MicroRNA-485-5p suppresses cell proliferation and invasion in hepatocellular carcinoma by targeting stanniocalcin 2

Increasing evidences indicate that dys-regulation of MicroRNAs contributes to hepatocellular carcinoma. However, the roles of miR-485-5p in HCC are still largely unexplored. In the present study, our quantitative real-time PCR analysis found that miR-485-5p was significantly down-regulated in 50 pairs of human HCC tissues. Moreover, the reduced expression of miR-485-5p was significantly correlated with larger tumor size and more tumor number in patients with HCC. In vitro studies further showed that overexpression of miR-485-5p mimics could inhibit, while its antisense oligos promote cell proliferation and invasion. Results from the dual-luciferase reporter gene assays and western blot further showed that stanniocalcin 2 was a direct target of miR-485-5p. Therefore, our data suggest a novel role for miR-485-5p in the regulation of HCC progression.     

MiR-449a suppresses cell migration and invasion by targeting PLAGL2 in breast cancer

Background

Breast cancer is one of the most common malignancies worldwide. However, the detailed molecular mechanisms underlying breast cancer metastasis are still incompletely clear. MicroRNAs (miRNAs) play a crucial role in cancer metastasis. In this study, we aimed to analyze the expression and function of miR-449a in breast cancer.

MiR-873-5p inhibits cell migration,invasion and epithelial-mesenchymal transition in colorectal cancer via targeting ZEB1

Recent studies have demonstrated that dysregulation of mircoRNAs (miRNAs) greatly affected biological processes of human cancers, including colorectal cancer. As a member of miRNAs family, miR-873-5p has been proved to be a tumor suppressor in some human cancers. Here, we aim to investigate the effects of miR-873-5p on the migration, invasion and epithelial-mesenchymal transition (EMT) of colorectal cancer cells. The low expression of miR-873-5p in colorectal cancer cells was identified by conducting qRT-PCR analysis. Gain of function assays were designed and conducted to demonstrate the specific function of miR-873-5p overexpression in colorectal cancer progression. Transwell assay and western blot assay were conducted and revealed that miR-873-5p inhibited cell migration, invasion and EMT formation. To find the downstream molecular mechanism of miR-873-5p, mechanism assays were designed and performed to find the downstream target of miR-873-5p. ZEB1 (Zinc finger E-box-binding homeobox 1) was certified to be the target of miR-873-5p through bioinformatics analysis, luciferase activity assay and pull-down assay. Finally, rescue assays were carried out to demonstrate the effects of miR-873-5p-ZEB1 axis on the migration, invasion and EMT process of colorectal cancer cells. In conclusion, we confirmed that miR-873-5p suppressed cell migration, invasion and EMT in colorectal cancer via targeting ZEB1.     

MicroRNA-629-5p promotes osteosarcoma proliferation and migration by targeting caveolin 1

Osteosarcoma is a highly malignant tumor that occurs in the bone. Previous studies have shown that multiple microRNAs (miRNAs) regulate the development of osteosarcoma. This study aimed to explore the role of miR-629-5p and its target gene, caveolin 1 (CAV1), in osteosarcoma development. To analyze the expression of miR-629-5p and CAV1 mRNA in osteosarcoma tissues and cell lines, qRT-PCR analysis was performed. Dual-luciferase reporter experiments were subsequently performed to validate the relationship between CAV1 and miR-629-5p. CCK8 assay was used to measure osteosarcoma cell proliferation, and wound-healing assay was performed to study their migratory phenotype. Our findings revealed that miR-629-5p was overexpressed in osteosarcoma tissues and cells, and thereby enhanced cell proliferation and migration. Further, we validated that miR-629-5p targets CAV1 mRNA directly. CAV1 expression, which was negatively correlated with miR-629-5p expression, was found to be downregulated in osteosarcoma tissue samples. Moreover, our data showed that an increase in CAV1 level led to a decline in osteosarcoma cell proliferation and migration, which could be rescued by miR-629-5p upregulation. Overall, our study confirmed that miR-629-5p promoted osteosarcoma proliferation and migration by directly inhibiting CAV1.     

MiR-337-3p suppresses migration and invasion of breast cancer cells by downregulating ESRP1

Breast cancer (BC) is a common malignant tumor in women, and a considerable number of studies show that aberrant expression of miRNA is correlated with BC development. By analyzing TCGA-BRCA database through bioinformatics method, this study disclosed that miR-337−3p was significantly low in BC tissue and might be a cancer inhibitor in BC. To explore the effect and potential mechanism of miR-337−3p in BC, qRT-PCR was used in this study to indicate that the expression of miR-337−3p was downregulated in BC cells. Then, the effects of miR-337−3p on BC cells were detected by western blot, Cell Counting Kit-8 (CCK-8), wound healing and Transwell assays. After upregulating miR-337−3p expression, the cell viability, migration, invasion and epithelial-mesenchymal transition (EMT) of BC cells were markedly inhibited while cell apoptosis remarkably increased. Besides, it was predicted and identified by bioinformatics analysis and dual-luciferase assay that ESRP1 was a target gene of miR-337−3p. Finally, the progression and EMT of BC cells were promoted after upregulating ESRP1 expression level. However, upregulating miR-337−3p as well as ESRP1 reduced the promotion on the malignant phenotype of BC cells. This result revealed that miR-337−3p could inhibit ESRP1 expression to perform its biological functions. In conclusion, it was illustrated in this study that miR-337−3p is a tumor-inhibitor of BC and plays its regulatory role via its downstream gene ESRP1.     

miR-32-5p靶向ZEB1、ZEB2基因调控乳腺癌增殖和侵袭能力的机制

目的 探讨ZEB1和ZEB2在乳腺癌中的作用及miR-32-5 p对乳腺癌调控的分子机制.方法 通过Western blot和qRT-PCR检测乳腺癌和癌旁组织中ZEB1、ZEB2和miR-32-5 p的表达,筛选出ZEB1和ZEB2高表达的乳腺癌细胞系,采用Western blot和qRT-PCR检测敲减效率.应用克...     

MicroRNA-129-5p suppresses proliferation,migration and invasion of retinoblastoma cells through PI3K/AKT signaling pathway by targeting PAX6

BackgroundRetinoblastoma (RB) is the most common primary intraocular malignancy in children. Accumulating evidences have clarified that microRNAs (miRNAs) modulated signaling molecules by acting as oncogenes or tumor-suppressor genes in RB. Thus, in our study, we aimed to investigate the function of miR-129-5p in RB cells through PI3K/AKT signaling pathway by targeting PAX6. Two RB cell lines, Y79 and WERI-Rb-1, were selected in our study, followed by transfection of miR-129-5p inhibitor or si-PAX6 to explore the regulatory role of miR-129-5p in RB cell proliferation, invasion and migration.Material and methodsDual-luciferase assay was used for the detection of targeting relationship between miR-129-5p and PAX6. Besides, western blot analysis was applied to detect expression of cell cycle-related factors (CDK2 and Cyclin E) and PI3K/AKT signaling pathway-related factors (p-AKT and AKT). Nude mice tumorigenesis experiment was used to evaluate the effect of miR-129a-5p on RB growth in vivo.ResultsmiR-129-5p was down-regulated in RB cell lines. miR-129-5p directly targeted the 3′-untranslated region of PAX6. Artificial down-regulation of miR-129-5p promoted cell proliferation, migration and invasion in RB cell lines Y79 and WERI-Rb-1, and promoted RB growth in vivo via PI3K/AKT signaling pathway, which could be reversed by transfection with silencing PAX6.ConclusionThis study provides evidences that RB progression was suppressed by overexpressed miR-129-5p via direct targeting of PAX6 through PI3K/AKT signaling pathway, which may provide a molecular basis for better treatment for RB.     

miR-34c-3p inhibits cell proliferation,migration and invasion of hepatocellular carcinoma by targeting MARCKS

Recent studies have shown that microRNA-34c-3p (miR-34c-3p) is down-regulated in various types of cancers and involved in tumor growth, invasion and metastasis. However, the roles of miR-34c-3p in hepatocellular carcinoma (HCC) are poorly understood. In this study, the expression profile of miR-34c-3pin HCC tissues and cell lines were examined by quantitative real-time polymerase chain reaction (qRT-PCR). The correlations of miR-34c-3p expression and clinicopathological characteristics were analyzed. The biological role of MiR-34c-3pin cell proliferation, migration and invasion was examined. In addition, the targets of miR-34c-3p were identified. The results showed that miR-34c-3p expression was significantly down-regulated in HCC tissues and cell lines; low expression level of miR-34c-3p was correlated with vascular invasion and advanced TNM stage. In vitro functional assays showed that overexpression of miR-34c-3pin HepG2 and Huh7 cells significantly reduced cell proliferation, migration and invasion. Furthermore, target analysis and luciferase assay identified myristoylated alanine-rich protein kinase c substrate (MARCKS) as a specific target of miR-34c-3p. Knockdown of MARCKS in HepG2 cells reduced cell migration and invasion, but not cell proliferation. Taken together, our findings implicate the potential application of miR-34c-3p as a tumor suppressor in cancer therapy.     

LncRNA NCK1-AS1 promotes non-small cell lung cancer progression via regulating miR-512-5p/p21 axis

ObjectiveThis study aimed at probing into the effect of lncRNA NCK1-AS1 on proliferation, migration and invasion of non-small cell lung cancer (NSCLC) cells and its regulatory function on miR-512-5p/p21 molecular axis.MethodsQuantitative real-time polymerase chain reaction (qRT-PCR) was used to assess the expressions of NCK1-AS1 and miR-512-5p in NSCLC tissues and cell lines. The alterations of cell proliferation, migration, invasion and cell cycle were examined by cell counting kit-8 (CCK-8) assay, BrdU experiment, Transwell experiment and flow cytometry, respectively. The dual-luciferase reporter assay and RNA immunoprecipitation experiment were performed to validate the binding relationships between miR-512-5p and NCK1-AS1, and miR-512-5p the 3'UTR of p21 mRNA. Western blot was used to determine the effects of NCK1-AS1 and miR-512-5p on p21 protein expression.ResultsNCK1-AS1 expression was up-regulated in NSCLC tissues and cells, and its high expression was correlated with shorter overall survival time and faster progression of patients. Overexpression of NCK1-AS1 promoted NSCLC cell proliferation, migration and invasion, and accelerated the cell cycle, whereas NCK1-AS1 siRNA inhibited these malignant biological behaviors, and arrested cell cycle. NCK1-AS1 could bind to miR-512-5p, p21 was verified as a target gene of miR-512-5p, and NCK1-AS1 could up-regulate the expression of p21 in NSCLC cells via repressing miR-512-5p expression.ConclusionNCK1-AS1 promotes NSCLC progression by regulating miR-512-5p/p21 molecular axis.     

MiR-138-5p suppresses lung adenocarcinoma cell epithelial-mesenchymal transition,proliferation and metastasis by targeting ZEB2

Background

MiR-138-5p is regarded as a tumour suppressor in many cancers. Transforming growth factor beta (TGF-β) often acts as a tumor promotor at the late stages of human cancers. However, the function of miR-138-5p on lung adenocarcinoma cells induced by TGF-β remains to be further confirmed.

MiR-137 suppresses migration and invasion by targeting EZH2-STAT3 signaling in human hepatocellular carcinoma

Background

Hepatocellular carcinoma (HCC) is one of the most common and aggressive human malignancies and many cell-intrinsic identities and extrinsic epigenetic factors influence the metastatic potential of HCC cells. MicroRNA-137 is often found to be acting as tumor suppressors, however, how miR-137 involved in the metastasis progression in human HCC remains unclear.

miR-409-3p靶向CXCL9调控滋养层细胞的增殖、迁移和侵袭

目的探讨微小RNA-409-3p(miR-409-3p)对滋养层细胞增殖、迁移和侵袭影响及作用机制。方法实时荧光定量PCR(RT-qPCR)检测正常妊娠胎盘组织和子痫前期胎盘组织中miR-409-3p和干扰素伽玛诱导的单核细胞因子(CXCL9)mRNA表达水平,蛋白印迹(Western blot)法检测CXCL9蛋白表达水平。转染miR-409-3p模拟物或CXCL9小干扰RNA至滋养层细胞HTR8/SVneo,构建miR-409-3p过表达或CXCL9表达抑制的HTR8/SVneo细胞,四甲基噻唑蓝染色法(MTT)检测细胞增殖,Transwell检测细胞迁移和侵袭,Western blot检测细胞周期蛋白D1(CyclinD1)、p21、基质金属蛋白酶2(MMP-2)和MMP-9蛋白表达水平。双荧光素酶报告基因实验验证miR-409-3p与CXCL9调控关系。结果与正常妊娠胎盘组织比较,子痫前期胎盘组织中miR-409-3p水平升高(P<0.05),CXCL9 mRNA和蛋白水平降低(P<0.05)。miR-409-3p过表达或干扰CXCL9表达后,HTR8/SVneo细胞培养48 h和72 h后OD值、迁移和侵袭数、CyclinD1、MMP-2和MMP-9蛋白表达水平降低(P<0.05),p21蛋白表达水平升高(P<0.05)。miR-409-3p在HTR8/SVneo细胞中靶向负调控CXCL9表达。CXCL9过表达逆转了miR-409-3p过表达对HTR8/SVneo细胞增殖、迁移和侵袭的影响。结论 miR-409-3p抑制HTR8/SVneo细胞的增殖、迁移和侵袭与下调CXCL9表达有关。     

miR-128-3p过表达靶向MAPK1抑制膀胱癌5637细胞株的侵袭，迁移和上皮间质转化

目的　研究miR-128-3p过表达对膀胱癌5637细胞株的侵袭,迁移和上皮间质转化影响。 方法　基因预测软件TargetScan筛选出miR-128-3p的靶基因,荧光素酶报告实验验证;RT-PCR检测miR-128-3p和MAPK1的表达,Transwell检测细胞侵袭情况,划痕实验检测细胞迁移能力,Western blot检测E-cadherin、N-cadherin、ERK1/2、c-Myc和c-fos的表达,免疫荧光检测Vimentin的表达;裸鼠皮下注射建立移植瘤模型,30 d后检测瘤重量,绘制存活曲线,检测移植瘤中Vimentin、miR-128-3p、MAPK1、ERK1/2、c-Myc和c-fos的量。 结果　miR-128-3p靶向抑制MAPK1表达;miR-128-3p过表达后,侵袭细胞数目、伤口愈合率降低,E-cadherin表达上调,N-cadherin表达下调,Vimentin阳性率减少,p-ERK1/2、c-Myc和c-fos表达下调。经miR-128-3p干预,裸鼠体内移植瘤重量减轻,存活率增加,miR-128-3p表达上调,MAPK1的表达下调,Vimentin阳性率减少,p-ERK1/2、c-Myc和c-fos表达下调。 结论　过表达miR-128-3p通过靶向抑制MAPK1表达来抑制膀胱癌细胞5637的侵袭能力、迁移能力、上皮-间充质转化和ERK1/2、c-Myc和c-fos通路。     

miR-128-3p过表达靶向MAPK1抑制膀胱癌5637细胞株的侵袭，迁移和上皮间质转化

目的　研究miR-128-3p过表达对膀胱癌5637细胞株的侵袭,迁移和上皮间质转化影响。 方法　基因预测软件TargetScan筛选出miR-128-3p的靶基因,荧光素酶报告实验验证;RT-PCR检测miR-128-3p和MAPK1的表达,Transwell检测细胞侵袭情况,划痕实验检测细胞迁移能力,Western blot检测E-cadherin、N-cadherin、ERK1/2、c-Myc和c-fos的表达,免疫荧光检测Vimentin的表达;裸鼠皮下注射建立移植瘤模型,30 d后检测瘤重量,绘制存活曲线,检测移植瘤中Vimentin、miR-128-3p、MAPK1、ERK1/2、c-Myc和c-fos的量。 结果　miR-128-3p靶向抑制MAPK1表达;miR-128-3p过表达后,侵袭细胞数目、伤口愈合率降低,E-cadherin表达上调,N-cadherin表达下调,Vimentin阳性率减少,p-ERK1/2、c-Myc和c-fos表达下调。经miR-128-3p干预,裸鼠体内移植瘤重量减轻,存活率增加,miR-128-3p表达上调,MAPK1的表达下调,Vimentin阳性率减少,p-ERK1/2、c-Myc和c-fos表达下调。 结论　过表达miR-128-3p通过靶向抑制MAPK1表达来抑制膀胱癌细胞5637的侵袭能力、迁移能力、上皮-间充质转化和ERK1/2、c-Myc和c-fos通路。     

MiR-199-3p replacement affects E-cadherin expression through Notch1 targeting in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) represents the second cause of cancer-related mortality worldwide and is associated with poor prognosis, due to a high recurrence rate after curative treatments and a drug resistance phenotype. In this scenario, the identification of innovative and effective therapeutic strategies is an unmet clinical need. The safety and efficacy of microRNA (miRNA) mediated approaches in preclinical models and clinical trials have been widely described in cancer. MicroRNA-199a downregulation is a common feature of HCC where its reduced expression contributes to mTOR and c-Met pathways activation. Notch1 activation is also a common event in HCC, influencing epithelial-to-mesenchymal transition, tumor invasion and recurrence at least in part through E-cadherin regulation. Here we identified a negative correlation between miR-199a-3p and Notch1 or E-cadherin protein levels in HCC patients and demonstrated that miR-199a-3p regulates E-cadherin expression through Notch1 direct targeting in in vitro models. Moreover, we showed that a strong correlation exists between miR-199a-5p and miR-199a-3p in HCC specimens and that miR-199a-5p contributes to E-cadherin regulation as well, underlying the complex network of interaction carried out by miR-199a and its influence on tumor aggressiveness. In conclusion, our findings suggest the restoration of miR-199a-3p physiologic levels as a possible therapeutic strategy for the treatment of HCC.     

MiR-223-3p overexpression inhibits cell proliferation and migration by regulating inflammation-associated cytokines in glioblastomas

Glioblastoma(GBM) is most common brain tumor in adults. Currently standard treatments have limited effect to increase the survival, because there are still largely unclear mechanisms in glioblastoma development. miR-223 was involved in various types of cancer, however, the function of miR-223-3p in GBM was still unclear. In our study, real-time PCR was performed to exam the expression level of miR-223-3p and NLRP3 (Nucleotide-binding oligomerization domain(NOD)-like receptor family PYRIN domain containing-3) in GBM tissues. Following that, mimic or inhibitor of miR-223-3p were used to modulate miR-223-3p expression in GBM cell lines respectively. Then, we analyzed cell proliferation and migration by cell counting kit and transwell assay. Further, western blot was performed to detect several inflammation-associated cytokines level in GBM cell lines. We found that miR-223-3p was decreased but NLRP3 was increased in GBM tissues. Treatment with miR-223-3p mimic inhibits cell proliferation and migration via decreasing several inflammation-associated cytokines, including interleukin-1β (IL-1β), monocyte chemoattractant protein-1 (MCP-1), IL-8 and IL-18. Importantly, these effects induced by miR-223-3p could be attenuated by NLRP3 overexpression, which was considered as one of target genes of miR-223-3p. In conclusion, these results indicated that miR-223-3p might act as a suppressor and a potential therapy target of GBM.     

Blocking lncRNA MIR155HG/miR-155-5p/-3p inhibits proliferation,invasion and migration of clear cell renal cell carcinoma

This study aimed to investigate the effect of blocking the MIR155HG/miR-155-5p/-3p axis on proliferation, invasion and migration of clear cell renal cell carcinoma. RT-qPCR was used to detect the expression of MIR155HG, miR-155-5p, miR-155-3p in clear cell renal cell carcinoma cell lines. To study the effects of blocking LncRNA MIR155HG and interfering with miR-155-5p and miR-155-3p on the biological function. The g proliferation of tumor was detected by CCK-8, and the cell invasion and migration abilities were detected by wound healing and transwell experiments. Western blot analyzed protein levels of KI67, PCNA, MMP2 and MMP9. Furthermore, TargetScan and miRDB were used to predict the co-target gene of miR-155-3p and miR-155-5p, and the functional analysis of co-target genes was performed using the DAVID. In the current research, the expression of MIR155HG was increased in ccRCC. Interference of MIR155HG inhibited the cellular functions of ccRCC cells, which was reversed by overexpression of miR-155-3p and miR-155-5p.In addition, MIR155HG interference repressed the expression of miR-155-5p and miR-155-3p in ccRCCs, while inhibition of miR-155-5p and miR-155-3p restrained the proliferation, invasion and migration of ccRCCs. Bioinformatics software analysis showed 13 co-targeting genes of miR-155-3p and miR-155-5p. Functional analysis presented that the target genes of miR-31-3p were involved in numerous of biochemical processes and pathways.Blocking lncRNA MIR155HG/miR-155-5p/-3p inhibits proliferation, invasion and migration of renal clear cell carcinoma, which provided a new method for early diagnosis and precise treatment of ccRCC.     

MiR-26b inhibits hepatocellular carcinoma cell proliferation,migration, and invasion by targeting EphA2

Deregulated microRNAs (miRNAs) have been shown to play important roles in cancer progression as a result of changes in expression of their target genes. In this study, we investigated the expression of miR-16b in eight hepatocellular carcinoma (HCC) cell lines, revealed the roles of miR-26b on hepatocellular carcinoma (HCC) cell proliferation, migration, and invasion, and confirmed that EphA2 is a direct target of miR-26b. The miR-26b expression was decreased and EphA2 expression was evaluated in HCC cell lines. Luciferase assays revealed that miR-26b inhibited EphA2 expression by targeting the 3’-untranslated region of EphA2 mRNA. Overexpression of miR-26b dramatically inhibited the proliferation, invasion, and migration of HCC cells by targeting EphA2. Moreover, miR-26b down-regulated c-Myc and CyclinD1 expression, which was reversed by overexpressed EphA2. Taken together, our data demonstrated the mechanism of miR-26b contributed to HCC progression and implicated that miR-26b’s potential in HCC therapy.     

miR-129-5p靶向调控VCP抑制骨肉瘤细胞体外迁徙侵袭

目的 探讨miR-129-5p是否靶向调控VCP基因抑制骨肉瘤细胞迁徙侵袭.方法 构建miR-129-5p过表达及低表达的慢病毒载体,转染骨肉瘤细胞U2-OS;采用实时荧光定量PCR检测上调及下调miR-129-5p的U2-OS细胞中miR-129-5p的表达量;采用RT-PCR和Western blot技术检测VCP mRNA和蛋白表达;采用划痕实验和Transwell侵袭实验检测细胞迁徙、侵袭情况.结果 实时荧光定量PCR结果显示U2-OS细胞中miR-129-5p表达被明显上调或下调;RT-PCR和Western blot检测结果显示:miR-129-5p上调U2-OS细胞组中VCP mRNA和蛋白表达水平显著低于阴性对照细胞组(阴性慢病毒转染);miR-129-5p下调U2-OS细胞组中VCP mRNA和蛋白表达水平显著高于阴性对照细胞组;miR-129-5p上调U2-OS细胞迁徙和侵袭力显著低于阴性对照细胞,miR-129-5p下调的U2-OS细胞迁徙和侵袭力显著高于阴性对照细胞.结论 miR-129-5p靶向调控VCP的表达而抑制骨肉瘤细胞迁徙和侵袭能力.