MiR-21-5p,miR-34a,and human telomerase RNA component as surrogate markers for cervical cancer progression

Objective

This study aimed to demonstrate the predictive value of miR-21-5p, miR-34a, and human telomerase RNA component (hTERC) in cervical cancer (CC) development and evaluated their potential possibility for future clinical applications.

Aberrant expression of microRNA-26b and its prognostic potential in human cervical cancer

Background: microRNA-26b (miR-26b) is reported to be downregulated in many human malignancies and function as a tumor suppressor. However, the roles of miR-26b expression in cervical cancer progression are unclear. The aim of this study was to investigate the clinicopathological or prognostic significance of miR-26b in human cervical cancer. Methods: A cohort of 88 paired of cervical cancer and the adjacent normal cervical epithelial tissues were collected. Quantitative RT-PCR (qRT-PCR) assay was used to detect the expression of miR-26b and its correlations with clinicopathological factors were statistically analyzed. Finally, the survival was assessed by the Kaplan-Meier method and proportional hazards model. Results: The expression level of miR-26b in cervical cancer tissues was significantly lower than that in the adjacent normal cervical tissues (P<0.001). Reduced miR-26b was observed to be significantly correlated with advanced FIGO stage, higher incidence of lymph node metastasis and recurrence of cervical cancer patients (P=0.002, 0.036 and 0.029, respectively). In addition, patients with low-miR-26b expression showed poorer recurrence-free survival (RFS) and overall survival (OS) than those with high-miR-26b expression (P=0.0043 and 0.0015, respectively). Furthermore, multivariate analyses demonstrated that low miR-26b expression was an independent prognostic factor for predicting the 5-year RFS and OS of cervical cancer patients (P=0.013 and 0.007, respectively). Conclusion: Our results showed that reduced miR-26b was correlated with tumor development and poor prognosis in human cervical cancer. The status of miR-26b expression may be a potential prognostic biomarker for cervical cancer patients.     

miR-187在宫颈癌中的表达与临床病理关系的研究

目的 探讨miR-187在宫颈癌中的表达及与临床病理特征的关系。方法 收集78例宫颈癌组织标本,同时收取同期手术的60例正常宫颈组织标本,采用实时荧光定量PCR检测方法检测两种组织中miR-187的表达量并进行比较,同时分析癌组织中miR-187的表达水平与临床病理特征的关系。结果 宫颈癌组织中miR-187的表达量低于正常宫颈组织标本,统计学意义显著（P＜0.001）;宫颈癌组织中miR-187的表达水平与宫颈癌组织的分化程度、患者的临床分期及有无淋巴结转移情况有关（P＜0.05）,而与患者的年龄、肿块类型、肿瘤大小及组织学类型无关（P＞0.05）。结论 miR-187在宫颈癌组织中低表达,可能作为其诊断与治疗的重要靶点。     

Wnt5A expression is associated with the tumor metastasis and clinical survival in cervical cancer

Aims: To identify the clinical significance of Wnt5A expression in the development and progression of cervical cancer. Methods: Real-time PCR was performed in 8 pairs of surgically resected cervical cancer and adjacent normal cervical tissues. Immunohistochemistry was performed to examine Wnt5A expression in 94 paraffin-embedded cervical cancer samples. Associatio ns of Wnt5A expression with clinicopathological factors and clinical survival were analyzed. Results: Wnt5A expression was overexpressed in cervical cancer tissues compared with adjacent normal cervix. Wnt5A expression tended to be positively correlated with lymph nodes metastasis (P = 0.028) and recurrence (P = 0.009). Moreover, patients with higher Wnt5A expression in cancer tissues had better overall (P = 0.004) and recurrent-free survival (P = 0.012) than those with lower Wnt5A expression. Multivariate analysis revealed that Wnt5A was an independent prognostic factor (P = 0.026) for predicting overall survival of cervical cancer patients. Conclusion: Upregulation of Wnt5A was associated with metastasis and progression of cervical cancer. The results of our study unravel the significance of Wnt/Ca2+ signaling in cervical cancer.     

Lgr5 is a potential prognostic marker in patients with cervical carcinoma

Objective: To investigate the Lgr5 (Leucine-rich repeat-containing G protein-coupled receptor 5) expression in cervical carcinoma and to estimate its clinical significance. Methods: The expression of Lgr5 mRNA was evaluated by Real-time PCR in 8 pairs of surgically removed cervical cancer and adjacent normal cervical tissues. Lgr5 protein expression was evaluated by immunohistochemistry in 94 paraffin-embedded cervical carcinoma specimens. The correlation between Lgr5 expression and clinicopathological features were statistically analyzed. Results: Lgr5 expression was significantly higher in cervical cancer tissues compared with that in adjacent normal cervix. High Lgr5 expression was positively correlated with tumor size (P = 0.025) and parametrial infiltration (P = 0.027). Moreover, high levels of Lgr5 was associated with lower overall survival (P = 0.021) and recurrent-free survival (P = 0.008), especially in stage II patients (P = 0.035). Multivariate analysis showed that the expression of Lgr5 was an independent factor of recurrent-free survival for the patients with cervical carcinoma (P = 0.135). Conclusion: Lgr5 may play an important role in the development and progression of cervical carcinoma, and may be a potential therapeutic target for the treatment of cervical carcinoma.     

Detection of circulating exosomal miR-17-5p serves as a novel non-invasive diagnostic marker for non-small cell lung cancer patients

Exosome-shuttled bioactive miRNAs act as novel non-invasive biomarkers for cancer diagnosis have received increasing attention. In this study, we aimed to investigate the expression signatures of exosomal miRNAs and develop a serum exosome-derived miRNA panel for diagnosis of non-small cell lung cancer (NSCLC). The miR-17-92 cluster including 6 miRNAs (miR-17-5p, miR-18a-5p, miR-19a-3p, miR-19b-1-5p, miR-20a-5p and miR-92a-1-5p) was selected as potential diagnostic candidate molecule. Then, expression profiles of the candidate miRNAs were firstly analyzed in 43 pairs of serum samples from the training set by quantitative real-time PCR, and the dysregulated miRNA along with three tumor markers (carcinoembryonic antigen, CEA; cytokeratin 19 fragment, CYFRA21-1; squamous cell carcinoma antigen, SCCA) were further validated in two independent cohorts, which consisted of training set (including 100 NSCLC patients and 90 healthy controls) and validation set (including 72 NSCLC patients and 47 healthy controls). The expression of miR-17-5p was significantly up-regulated in NSCLC patients compared with the healthy controls (P < 0.001), suggesting that miR-17-5p might have considerable clinical value in the diagnosis of NSCLC. Based on the data from the training set, we next used a logistic regression model to construct a 4-molecule panel consisting of miR-17-5p and three tumor markers for NSCLC diagnosis. The performance of such 4-molecule panel was verified with an area under the ROC curve of 0.860 (95% CI = 0.802 to 0.906, sensitivity = 63.0% and specificity = 93.3%) and 0.844 (95% CI = 0.766 to 0.904, sensitivity = 76.4% and specificity = 76.6%) in the training set and validation set, respectively. In conclusion, the newly developed diagnostic panel consisting of exosomal miR-17-5p, CEA, CYFRA21-1 and SCCA may have considerable clinical value in the diagnosis of NSCLC.     

卵巢癌患者miR-497、miR-125a-5p的表达及与其临床特征和生存率的关系

目的:分析卵巢癌患者癌组织中微小RNA-497(miR-497)和微小RNA-125a-5p(miR-125a-5p)的表达情况及其临床意义.方法:选取2018-06—2019-12在本院手术治疗的96例卵巢癌患者作为研究对象,将手术切除的卵巢癌组织作为试验组,癌旁(>2cm)正常组织为对照组.采用实时荧光定量PCR(...     

Linc00152靶向miR-376c-3p对宫颈癌细胞活力、凋亡和放射敏感性的影响

目的:探讨长链非编码RNA Linc00152对宫颈癌细胞活力、凋亡和放射敏感性的影响及作用机制。方法:RT-qPCR检测宫颈癌HeLa细胞和SiHa细胞以及正常宫颈细胞Ect1/E6E7中Linc00152与微小RNA-376c-3p(miR-376c-3p)的表达水平。建立Linc00152低表达或miR-376c-3p过表达的宫颈癌HeLa细胞系,MTT法、流式细胞术、集落形成实验和Western blot分别检测Linc00152低表达或miR-376c-3p过表达对宫颈癌HeLa细胞的活力、凋亡、放射敏感性及相关蛋白表达的影响。双萤光素酶报告基因实验验证Linc00152与miR-376c-3p的调控关系。结果:与Ect1/E6E7细胞相比,宫颈癌HeLa细胞和SiHa细胞中的Linc00152表达上调,miR-376c-3p表达下调(P<0.05)。Linc00152低表达或miR-376c-3p过表达均可抑制HeLa细胞的活力,并诱导其凋亡,增强其放射敏感性,抑制cyclin D和Bcl-2蛋白表达,促进P21和Bax蛋白表达(P<0.05)。Linc00152在HeLa细胞中负向调控miR-376c-3p的表达,抑制miR-376c-3p表达可逆转Linc00152低表达对HeLa细胞活力、凋亡和放射敏感性的作用。结论:宫颈癌细胞中Linc00152高表达;Linc00152通过靶向调控miR-376c-3p影响HeLa细胞的生长、凋亡和放射敏感性,是宫颈癌潜在的诊治靶点。     

miR-21表达异常与乳腺癌临床病理特征及预后的关系

目的： 应用荧光定量PCR技术研究miR-21表达量与乳腺癌临床病理特征及患者预后的关系。方法: 搜集具有5年以上随访资料的乳腺癌病例113例,从甲醛固定、石蜡包埋（FFPE）乳腺癌(BRCA)及其癌旁组织(NATs)中提取总RNA,应用荧光定量RT-PCR技术检测miR-21在乳腺癌及其癌旁组织中的表达量。结果: 与相应癌旁组织比较,miR-21在乳腺癌组织中表达显著上调（P<0.01）,平均上调倍数为1.74 ± 0.48。miR-21表达上调水平与乳腺癌临床分期(P<0.01)、淋巴结转移（P<0.01)及预后［hazard ratio (HR)=5.476, P<0.01］相关。 Cox多重回归分析显示miR-21相对表达量(HR=4.133, P<0.01)为乳腺癌的独立预后因素之一。结论: miR-21表达上调与乳腺癌患者预后不良有关,其可能是一种潜在的乳腺癌独立预后指标。     

MiR-142-3p functions as a tumor suppressor by targeting CD133, ABCG2, and Lgr5 in colon cancer cells

Studies have shown that the expression of CD133, leucine-rich-repeat-containing G-protein-coupled receptor 5 (Lgr5), and ATP binding cassette (ABC)G2 proteins is associated with malignancy and poor prognosis in colon cancer. However, molecular regulation mechanism of the three proteins has not been elucidated. Here, we report that microRNA-142-3p (miR-142-3p) inhibits the expression of CD133, Lgr5, and ABCG2 in colon cancer cells by binding to both the 3′-untranslated region and the coding sequences of the three genes. The miR-142-3p was markedly decreased in colon cancer specimens, in which it was negatively correlated with the expression of CD133, Lgr5, and ABCG2. Reduction of miR-142-3p corresponds to poor differentiation and bigger tumor size in colon cancers. Moreover, miR-142-3p levels were reduced in cells that formed spheres compared to cells that were cultured in regular media. Transfection of miR-142-3p mimics in colon cancer cells downregulated cyclin D1 expression, induced G₁ phase cell cycle arrest, and elevated the sensitivity of the cells to 5-fluorouracil. Furthermore, OCT4 suppressed miR-142-3p, and hypomethylation of the OCT4 promoter was associated with a reduction in miR-142-3p. Finally, the miR-142-3p inhibited the growth of colon cancer cells in vivo, which was accompanied by the downregulation of CD133, Lgr5, and ABCG2 in tumor tissues. Our results elucidate a novel regulation pathway in colon cancer cells and suggest a potential therapeutic approach for colon cancer therapy.     

微小RNA-513c-5p在宫颈癌中的表达及靶向组蛋白去乙酰化酶1调节宫颈癌细胞迁移和侵袭的机制

目的 探讨微小RNA（miR)-513c-5p在宫颈癌中的表达及靶向组蛋白去乙酰化酶1（HDAC1）调节宫颈癌细胞迁移和侵袭的机制。方法 临床收集宫颈癌患者86例,通过Real-time PCR检测肿瘤组织和癌旁组织中miR-513c-5p水平,分析其与宫颈癌病理特征的关系。通过双荧光素酶报告验证miR-513c-5p靶向HDAC1。将宫颈癌HeLa细胞系分为4组：对照组、类似物(mimic)组、mimic+HDAC1组和HDAC1组。通过质粒转染技术过表达miR-513c-5p和(或)HDAC1。Real-time PCR和Western blotting分别用于检测RNA或蛋白的表达水平。分别通过CCK-8法、细胞划痕实验和Transwell实验检测各组的细胞生长、迁移和侵袭能力。 结果 宫颈癌组织中miR-513c-5p水平显著低于癌旁组织。低水平的miR-513c-5p与更高的局部侵袭、淋巴转移和远端转移有关（P<0.05）。miR-513-5p靶向抑制HDAC1表达。过表达miR-513c-5p显著抑制宫颈癌细胞生长、迁移和侵袭（P<0.05）。过表达HDAC1促进细胞生长、迁移和侵袭（P<0.05）,并且可以逆转miR-513c-5p的抑制作用（P<0.05）。 结论 低水平的miR-513c-5p可能与宫颈癌转移有关,并且miR-513c-5p可通过靶向抑制HDAC1蛋白的表达抑制宫颈癌HeLa细胞生长、迁移和侵袭。     

MicroRNA-153 expression and prognosis in non-small cell lung cancer

Background: miR-153 has been found to be significantly decreased in non-small cell lung cancer (NSCLC) tissues; however, its clinical significance has not been investigated. Methods: The expression patterns of miR-153 in 137 pairs of human lung cancer tissues and adjacent normal lung tissues were analyzed using qRT-PCR. The relationships between miR-153 expression and clinicopathological parameters were examined by chi-square test. Kaplan-Meier method and the log-rank test were used to determine the difference in overall survival (OS) rates between two groups. Results: The expression of miR-153 was reduced significantly, compared with adjacent normal lung tissues (P<0.05). We observed that the expression level of miR-153 was positively correlated with the clinical stage (P=0.005), lymph node status (P=0.014), distant metastasis (P=0.004), and differentiated degree (P<0.001) in NSCLC patients. According to the Kaplan-Meier survival analysis, the patients with low miR-153 expression exhibited evidently poorer overall survival rates than those with high miR-153 expression (P=0.003). Multivariate analysis showed that the expression of miR-153 was an independent and significant factor associated with poor OS rates (P=0.002). Conclusion: Decreased expression of miR-153 might be a potential unfavorable prognostic factor for patients with NSCLC, and further studies would be needed to prove our findings.     

头颈部鳞状细胞癌预后相关的miRNAs的生物信息学分析

目的 利用癌症基因组图谱（TCGA）数据库微小核糖核酸（miRNAs）表达谱数据分析头颈部鳞状细胞癌（HNSCC）与癌旁正常组织间差异表达的miRNAs,结合临床信息寻找与HNSCC预后相关的miRNAs。方法 从TCGA中下载miRNAs表达数据,包括39例HNSCC患者和39个肿瘤邻近正常组织样本筛选差异表达的miRNAs,应用481例HNSCC患者的miRNAs表达谱和临床信息来评估找到的差异表达miRNAs的预后作用。结果 共筛选出114个差异表达的miRNAs,包括60个上调和54个下调的miRNAs。Kaplan-Meier生存分析显示miR-4652-5p和miR-99a-3p与HNSCC患者预后相关,单因素和多因素Cox回归分析显示,miR-4652-5p和miR-99a-3p是HNSCC的重要预后因素。结论 miR-4652-5p和miR-99a-3p与HNSCC患者预后相关,但miR-4652-5p和miR-99a-3p在头颈鳞状细胞癌发生发展中的分子机制仍需更全面的基础和临床研究进行探讨。     

MicroRNA-490-5p is a novel tumor suppressor targeting c-FOS in human bladder cancer

Introduction

Recent studies have demonstrated the critical roles of micro-RNAs in tumorigenesis and tumor progression. Here, we describe the regulation and function of miR-490-5p in bladder cancer.

Material and methods

Paired tissue samples were collected from bladder cancer patients (n = 20). Real-time PCR revealed that miR-490-5p expression was significantly down-regulated in human bladder cancer tissues and cells. Also there was an inverse relationship between the expression level of miR-490-5p and the pathological grade of bladder cancer. Western blotting was performed to detect the expression levels of c-FOS and TET1 in 6 matched tumor tissue samples and 4 bladder cell lines. Furthermore, to better understand the underlying mechanisms of miR-490-5p, we conducted gain and loss of function analysis by transfecting bladder cancer T24 cells with chemically synthesized miR-490-5p mimics and inhibitor, respectively.

Results

We found that overexpression of miR-490-5p in T24 cells could inhibit cell proliferation and invasion and induce cell apoptosis. Conversely, suppression of miR-490-5p expression induced cell proliferation and invasion, while it inhibited cell apoptosis. In addition, our bioinformatics prediction and experimental data showed that c-FOS was a potential target of miR-490-5p. The expression level of c-FOS was significantly decreased after miR-490-5p overexpression and significantly increased after miR-490-5p suppression, indicating that c-FOS was a target of miR-490-5p.

Conclusions

These findings suggest that miR-490-5p is a novel tumor suppressor, contributing to the carcinogenesis of bladder cancer by targeting c-FOS.     

Clinicopathologic and prognostic significance of p21 (Cip1/Waf1) expression in bladder cancer

Recent studies have shown that altered expression p21 is shown to associate with tumorigenesis and tumor progression. To investigate the clinicopathological significance and prognostic value of p21 in bladder cancer (BCa). A total of 48 patients with BCa were included in this study. The correlation between p21 expression and clinicopathologic features and survival was studied. Also, a meta-analysis was performed to investigate the relationship between the p21 and BCa survival. Low p21 expression was detected both in tumor tissues compared with adjacent normal tissues. The expression of p21 was closely associated with advanced pathologic TNM stage (P = 0.001) and tumor grade (P = 0.013). Moreover, patients with low p21 expression had shorter recurrence-free survival (P = 0.016) and overall survival rates (P = 0.039). Multivariate Cox regression analysis revealed that p21 low expression was an independent prognostic factor for recurrence free survival (P = 0.03). Additionally, our meta-analysis. The available outcome data from six articles were examined. A meta-analysis of the HR indicated a significantly poor overall survival (OS, HR: 1.75, 95% CI: 1.38-2.21), recurrence free survival (RFS, HR: 1.83, 95% CI: 1.57-2.15), progression free survival (PFS, HR: 2.02, 95% CI: 1.48-2.75), and cancer specific survival (CSS, HR: 1.89, 95% CI: 1.53-2.33) in patients with low expression levels of p21. Our present results indicated that low p21 expression predicated tumor recurrence and poor prognosis in bladder cancer.     

MiR-145-5p在大肠癌中的表达及与多药耐药的关系

目的 探讨大肠癌中miR-145-5p和多药耐药基因(MDR1)蛋白P-糖蛋白（P-gp）的表达及两者的关系。方法 分别在组织、细胞水平采用SP法、Real-time PCR法、Western blotting法检测50例大肠癌 (CRC)患者组织及30例癌旁正常组织患者中P-gp的表达,miR-145-5p的表达变化对MDR1 mRNA及P-gp的影响及两者与临床病理特征之间的相关性。结果 大肠癌组织中miR-145-5p表达量明显低于癌旁正常组织,MDR1 mRNA及P-gp的表达量明显高于癌旁正常组织(r=-0.403,P<0.01)。大肠癌细胞(HCT-15)中miR-145-5p 能够抑制MDR1 mRNA及P-gp的表达(P<0.05)。结论 MiR-145-5p对大肠癌多药耐药基因及蛋白的表达具有调控作用,参与了大肠癌的发生、发展及多药耐药。     

Decreased expression of serum miR-424 correlates with poor prognosis of patients with hepatocellular carcinoma

Background: MicroRNAs (miRNAs) play important roles in many important cellular processes and deregulation of miRNAs is linked to many human diseases including cancer. Although miR-424 has been demonstrated to inhibit progression of hepatocellular carcinoma (HCC), its expression level in serum samples and the potential clinical values remain unknown. Materials and methods: The expression level of miR-424 in the serum clinical samples from HCC patients and healthy volunteers were determined by qRT-PCR. Then the association of serum miR-424 expression level with various important clinicopathological parameters and survival rates was evaluated. Multivariate Cox regression analysis was used to identify the independent risk factors for HCC. Results: The expression level of serum miR-424 was significantly decreased in patients with HCC compared with the healthy volunteers (P<0.01). Reduced expression of serum miR-424 was associated with serum AFP (P=0.048), vein invasion (P=0.006) and TNM stage (P=0.003). In addition, survival analysis showed that HCC patients with lower serum miR-424 expression suffered poorer overall survival (P=0.018) and disease free survival (P=0.008). Moreover, serum miR-424 was demonstrated to be an independent risk factor for HCC. Conclusions: Our findings provide the compelling evidence that the decreased expression of serum miR-424 may serve as a novel biomarker to predict the unfavorable prognosis of HCC patients.     

子宫颈癌中miR-153和Survivin的表达及其临床意义

目的探讨miR-153和Survivin在子宫颈癌组织中的表达及其对子宫颈癌的临床意义。方法收集50例子宫颈癌和40例正常子宫颈组织,采用RT-PCR法和免疫组化SP法检测miR-153和Survivin在子宫颈癌和正常子宫颈组织中的表达,分析两者表达与子宫颈癌临床病理特征的关系及对患者预后的意义。结果miR-153在子宫颈癌组织中的相对表达量低于正常子宫颈组织,Survivin在子宫颈癌组织中的相对表达量高于正常子宫颈组织,差异均有统计学意义(t=16.333,t=6.407,P<0.01)。通过散点图及相关性分析,在子宫颈癌组织中miR-153和Survivin相对表达量呈负相关,差异有统计学意义(P<0.01);miR-153、Survivin表达与患者年龄、肿瘤大小、TNM分期均无关(P>0.05),与淋巴结转移明显相关(χ^2=6.455,χ^2=5.937,P<0.05)。通过散点图及相关性分析,淋巴结转移患者的淋巴结受侵数目与miR-153表达量呈负相关,与Survivin表达量呈正相关,差异均有统计学意义(P<0.01)。预后生存分析发现,miR-153高表达患者生存率高于低表达患者,Survivin高表达患者生存率低于低表达患者,差异均有统计学意义(χ^2=19.390,χ^2=27.921,P<0.01)。结论miR-153在子宫颈癌组织中低表达,与Survivin在子宫颈癌组织中表达水平呈负相关,可能与子宫颈癌的增殖、侵袭转移有关。     

miR-155-5p在宫颈癌血清中的表达及其对宫颈癌细胞的影响

 目的：检测miR-155-5p在不同宫颈疾病患者血清中的表达差异,并分析其对宫颈癌细胞增殖、细胞周期和凋亡的影响,探讨miR-155-5p在宫颈癌发生、发展中的可能作用机制。方法：采用SYBR GreenⅠ实时荧光定量PCR法,检测并分析比较miR-155-5p在不同宫颈疾病患者血清中的表达差异。利用miR-155-5p mimic或inhibitor提高或降低宫颈癌细胞中miR-155-5p的表达。CCK-8法和流式细胞术检测宫颈癌细胞的增殖、细胞周期和凋亡。结果：宫颈癌组血清中miR-155-5p的表达高于宫颈炎组和健康对照组（P<0.05）,宫颈上皮内瘤样病变组和宫颈癌组血清中miR-155-5p的表达差异无统计学意义（P>0.05）。与空白组、脂质体组和阴性对照组相比,转染100 nmol/L和200 nmol/L miR-155-5p mimic的SiHa细胞中,S期细胞比例升高,凋亡细胞比例降低（P<005）。转染100 nmol/L和200 nmol/L miR-155-5p inhibitor的SiHa细胞中,G2/M期细胞比例明显增多（P<005）。结论：(1)宫颈癌患者血清中miR-155-5p表达较健康对照人群上调,可能作为宫颈癌早期诊断的肿瘤分子标志物。(2)miR-155-5p对宫颈癌HeLa细胞增殖、细胞周期和凋亡无明显影响。(3)miR-155-5p可促进宫颈癌SiHa细胞进入S期,并抑制SiHa细胞凋亡,提示miR-155-5p可能在宫颈鳞癌发生、发展中起作用。