miRNAs) are small non-coding RNAs of approximately 22 nucleotides that can function as oncogenes or tumor suppressors in human cancer. Down-regulation of the miRNA miR-133a in many type of cancers, and a reduction of cell proliferation, migration, and invasion upon over-expression, suggests that miR-133a is a tumor suppressor. In this study, genome-wide gene expression analysis of HNSCC cells that over-express miR-133a showed that caveolin-1 (CAV1), a multifunctional scaffolding protein, is down-regulated, a result that was confirmed by real-time PCR and Western blot analysis. A luciferase reporter assay revealed that miR-133a is directly bound to CAV1 mRNA. Cancer cell migration and invasion were significantly inhibited in HNSCC cells transfected with si-CAV1. Therefore, CAV1 functions as an oncogene in HNSCC. The identification of tumor suppressive miRNAs and their target genes could provide new insights into potential mechanism of HNSCC carcinogenesis.     

miRNA-335基因启动子区甲基化与胃癌细胞不良生物学行为的关系

目的:探讨miR-335基因启动子区异常甲基化状态对胃癌细胞系中miR-335表达水平的影响,以及miR-335基因启动子区甲基化状态对胃癌细胞侵袭,迁移,以及增殖能力的影响。方法:1株永生化胃黏膜上皮细胞系(GES-1)和4株胃癌细胞系(SGC-7901,MKN-45,BGC-823和AGS)。实时荧光定量PCR(qRT-PCR)检测胃癌细胞株miR-335及CRKL的表达水平。甲基化特异性PCR(MSP)方法检测胃癌细胞株miR-335的基因启动子区甲基化状态。应用MTT方法检测恢复miR-335表达对胃癌细胞增殖能力的影响,Transwell侵袭迁移实验及划痕愈合实验分析恢复miR-335表达对胃癌细胞系侵袭及迁移能力的影响。结果:MSP实验结果表明,MKN-45、SGC-7901和BGC-823细胞系均存在基因启动子区异常的高甲基化状态,AGS细胞系基因启动子区亦呈部分高甲基化状态。去甲基药物5-aza-2′-deoxycytidine处理后胃癌细胞miR-335的表达水平可升高2~3倍。Transwell侵袭迁移实验及划痕愈合实验表明miR-335表达水平恢复后SGC-7901细胞的侵袭和迁移能力明显降低。MTT实验结果表明5-aza-2′-deoxycytidine处理后的SGC-7901细胞系与对照组相比,增殖能力显著降低。结论:miR-335启动子区的异常高甲基化状态抑制了miR-335在胃癌细胞系中的表达,恢复miR-335的表达水平可以抑制胃癌细胞的增殖,迁移和侵袭能力。miR-335为胃癌的肿瘤抑制因子。     

MicroRNA-664 Targets Insulin Receptor Substrate 1 to Suppress Cell Proliferation and Invasion in Breast Cancer

A large number of microRNAs (miRNAs) have been previously demonstrated to be dysregulated in breast cancer (BC), and alterations in miRNA expression may affect the initiation and progression of BC. This study showed that miR-664 expression was obviously reduced in BC tissues and cell lines. Resumption of the expression of miR-664 attenuated the proliferation and invasion of BC cells. The molecular mechanisms underlying the inhibitory effects of BC cell proliferation and invasion by miR-664 were also studied. Insulin receptor substrate 1 (IRS1) was identified as a novel and direct target of miR-664. In addition, siRNAmediated silencing of IRS1 expression mimicked the suppressive effects of miR-664 overexpression in BC cells. Rescue experiments demonstrated that recovered IRS1 expression partially antagonized the inhibition of proliferation and invasion of BC cells caused by miR-664 overexpression. Thus, miR-664 may serve as a tumor suppressor in BC by directly targeting IRS1. Moreover, miR-664 downregulation in BC may contribute to the occurrence and development of BC, suggesting that miR-664 may be a novel therapeutic target for patients with BC.     

miR-623在膀胱癌中的表达及靶向Fascin1抑制膀胱癌细胞迁移和侵袭

目的:探讨miR-623在膀胱癌中的表达及通过靶向Fascin1对膀胱癌细胞迁移和侵袭能力的影响。方法:采用实时荧光定量PCR检测正常膀胱上皮组织、膀胱癌组织、正常永生化膀胱上皮细胞系(SV-HUC-1)和膀胱癌细胞系（T24、UMUC3）中miR-623的表达量;采用脂质体瞬时转染miR-623 mimics,划痕实验和Transwell侵袭实验检测miR-623过表达后膀胱癌细胞迁移、侵袭能力的改变;生物信息学预测miR-623的作用靶蛋白。miR-623过表达后Western blot及双荧光素酶报告基因检测其靶点的表达及结合情况;使用Fascin1特异性siRNA观察膀胱癌细胞迁移和侵袭能力的变化,并同时转染miR-623 inhibitor进行恢复实验。结果:miR-623在膀胱癌中的表达水平显著低于在正常膀胱组织中的表达（P＜0.05）,在膀胱癌细胞系（T24、UMUC3）中的表达水平显著低于正常膀胱细胞系(SV-HUC-1)（P＜0.05）;miR-623过表达显著抑制T24和UMUC3细胞的迁移和侵袭能力。生物信息学预测Fascin1为miR-623的靶基因,在T24和UMUC3细胞中过表达miR-623,能够显著降低Fascin1的蛋白水平;荧光素酶报告基因分析结果证实miR-623作用于Fascin1的3'-UTR。下调Fascin1表达能够抑制膀胱癌T24和UMUC3细胞的迁移和侵袭能力,同时抑制miR-623的表达能够提高细胞的迁移和侵袭能力。结论:miR-623在膀胱癌中表达水平降低,是一个抑癌因子;并可能通过靶向Fascin1调节膀胱癌的侵袭和转移能力。     

By downregulating TIAM1 expression,microRNA-329 suppresses gastric cancer invasion and growth

Gastric cancer (GC) is one of the most common malignant tumors worldwide. Emerging evidence has shown that abnormal microRNAs (miRNAs) expression is involved in tumorigenesis. MiR-329 was previously reported to act as a tumor suppressor or oncogene in some types of cancer. However, its function in gastric cancer (GC) is unclear. Here, we found that miR-329 was down-regulated in GC compared with adjacent controls. Enforced expression of miR-329 inhibited proliferation, migration and invasion of gastric cancer cells in vitro. We identified T lymphoma invasion and metastasis 1 (TIAM1) gene as potential target of miR-329. MiR-329 levels inversely correlated with TIAM1 expression in GC. Importantly, TIAM1 rescued the miR-329-mediated inhibition of cell invasion and proliferation. Finally, reintroduction of miR-329 significantly inhibited tumor formation of GC in the xenograft mice. Our findings suggest that miR-329 is a tumor suppressor and potential therapeutic target of GC     

miR-363-3p Inhibits Osteosarcoma Cell Proliferation and Invasion via Targeting SOX4

miR-363-3p has been shown to suppress tumor growth and metastasis in various human cancers. However, the function of miR-363-3p in osteosarcoma (OS) has not been determined. In our study, we found that the expression of miR-363-3p was significantly downregulated in OS tissues compared with adjacent normal tissues. miR-363-3p expression was associated with the poor overall survival rate of OS patients. Moreover, we found that overexpression of miR-363-3p markedly inhibited the proliferation, migration, and invasion of U2OS and MG63 cells. Moreover, we found that SOX4 was a direct target of miR-363-3p in OS cells. Overexpression of miR-363-3p significantly inhibited the expression of SOX4. Expression levels of miR-363-3p and SOX4 were negatively correlated in OS tissues. Finally, we found that restoration of SOX4 attenuated the suppressive effects of miR-363-3p on the proliferation, migration, and invasion of U2OS and MG63 cells. Therefore, our findings demonstrated that miR-363-3p served as a tumor suppressor in OS tissues by targeting SOX4.