Analysis of miR-205 and miR-155 expression in the blood of breast cancer patients

The purpose of this study was to identify and validate circulating microRNAs (miRNAs) in human plasma for use as breast cancer (BC) biomarkers and to analyze their relationship to clinicopathologic features and its preliminary biological function. Genome-wide expression profiling of miRNAs in BC was investigated by microarray analysis. miR-155 was up-regulated greater than two-fold in BC compared with Normal Adjacent Tissue (NAT), whereas let-7b, miR-381, miR-10b, miR-125a-5p, miR-335, miR-205 and miR-145 were down- regulated greater than two-fold. Our hypothesis was that circulating miRNAs are also present and differentially expressed in the serum of BC patients compared to controls. Using real-time PCR (RT-PCR), we analyzed miR-205 and miR-155 in archived serum from 30 participants, 20 with breast cancer and 10 healthy people. miR-205 was down-regulated in BC patient serum while miR-155 was up-regulated. Furthermore, we analyzed the relationship between the expression levels of these two miRNAs and the clinicopathologic parameters of BC patients. High expression of miR155 was associated with clinical stage, molecular type, Ki-67 and p53 in BC patients (P<0.05). By contrast, we found no significant correlation between miR-205 and BC patient clinicopathologic parameters. Functional analysis showed that ectopic expression of miR-205 significantly inhibits cell proliferation and promotes apoptosis. miR-205 was down-regulated and miR-155 was up-regulated in BC patient serum. miR-155 was positive correlated with clinical stage and ki-67 and negatively correlated with p53 status.     

Association between miR-196a-2 Gene Polymorphism and Ovarian Cancer Prognosis in Egyptian Females

Background: Ovarian cancer is the fifth leading cause of cancer-related deaths among women worldwide. Unfortunately, early detection tests are relatively lacking. Diagnosis in the late stages of the disease carries a poor prognosis. Objective: To evaluate the relationship between miR-196a-2 rs11614913 polymorphism and ovarian cancer risk and prognosis in Egyptian females. Methods: In this case-control study, the participants were classified into 2 groups. Group A is the control group which included 50 healthy females. Group B included 50 patients newly diagnosed with ovarian carcinoma confirmed by histopathological analysis. Immunohistochemistry for P53 and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for miR-196a-2 genotypes detection were performed.   Results: There was a statistically significant difference among ovarian cancer cases and controls regarding genotypes (P = 0.003). However, the distribution of the T and C alleles in both studied groups showed no significant difference (P = 0.17). There was a statistically significant increase of CA 125 levels among CT and CC genotypes carriers of ovarian cancer cases (p = 0.04). Besides, there was a statistically significant correlation between miR-196a-2 polymorphism and each of tumor grade (P <0.001), p53 immunohistochemical expression (P= 0.002), and Figo classification (P <0.001). Conclusion: There was a statistically significant increase of CA 125 levels among C allele carriers of ovarian cancer cases. Besides, there was a statistically significant association between the miR-196a-2 polymorphism and each of tumor grade, p53 immunohistochemical expression, and Figo classification. So, miR-196a-2 polymorphism can be a possible prognostic factor in ovarian cancer.     

miR-3666在胃癌中的表达及其临床意义

目的:检测胃癌组织中miR-3666的表达并分析其与临床病理特征的关系。方法:采用茎环逆转录实时定量PCR技术检测80例胃癌手术标本与相对应的癌旁组织中miR-3666的相对表达量,并分析其与胃癌临床病理参数和预后的关系。结果:miR-3666在胃癌组织中表达与癌旁组织相比显著下调（P<0.001）,miR-3666的表达与胃癌淋巴结转移（P=0.002）和TNM分期（P=0.017）相关,与年龄、性别、浸润深度、肿瘤位置及分化程度无关（P>0.05）。合并淋巴结转移的胃癌组织中miR-3666的表达低于无淋巴结转移的胃癌组织（P<0.001）。miR-3666低表达与高表达的患者1年及3年生存率分别为78.0%、22.6%和96.9%、48.1%,中位生存率分别为16个月和29个月,差异有统计学意义（P<0.001）。COX预后分析发现miR-3666低表达和肿瘤TNM分期是胃癌患者预后的独立影响因素。结论:miR-3666在胃癌组织中的表达量明显降低,提示miR-3666可能参与了胃癌的发生发展,并且与胃癌的淋巴结转移相关;miR-3666低表达提示胃癌患者预后不良。     

Association of miR-193b Down-regulation and miR-196a up-Regulation with Clinicopathological Features and Prognosis in Gastric Cancer

Dysregulated expression of microRNAs (miRNAs) has been shown to be closely associated with tumordevelopment, progression, and carcinogenesis. However, their clinical implications for gastric cancer remainelusive. To investigate the hypothesis that genome-wide alternations of miRNAs differentiate gastric cancer tissuesfrom those matched adjacent non-tumor tissues (ANTTs), miRNA arrays were employed to examine miRNAexpression profiles for the 5-pair discovery stage, and the quantitative real-time polymerase chain reaction (qRTPCR)was applied to validate candidate miRNAs for 48-pair validation stage. Furthermore, the relationshipbetween altered miRNA and clinicopathological features and prognosis of gastric cancer was explored. Amonga total of 1,146 miRNAs analyzed, 16 miRNAs were found to be significantly different expressed in tissues fromgastric cancer compared to ANTTs (p<0.05). qRT-PCR further confirmed the variation in expression of miR-193band miR-196a in the validation stage. Down-expression of miR-193b was significantly correlated with Laurentype, differentiation, UICC stage, invasion, and metastasis of gastric cancer (p<0.05), while over-expression ofmiR-196a was significantly associated with poor differentiation (p=0.022). Moreover, binary logistic regressionanalysis demonstrated that the UICC stage was a significant risk factor for down-expression of miR-193b (adjustedOR=8.69; 95%CI=1.06-56.91; p=0.043). Additionally, Kaplan-Meier survival curves indicated that patientswith a high fold-change of down-regulated miR-193b had a significantly shorter survival time (n=19; mediansurvival=29 months) compared to patients with a low fold-change of down-regulated miR-193b (n=29; mediansurvival=54 months) (p=0.001). Overall survival time of patients with a low fold-change of up-regulated miR-196a (n=27; median survival=52 months) was significantly longer than that of patients with a high fold-changeof up-regulated miR-196a (n=21; median survival=46 months) (p=0.003). Hence, miR-193b and miR-196a maybe applied as novel and promising prognostic markers in gastric cancer.     

Aberrant Expression of miR-20a and miR-203 in Cervical Cancer

MicroRNAs (miRNAs) are small, non-coding RNAs that are critical regulators of various diseases. MicroRNA-20a (miR-20a) and microRNA-203 (miR-203) have previously shown significant alteration in a range of cancers.In this study, the expression levels of miR-20a and miR-203 in 100 cervical cancer tissues were detected byqRT–PCR and compared to patient matched-nontumor cervical tissues. Correlations between expression leveland clinicopathologic characteristics of cervical cancer were also analyzed. Finally, we studied the effect of miR-20a and miR-203 on cell proliferation in cervical cancer cell lines by MTT. We found that the expression levelof miR-20a (P<0.001) was significantly higher in cervical cancer patients than in healthy controls, while thatof miR-203 (P<0.001) was lower. Aberrant expression of miR-20a was correlated with lymph node metastasis(LNM), histological grade and tumor diameter, but down-regulated miR-203 was correlated with LNM only.Furthermore, we found that over-expression of miR-203 decreased cell proliferation, while reduction of miR-20a also prevented tumor progression. Our results support the involvement of miR-20a and miR-203 in cervicaltumorigenesis. We propose that miRNAs might be used as therapeutic agents for cervical cancer.     

miR-335、Survivin表达及对乳腺癌术后患者预后评估的价值

目的:探讨微小RNA335(microRNA-335,miR-335)和Survivin在乳腺癌患者组织中表达情况以及其表达水平对患者预后的影响。方法:收集我院2010年2月1日至2014年2月1日乳腺癌患者标本140例,通过实时免疫荧光定量聚合酶链反应（RT-PCR)法检测所有乳腺癌组织、癌旁组织中miR-335的表达情况;采用免疫组织化学方法检测癌组织、癌旁组织中Survivin的表达情况。应用SPSS 16.0软件对比miR-335、Survivin与乳腺癌患者临床病理之间的关系,以及对患者预后的影响。结果:miR-335在癌组织中的表达明显低于癌旁组织(25.36% vs 82.51%,P=0.003),Survivin在癌组织中的表达明显高于癌旁组织(80.13% vs 26.73%,P=0.001 6),miR-335在乳腺癌组织中的表达水平与肿瘤病理类型、分化程度、临床分期呈负相关（r=-0.47,P=0.02;r=-0.31,P=0.03;r=-0.75,P=0.04),而Survivin在乳腺癌组织中的表达水平与肿瘤病理类型、分化程度、临床分期呈正相关（r=0.52,P=0.03;r=0.63,P=0.01;r=0.37,P=0.03)。COX回归模型发现乳腺癌患者肿瘤TNM分期、淋巴结转移、分化程度、病理类型、ER、PR、Her-2、miR-335、Survivin表达均为影响乳腺癌患者PFS的因素。miR-335对于乳腺癌患者术后3年 PFS、OS预测曲线下面积分别为83.4%、78.6%(P<0.01);Survivin 对于乳腺癌患者术后3年PFS、OS预测曲线下面积分别为79.5%、70.6%(P<0.01)。差异有统计学意义（P<0.01,P<0.01)。结论:miR-335、Survivin在乳腺癌组织和癌旁正常组织中的表达存在明显差异,miR-335高表达、Survivin低表达时,提示乳腺癌患者手术预后良好。     

血清外泌体miRNA 对结直肠癌患者免疫调控及预后价值的研究

目的:研究血清外泌体miRNA 对结直肠癌患者免疫调控及预后价值。方法:本研究采取前瞻性病例对照研究,本次研究对象主要以2016年1月到2017年1月在我院进行诊断并治疗的结直肠癌患者180例作为研究对象,同期收集在我院进行肠镜检查,经病理确诊为大肠腺瘤患者60例以及同期健康体检的志愿者60例作为对照,比较三组患者以及不同疾病进展以及预后患者的miR-21、miR-92、miR-196a和miR-196b水平,研究miR-21、miR-92、miR-196a和miR-196b水平的联合检测对患者的不良预后的诊断价值。结果:结直肠癌组、大肠腺瘤组以及健康组的miR-21（F=18.47,P=0.000）、miR-92（F=16.68,P=0.000）、miR-196a（F=15.04,P=0.000）和miR-196b（F=16.31,P=0.000）水平之间的差异存在统计学意义,miR-21、miR-92、miR-196a和miR-196b水平从高到低依次为结直肠癌组、大肠腺瘤组以及健康组;Ⅲ-Ⅳ期、Ⅰ-Ⅱ期患者的miR-21（t=20.740,P=0.000）、miR-92（t=12.265,P=0.000）、miR-196a（t=4.114,P=0.000）和miR-196b（t=3.449,P=0.000）水平之间的差异存在统计学意义。miR-21、miR-92、miR-196a和miR-196b从高到低依次为Ⅲ-Ⅳ期、Ⅰ-Ⅱ期;通过对患者的随访分析,生存患者112例,死亡患者68例,生存组以及死亡组患者的miR-21（t=20.635,P=0.000）、miR-92（t=17.177,P=0.000）、miR-196a（t=12.974,P=0.000）和miR-196b（t=5.010,P=0.000）水平之间的差异存在统计学意义。miR-21、miR-92、miR-196a和miR-196b从高到低依次为死亡组、生存组;通过联合检测效能进行分析,miR-21、miR-92、miR-196a和miR-196b的联合检测对于结直肠癌组患者的特异度显著高于单独检测,通过ROC曲线分析,对于结直肠癌患者的预测分析,miR-21、miR-92、miR-196a和miR-196b的临界值分别为15.63、9.98、1.77、4.02。结论:血清外泌体miRNA 对结直肠癌患者免疫调控及预后具有重要价值,可作为日后诊断的重要依据。     

结肠癌患者血清中miR-30a、miR-106b的表达及其临床意义

目的:探讨结肠癌患者血清miR-30a、miR-106b的表达及其在结肠癌进展及预测患者预后中的价值。方法:选取2012年06月至2013年02月本院收治的接受结肠癌手术治疗的患者80例为研究对象,20例同期健康人群作为对照组。逆转录 PCR法检测外周血血清中 miR-30a、miR-106b的表达水平,分析患者血清miR-30a、miR-106b表达与结肠癌临床病理及预后的关系。Spearman分析miR-30a、miR-106b表达的相关性。采用多因素logistic回归模型对结肠癌患者的预后影响因素进行分析,采用Kaplan-Meier生存分析miR-30a低表达组、miR-30a高表达组,miR-106b低表达组、miR-106b高表达组患者中位生存时间。结果:结肠癌患者血清miR-30a表达水平（1.03±0.02）低于正常人群（5.15±0.06）,差异具有统计学意义（t=16.38,P=0.01）;结肠癌患者血清miR-106b表达水平（2.62±0.35）高于正常人群（1.15±0.06）,差异具有统计学意义（t=10.17,P=0.03）。miR-30a、miR-106b表达与患者的组织分化类型、浆膜侵犯、淋巴结转移、远处转移、TNM分期有显著相关性(P＜0.05)。多因素logistic回归模型对结肠癌患者预后影响因素进行分析,TNM分期（Ⅲ+Ⅳ）、miR-106b高表达及miR-30a低表达是结肠癌患者预后不良的危险因素。miR-30a低表达组患者总体生存时间（55.3±4.1）个月低于miR-30a高表达组（76.4±2.4）个月（P=0.000）;miR-106b低表达组患者的总体生存时间（75.1±7.3）个月高于miR-106b高表达组（51.8±3.1）个月（P=0.000）。结论:miR-30a在结肠癌患者血清中低表达,miR-106b在结肠癌患者血清中高表达,二者的表达与肿瘤的进展、预后不良有关。     

血清miR-135及miR-601在胃癌患者中的表达及其诊断价值

背景与目的：胃癌是消化内科常见的恶性肿瘤。探讨血清miR-135及miR-601在胃癌患者中的表达及其诊断价值。方法：选取2016年1月1日—2019年9月30日三亚中心医院收治的胃癌患者152例,根据胃癌病情进展及临床病理学分期分为早期胃癌组（n=62）和进展期胃癌组（n=90）,Ⅰ～Ⅱ期（n=65）和Ⅲ～Ⅳ期（n=87）,无淋巴结转移组（n=73）和淋巴结转移组（n=79）。另选择96例非胃癌患者作为非胃癌组,60例健康体检正常者作为对照组。采用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR）检测各组血清miR-135及miR-601表达水平,化学发光法测定糖类抗原72-4（carbohydrate antigen 72-4,CA72-4）及糖类抗原19-9（carbohydrate antigen 19-9,CA19-9）水平,分析其对早期胃癌的诊断价值。采用Pearson相关分析来分析胃癌患者血清miR-135、miR-601水平与CA72-4及CA19-9的相关性。结果：与非胃癌组和对照组比较,胃癌组血清miR-135、miR-601、CA72-4及CA19-9水平均明显升高（P均<0.001）。进展期胃癌组血清miR-135（5.70±1.84 vs 3.83±1.12）、miR-601（11.28±3.73 vs 7.36±2.15）、CA72-4［（41.75±10.14）U/mL vs（17.82±4.93）U/mL］及CA19-9［（63.72±17.50）U/mL vs （35.84±10.36）U/mL］水平均明显高于早期胃癌组（均P<0.001）。Ⅲ～Ⅳ期胃癌患者血清miR-135（6.10±1.90 vs 3.74±1.08）、miR-601（12.14±3.92 vs 7.05±2.04）、CA72-4［（44.68±12.35）U/mL vs（16.40±4.52）U/mL］和CA19-9［（68.53±19.13）U/mL vs（33.75±10.60）U/mL］水平均明显高于Ⅰ～Ⅱ期（均P<0.001）,而且Ⅱ期胃癌患者血清miR-135、miR-601、CA72-4及CA19-9水平均明显高于对照组（P<0.05）。淋巴结转移组血清miR-135（6.24±1.95 vs 3.65±0.97）、miR-601（12.60±4.13 vs 6.84±1.92）、CA72-4［（48.70±12.37）U/mL vs （14.85±4.20）U/mL］和CA19-9［（72.36±20.25）U/mL vs（31.60±10.17）U/mL］水平均明显高于无淋巴结转移组（均P<0.001）。受试者工作特征（receiver operating characteristic,ROC）曲线分析结果显示,血清miR-135、miR-601、CA72-4及CA19-9水平诊断早期胃癌的最佳截断值分别为3.78、7.14、17.63 U/mL、35.70 U/mL,四项联合诊断早期胃癌的 曲线下面积（area under curve,AUC）（0.920,95% CI：0.860～0.978）最大,其灵敏度和特异度分别为98.6%和77.4%。相关分析显示,胃癌患者血清miR-135、miR-601水平与CA72-4及CA19-9均呈正相关（r=0.748,P<0.001,r=0.694,P<0.001;r=0.815,P<0.001;r=0.716,P<0.001）。结论：胃癌患者血清miR-135、miR-601、CA72-4及CA19-9水平明显升高,四项联合检测有助于提高早期胃癌的诊断价值。     

血清miR-424及miR-520在多发性骨髓瘤中的表达及其临床意义

目的:探讨多发性骨髓瘤（multiple myeloma,MM）患者血清miR-424及miR-520的表达及其临床意义。方法:选取2015年1月至2019年12月我院收治的87例MM患者,依据国际分期系统分为Ⅰ期（n=18）、Ⅱ期（n=26）和Ⅲ期（n=43）,根据免疫分型结果分为IgG型（n=49）、IgA型（n=20）、轻链型（n=11）和非分泌型（n=7）。另选取同期60例正常者作为对照组。检测各组血清miR-424、miR-520、胱抑素C（Cys-C）及β2-微球蛋白（β2-MG）水平。应用受试者工作特征（ROC）曲线分析血清miR-424、miR-520、Cys-C及β2-MG水平诊断MM的价值。采用Pearson相关分析MM患者血清miR-424、miR-520水平与Cys-C及β2-MG的相关性。结果:MM组血清miR-424（2.93±1.35 vs 0.82±0.27）、miR-520（2.14±0.86 vs 0.56±0.18）、Cys-C（2.16±0.84 vs 0.75±0.18,mg/L）及β2-MG（6.24±2.37 vs 1.80±0.66,mg/L）水平均明显高于对照组（P＜0.001）。MM患者Ⅲ期血清miR-424（5.20±2.46 vs 2.28±0.95,1.17±0.38）、miR-520（3.60±1.51 vs 1.87±0.72,0.83±0.26）、Cys-C（3.75±1.28 vs 1.64±0.53,0.90±0.21,mg/L）及β2-MG（8.82±3.14 vs 5.25±1.38,3.12±0.96,mg/L）水平均明显高于Ⅰ期和Ⅱ期（P＜0.001）,且Ⅱ期血清miR-424（2.28±0.95 vs 1.17±0.38）、miR-520（1.87±0.72 vs 0.83±0.26）、Cys-C（1.64±0.53 vs 0.90±0.21）及β2-MG（5.25±1.38 vs 3.12±0.96）水平均明显高于Ⅰ期（P＜0.001）。MM患者IgG型血清miR-424（4.37±2.18 vs 2.40±1.06,2.27±0.93,2.35±0.84）及miR-520（3.22±1.37 vs 1.72±0.74,1.68±0.63,1.78±0.65）水平均明显高于IgA型、轻链型和非分泌型（P＜0.001）。ROC曲线分析显示,miR-424、miR-520、Cys-C及β2-MG四项联合诊断MM的曲线下面积（0.963,95%CI:0.910～0.998）最大,其敏感度和特异度为97.0%和92.0%。相关分析结果显示,MM患者血清miR-424及miR-520表达水平与Cys-C及β2-MG均呈正相关（r=0.748,P＜0.001,r=0.862,P＜0.001,r=0.705,P＜0.001,r=0.803,P＜0.001）。结论:MM患者血清miR-424及miR-520表达水平明显升高,且随着MM病情分期进展呈升高趋势,联合Cys-C及β2-MG检测对MM诊断具有较高的价值。     

miR-345在非小细胞肺癌中的表达及临床意义

目的 探讨microRNA-345(miR-345)在非小细胞肺癌(NSCLC)中的表达情况,并进一步分析其与肿瘤的临床病理特征和预后关系。方法 利用Real-time PCR检测98例NSCLC患者组织及三种细胞系中miR-345的表达,分析miR-345的表达与临床预后的关系。结果 和对照组相比,miR-345的表达在NSCLC组织及细胞系中显著下调(P＜0.05);组织中miR-345的表达水平和临床病理指标包括淋巴结转移(P＜0.001)、远端转移(P=0.028)、TNM分级(P=0.004)以及病理分期(P=0.011)有关;低表达miR-345的NSCLC患者组和高表达miR-345的患者组相比,其总生存率较低(P=0.021)。多因素分析表明miR-345的表达是NSCLC预后的独立危险因素(HR=3.897,95% CI:2.263~10.440;P=0.012)。结论 miR-345在NSCLC组织及细胞系中的表达降低,低表达的miR-345和NSCLC的进展及预后有关,表明miR-345可能作为NSCLC的一个潜在临床诊断标记。     

miR-150在宫颈癌组织中的表达及临床意义

目的:探讨宫颈癌组织中miR-150的表达及意义.方法:收集62例宫颈癌组织样本,其中Ⅰb期患者23例,Ⅱa期患者29例、Ⅱb期患者10例,正常宫颈组织13例,采用实时荧光定量PCR方法检测miR-150在不同宫颈组织中的表达,并将检测结果和临床及病理资料进行统计学分析.结果:宫颈癌组织中miR-150mRNA的表达量显著高于相应癌旁组织;宫颈癌组织miR-150mRNA的表达量明显高于正常宫颈组织,差异具有统计学意义(P＜0.01);宫颈癌患者癌组织中miR-150 mRNA的表达水平与宫颈癌的临床分期相关,与宫颈癌Ⅰb期患者相比,Ⅱa期以及Ⅱb期宫颈癌患者癌组织中miR-150 mRNA的表达水平明显增高,差异具有统计学意义(P＜0.05),而宫颈癌Ⅱa期与Ⅱb期患者的miR-150 mRNA表达水平之间无明显差异(P＞0.05).结论:miR-150在宫颈癌组织中高表达,可能作为癌基因调控宫颈癌的发生及发展.     

miR-214在胃癌患者外周血中的表达及其临床意义

目的探讨miR-214在胃癌中的表达及临床意义。方法采用实时荧光定量PCR法,检测40例胃癌患者癌组织及癌旁组织中miR-214的差异表达;同时检测血浆miR-214在56例胃癌患者及40例正常组织中的表达。结果miR-214在胃癌组织中的表达水平明显高于癌旁组织;与正常组织比较,胃癌患者中的血浆miR-214表达明显增高;血浆miR-214表达与患者性别、年龄无关,与疾病分期及生存期密切相关,差异具有统计学意义(P<0.05);Cox回归分析发现疾病分期、miR-214的表达可作为独立的预后因子(P<0.05)。结论循环miR-214表达与胃癌肿瘤分期和生存时间相关,检测循环miR-214可能作为评估胃癌患者临床预后的靶基因。     

宫颈癌患者血清miR-196a的表达及其与SCC-Ag及CEA的相关性研究

目的:探讨宫颈癌患者血清miR-196a的表达与鳞状细胞癌抗原（SCC-Ag）及癌胚抗原（CEA）的相关性,分析血清miR-196a、SCC-Ag及CEA水平对宫颈癌的诊断价值。方法: 选取2016年1月至2019年3月海南妇产科医院收治的158例宫颈癌患者（宫颈癌组）、80例CIN患者（CIN组）和60例健康体检正常女性（对照组）,检测各组血清miR-196a、SCC-Ag及CEA水平,分析其与宫颈癌患者临床病理特征的关系。应用ROC曲线分析血清miR-196a、SCC-Ag及CEA水平对宫颈癌的诊断价值。Pearson相关分析血清miR-196a与SCC-Ag及CEA水平的相关性。结果: 宫颈癌组血清miR-196a（5.26±1.25 vs 1.65±0.41,1.18±0.24）、SCC-Ag（ng/ml）（4.16±1.20 vs 0.68±0.25,0.52±0.13）及CEA（ng/ml）（14.50±4.17 vs 3.72±0.81,3.05±0.63）水平均明显高于CIN组和对照组（P<0.01）。宫颈癌患者血清miR-196a表达水平与临床分期、淋巴结转移、浸润深度、SCC-Ag及CEA阳性有关（P<0.05）。ROC曲线分析显示,血清miR-196a、SCC-Ag及CEA水平诊断宫颈癌的最佳截断值分别为3.84、2.37 ng/ml、9.60 ng/ml,三项联合诊断宫颈癌的曲线下面积最大（0.912,95%CI:0.854～0.974）,其敏感度和特异度为90.4%和86.5%。相关分析显示,宫颈癌患者血清miR-196a与SCC-Ag及CEA水平均呈正相关（r=0.817,P<0.01,r=0.748,P<0.01）。结论: 血清miR-196a表达水平在宫颈癌患者中明显升高,且与SCC-Ag及CEA水平均呈正相关,三项联合检测对宫颈癌诊断具有较高的价值。     

结直肠癌中医辨证分型与miR-29、miR-34a表达的相关性分析

目的:探究结直肠癌患者血清miR-29、miR-34a表达水平及其与中医辨证分型的相关性。方法:选取2014年06月至2015年06月本院收治的127例结直肠癌患者作为研究对象（结直肠癌组）,并根据中医辨证分型标准分为湿热内蕴型21例、气滞血瘀型40例、脾肾阳虚型29例、气血两虚型18例、肝肾阴虚型19例;另选取同期在本院体检的127例健康者作为健康对照组。采用实时荧光定量PCR（qRT-PCR）技术检测血清miR-29、miR-34a表达水平,并进行比较;采用Pearson法分析血清miR-29与miR-34a表达水平的相关性;采用Logistic回归模型分析影响结直肠癌患者不良预后发生的危险因素。结果:结直肠癌组患者血清miR-29、miR-34a表达水平均明显低于健康对照组（P＜0.05）。低分化、浸润程度T3-T4、TNM分期Ⅲ-Ⅳ期、淋巴结转移、远处转移患者血清miR-29、miR-34a表达水平明显低于高中分化、浸润程度T1-T2、TNM分期Ⅰ-Ⅱ期、无淋巴结转移、无远处转移患者（P＜0.05）。结直肠癌患者血清miR-29与miR-34a表达水平呈正相关（r=0.529,P=0.000）。低分化、浸润程度T3-T4、TNM分期Ⅲ-Ⅳ期、淋巴结转移、低miR-29水平、低miR-34a水平是影响结直肠癌患者不良预后发生的危险因素（P＜0.05）。脾肾阳虚型、气滞血瘀型、湿热内蕴型预后不良患者比例依次明显增加（P＜0.05）。肝肾阴虚型、气血两虚型、脾肾阳虚型、气滞血瘀型、湿热内蕴型结直肠癌患者血清miR-34a表达水平依次显著降低（P＜0.05）,miR-29表达水平差异无统计学意义（P＞0.05）。结论:结直肠癌患者血清中miR-29、miR-34a低表达,与肿瘤分期升高、分化程度降低、浸润程度增加、淋巴结转移、远处转移、不良预后等密切相关,且miR-34a对结直肠癌不同中医辨证分型有一定的分型参考意义。     

上皮性卵巢癌血清miR-1294及miR-4443的表达及其与预后的关系

目的:探讨上皮性卵巢癌（epithelial ovarian cancer,EOC）患者血清miR-1294及miR-4443的表达水平及其与患者临床病理特征和预后的关系。方法:选取我院2015年1月至2017年12月收治的106例EOC患者作为EOC组,另选取90例卵巢良性肿瘤患者作为良性组和60例正常健康女性作为对照组,检测血清miR-1294及miR-4443表达水平。以miR-1294及miR-4443的最佳截断值为标准将EOC患者分为高miR-1294组（n=33）、低miR-1294组（n=73）和高miR-4443组（n=36）、低miR-4443组（n=70）。EOC患者预后不良的危险因素应用单因素及多因素COX回归模型进行分析。结果:血清miR-1294表达水平（0.48±0.13 vs 1.16±0.57、1.21±0.62）在EOC组明显低于良性组和对照组（P均<0.001）。血清miR-4443表达水平（0.71±0.18 vs 1.36±0.64、1.45±0.70）在EOC组明显低于良性组和对照组（P均<0.001）。miR-1294和miR-4443高表达组的EOC患者临床分期、分化程度、淋巴结转移及腹膜转移与miR-1294和miR-4443低表达组比较,差异均有统计学意义（P<0.05）。Kaplan-Meier分析显示,EOC患者生存期短与miR-1294及miR-4443低表达有关（P<0.001）。多因素分析显示,EOC患者预后不良的独立危险因素为淋巴结转移［HR（95%CI）=1.885（1.206～4.118）］、腹膜转移［HR（95%CI）=3.106（2.315～6.226）］、miR-1294＜0.73［HR（95%CI）=2.614（1.865～5.512）］及miR-4443＜1.04［HR（95%CI）=1.975（1.508～4.903）］。结论:miR-1294及miR-4443的低表达与EOC患者生存期短有关,是EOC患者预后预测的生物标志物。     

miR-196a调控HOXA5对人胃癌细胞MGC-803侵袭转移的影响及机制研究

目的 检测miR-196a在人胃癌组织及细胞系中的表达,探讨抑制或过表达miR-196a对胃癌细胞侵袭转移能力的影响,以及其可能作用的靶基因。方法 通过实时定量PCR技术检测胃癌组织及细胞系中miR-196a的表达水平,通过转染miR-196a inhibitors或mimics抑制或上调其表达,并通过定量PCR检测转染效率。利用划痕迁移实验、Transwell侵袭实验和MTT实验检测上调或下调miR-196a水平对MGC-803细胞的迁移、侵袭和增殖能力的影响。采用生物信息学及Western blotting方法验证miR-196a对靶基因HOXA5的调控机制。结果 相对于正常胃黏膜组织及细胞,胃癌组织和细胞系中miR-196a的表达水平显著上调（上调约28倍,P＜0.01）,MGC-803细胞中转染miR-196a inhibitors或mimics能显著抑制（下降了53％,P＜0.01）或上调（上调约8倍,P＜0.01）miR-196a表达水平。抑制miR-196a表达能降低MGC-803细胞的迁移、侵袭和增殖能力,而上调其表达则相反。miR-196a能够负性调控HOXA5的表达。结论 胃癌组织及细胞系中miR-196a的表达上调可能通过抑制HOXA5的表达显著提高胃癌细胞的侵袭转移能力,促进胃癌的发生、发展。     

肝细胞癌患者外周血中miR-30a、miR-106b的表达及其与预后的关系

目的:探讨肝细胞癌(hepatocellular carcinoma,HCC)患者血清miR-30a、miR-106b的表达及其与 预后的关系。方法:选取2015年6月-2017年2月本院收治的HCC患者80例为研究对象,20例同期健康人群作为对照组。荧光定量PCR检测外周血中miR-30a、miR-106b的表达水平,分析患者血清miR-30a、miR-106b水平与HCC临床病理的关系。分析HCC患者预后的影响因素以及血清miR-30a、miR-106b水平与HCC患者预后的相关性。结果:HCC患者血清miR-30a表达水平（1.03±0.02）低于正常人群（5.15±0.06）（P=0.00）;血清miR-106b表达水平（2.62±0.35）高于正常人群（1.15±0.06）（P=0.00）。miR-30a、miR-106b表达与患者肿瘤直径、组织分化类程度、肝内转移、淋巴结转移、甲胎蛋白（AFP）水平、门静脉癌栓、肿瘤数目及TNM分期有显著相关性(P＜0.05) 。miR-30a低表达组患者3年总生存率（39.8%）低于miR-30a高表达组（64.6%）（P=0.002）;miR-106b低表达组患者3年总生存率（75.8%）高于miR-106b高表达组（51.2%）（P=0.003）。二元Logistic回归分析提示,淋巴结转移、TNM分期、miR-106b及miR-30a水平是患者预后的重要影响因素。结论:miR-30a在HCC患者血清中低表达,miR-106b在HCC患者血清中高表达,与肿瘤的进展、预后不良有关。     

Identification of lncRNA TRPM2-AS/miR-140-3p/PYCR1 axis’s proliferates and anti-apoptotic effect on breast cancer using co-expression network analysis

Breast cancer (BC) is one of the most common malignancies occurring in women worldwide. Weighted gene co-expression network analysis (WGCNA) has not been widely utilized in uncovering the biomarkers which played pivotal roles in BC treatment. This study aimed to verify the proliferative and anti-apoptotic effect of lncRNA TRPM2-AS/miR-140-3p/PYCR1 axis on BC based on WGCNA. WGCNA was applied for determining hub genes using gene expression data gained from breast cancer and adjacent tissues which were downloaded from the Cancer Genome Atlas (TCGA) database. The correlative curves showed the correlation between OS/DFS of BC patients and TRPM2-AS expression or PYCR1 expression based on the data of survival rate of BC patients obtained from the TCGA database. QRT-PCR was employed in detecting the expression levels of TRPM2-AS, miR-140-3p and PYCR1, and western blot analysis was adopted for determination of protein expression level of PYCR1. Dual luciferase assay was applied to verify the targeting relationship between TRPM2-AS and miR-140-3p, as well as miR-140-3p and PYCR1. The roles of TRPM2-AS, miR-140-3p, and PYCR1 in proliferation, migration, and apoptosis of BC cell were identified by CCK-8 assay, cell migration assay and flow cytometry. Hub genes were also gained from WGCNA test. The prognostic study showed a significant negative correlation between the high expression of PYCR1 and TRPM2-AS and the BC survival. QRT-PCR demonstrated that PYCR1 and TRPM2-AS were both overexpressed, while miR-140-3p was greatly down-regulated in BC cell. In addition, it was validated by dual luciferase assay that miR-140-3p directly targeted both TRPM2-AS and PYCR1. Furthermore, down-regulation of TRPM2-AS and PYCR1 inhibited proliferation yet promoted apoptosis of BC cell, and up-regulation of miR-140-3p in BC cell showed the same tendency. Taken together, TRPM2-AS could promote proliferation and inhibit apoptosis of BC cell through TRPM2-AS/miR-140-3p/PYCR1 axis.