Validation of a serum microRNA panel as biomarkers for early diagnosis of hepatocellular carcinoma post-hepatitis C infection in Egyptian patients

AIM To investigate the prospective importance of serum micro(mi)RNAs(mi R-125 b, mi R-138 b, mi R-1269, mi R-214-5p, mi R-494, mi R375 and mi R-145) as early biomarkers for the diagnosis of hepatitis C virus(HCV)-related hepatocellular carcinoma(HCC).METHODS Two-hundred and fifty HCV4 a patients, 224 HCV4 aHCC patients, and 84 healthy controls were enrolled in the study. Expression levels of mi R214-5p, mi R-125 b, mi R-1269 and mi R-375 were quantified using quantitative real-time PCR.RESULTS Expression of the selected mi RNAs in serum wassignificantly lower in HCC patients than in the healthy controls, except for mi R-1269 and mi R-494. There was a significant difference between HCC and HCV patients, in particular for HCC and late stage fibrosis, rather than HCV patients and early fibrosis. It is obvious that mi R-1269 was significantly upregulated in HCC cases compared to hepatic fibrosis cases. Each mi RNA can show HCC progression. Multivariate logistic regression analysis indicated that the tested panel of mi RNAs(mi R214-5p, mi R-125 b, mi R-1269 and mi R-375) represent accurate and specific indictors of HCC development.CONCLUSION This study presents a panel of mi RNAs with strong power as putative diagnostic and prognostic biomarkers for HCV-induced HCC. Moreover, mi R-214-5p and mi R-1269 could be considered as early biomarkers for tracking the progress of liver fibrosis to HCC.

Autophagy and microRNA in hepatitis B virus-relatedhepatocellular carcinoma

Approximately 350 million people worldwide are chronically infected by hepatitis B virus(HBV).HBV causes severe liver diseases including cirrhosis and hepatocellular carcinoma(HCC).In about 25%of affected patients,HBV infection proceeds to HCC.Therefore,the mechanisms by which HBV affects the host cell to promote viral replication and its pathogenesis have been the subject of intensive research efforts.Emerging evidence indicates that both autophagy and micro RNAs(mi RNAs)are involved in HBV replication and HBV-related hepatocarcinogenesis.In this review,we summarize how HBV induces autophagy,the role of autophagy in HBV infection,and HBV-related tumorigenesis.We further discuss the emerging roles of mi RNAs in HBV infection and how HBV affects mi RNAs biogenesis.The accumulating knowledge pertaining to autophagy and mi RNAs in HBV replication and its pathogenesis may lead to the development of novel strategies against HBV infection and HBV-related HCC tumorigenesis.

Studied microRNA gene expression in human hepatocellular carcinoma by micro RNA microarray techniques

AIM: To achieve a better understanding of the molecular mechanisms of micro RNA expression changes involved in hepatocellular carcinoma.METHODS: In this research process, patients were not treated with antivirals, immunosuppressants or immunomodulators for at least 6 mo before collecting serum. The study population was composed of 35 outpatient hepatitis B virus(HBV) cases and 12 healthy control cases from the Affiliated Hospital of Inner Mongolia Medical University(Inner Mongolia, China) from July 2013 to April 2014. The 35 HBV cases were divided into two groups: a hepatocirrhosis group with 20 cases and a liver cancer group with 15 cases. All 35 cases carried HBs Ag. The diagnostic criteria followed the European Association for the Study of the Liver 2012(EASL2012) standards. Micro RNA(mi RNA) was extracted from a control group of patients, a group with hepatocirrhosis and a group with liver cancer and its quality was analyzed using the human V2 micro RNA expression beadchip. Cluster analysis and a radar chart were then applied to the mi RNA changes.RESULTS: The mi RNA-qualified rate of human serum samples was 93%. The concentration of a single sample was > 200 ng/μL and the volume was > 5 μL.All mi RNA serum samples were uncontaminated by the genome. The Mann-Whitney test showed significant differences in mi RNA between each group, with a detection P-value of < 0.05. Illumina software was set up with Diff Score set to ± 13, meaning that P = 0.001.There were significant changes in mi RNA expression between the three groups. mi RNA-183 was the most up-regulated, followed by mi RNA-373. mi RNA-129 and mi RNA-188 were both strongly down-regulated and mi RNA-378 was down-regulated a small amount. The liver cancer group had greater changes, which indicated that changes in mi RNA expression levels were caused by hepatocirrhosis. The liver cancer disease course then further increased these changes. In the pentagon created by these five mi RNAs, three groups showed significant deviation. The liver cancer group had a bigger deviation trend. The chart indicated that mi RNA expression changes occurred in the hepatocirrhosis group, which increased in the liver cancer disease course and were irreversible.CONCLUSION: There was a significant relationship between the irreversible up-regulation of mi RNA-183/373 and down-regulation of mi RNA-129/188/378 and incidences of hepatocirrhosis and liver cancer.

Up-regulation of micro RNA-210 inhibits proliferation of hepatocellular carcinoma cells by targeting YES1

AIM: To determine the expression of micro RNA-210(mi R-210) in hepatocellular carcinoma(HCC) and to examine its role using HCC cells.METHODS: The expression of mi R-210 was determined in 21 pairs of HCC samples and the corresponding surrounding non-tumor tissues. The effects of mi R-210 on proliferation and cell cycle progression were examined using Hep G2 and Hu H7 cells. Overexpression and inhibition of mi R-210 was achieved by transfection of the cells with mi R-210 mimic or inhibitor. Luciferase reporter constructs were used to identify the mi R-210 interacting site on Yes1. Yes1 expression was examined after mi R-210 transfection,as well as in the HCC samples.RESULTS: mi R-210 was significantly up-regulated by 3.4 fold(P < 0.01) in the tumor samples. The over-expression of mi R-210 significantly reduced cell proliferation compared to the mock-treated cells(68.9% ± 7.4% and 53.6% ± 5.0%,P < 0.05 for the Hep G2 and Hu H7 cells respectively). Analysis of the Hu H7 cells transfected with mi R-210 mimic by flow cytometry showed that the cells took a longer time to reach the G2/M phase. The interaction between mi R-210 and the 3'UTR of the Yes1 transcript was confirmed using a luciferase reporter assay. Over-expression of mi R-210 reduced the expression of Yes1 protein in both Hu H7 and Hep G2 cells. Tumors with a greater than fourfold increase in the expression of mi R-210 showed consistently lower expressions of Yes1 in the tumors.In nocodazole-treated cells with a significant G2/M cell population,Yes1 protein was significantly reduced and pre-inhibition of mi R-210 in Hu H7 cells was able to prevent the reduction of Yes1 protein expression. Knock-down of Yes1 by si RNA also led to reduced cell proliferation(70.8% ± 7.5%,P < 0.05 in the Hu H7 cells).CONCLUSION: Up-regulation of mi R-210 inhibits cell proliferation. Yes1 is a target of mi R-210 and affects cell proliferation in HCC.

MicroRNA signature in patients with hepatocellular carcinoma associated with type 2 diabetes

BACKGROUND Nonalcoholic steatohepatitis-related cirrhosis is one of the liver complications in type 2 diabetes mellitus(T2DM)and reported to be a risk factor for developing hepatocellular carcinoma(HCC).A reliable screening biomarker of liver cirrhosis(LC)and HCC among T2DM patients is important to reduce the morbidity and mortality of this disease.MicroRNA(miRNA)is considered a key player in HCC and T2DM,and it might be a hidden culprit in diabetes-associated HCC,making it a promising reliable prognostic tool.AIM To investigate the signature of serum miRNAs as early biomarkers for the screening of HCC among diabetic patients.METHODS Expression profiles of miRNAs in serum samples of diabetic LC and diabetic HCC patients were assessed using Illumina sequencing;then,RT-qPCR was used to validate significantly altered miRNAs between the two groups.Candidate miRNAs were tested in serum samples of 200 T2DM patients,270 LC patients,200 HCC patients,and 225 healthy control subjects.Additionally,receiver operating characteristic(ROC)analysis,with area under the curve(AUC),was performed to assess the diagnostic performance of the screened miRNAs for discriminating HCC from LC and nonmalignant patients(LC+T2DM).RESULTS Expression of the sequenced miRNAs in serum was different in HCC vs LCpositive T2DM patients.Two miRNAs(miR-34a,miR-221)were significantly upregulated and five miRNAs(miR-16,miR-23-3p,miR-122-5p,miR-198,miR-199a-3p)were significantly down-regulated in HCC compared to LC patients.Analysis of ROC curve demonstrated that the combination of these seven miRNAs can be used as a reliable biomarker for detection of HCC in diabetic patients,as it could identify HCC with high diagnostic accuracy in diabetic LC patients(AUC=0.993)and in diabetic nonmalignant patients(AUC=0.961).CONCLUSION This study validates a panel of serum miRNAs that can be used as a reliable noninvasive screening biomarker of HCC among T2DM cirrhotic and noncirrhotic patients.The study recommends further research to shed light on a possible role of c-Met in T2DM-associated HCC via the miRNA regulatory pathway.

MicroRNAs dysregulation in hepatocellular carcinoma: Insights in genomic medicine

Hepatocellular carcinoma(HCC) is the leading primary liver cancer and its clinical outcome is still poor. MicroRNAs(mi RNAs) have demonstrated an interesting potential to regulate gene expression at post-transcriptional level. Current findings suggest that mi RNAs deregulation in cancer is caused by genetic and/or epigenetic, transcriptional and post-transcriptional modifications resulting in abnormal expression and hallmarks of malignant transformation: aberrant cell growth, cell death, differentiation, angiogenesis, invasion and metástasis. The important role of mi RNAs in the development and progression of HCC has increased the efforts to understand and develop mechanisms of control overt this single-stranded RNAs. Several studies have analyzed tumoral response to the regulation and control of deregulated mi RNAs with good results in vitro and in vivo, proving that targeting aberrant expression of mi RNAs is a powerful anticancer therapeutic. Identification of up and/or down regulated mi RNAs related to HCC has led to the discovery of new potential application for detection of their presence in the affected organism. Mi RNAs represent a relevant new target for diagnosis, prognosis and treatment in a wide variety of pathologic entities, including HCC. This manuscript intends to summarize current knowledge regarding mi RNAs and their role in HCC development.

Therapeutic potential of small interfering RNAs/micro interfering RNA in hepatocellular carcinoma

Hepatocellular carcinoma(HCC) is the predominant form of primary liver cancer and represents the third leading cause of cancer-related death worldwide. Current available therapeutic approaches are poorly effective,especially for the advanced forms of the disease. In the last year,short double stranded RNA molecules termed small interfering RNAs(si RNAs) and micro interfering RNAs(mi RNA),emerged as interesting molecules with potential therapeutic value for HCC. The practical use of these molecules is however limited by the identification of optimal molecular targets and especially by the lack of effective and targeted HCC delivery systems. Here we focus our discussion on the most recent advances in the identification of si RNAs/mi RNAs molecular targets and on the development of suitable si RNA/mi RNAs delivery systems.

Current biomarkers for hepatocellular carcinoma: Surveillance, diagnosis and prediction of prognosis

Biomarkers for surveillance, diagnosis and prediction of prognosis in patients with hepatocellular carcinoma(HCC) are currently not ready for introduction into clinical practice because of limited sensitivity and specificity. Especially for the early detection of small HCC novel biomarkers are needed to improve the current effectiveness of screening performed byultrasound. The use of high-throughput technologies in hepatocellular research allows to identify molecules involved in the complex pathways in hepatocarcinogenesis. Several invasive and non-invasive biomarkers have been identified already and have been evaluated in different clinical settings. Gene signatures with prognostic potential have been identified by gene expression profiling from tumor tissue. However, a single "all-in-one" biomarker that fits all-surveillance, diagnosis, prediction of prognosis-has not been found so far. The future of biomarkers most probably lies in a combination of non-invasive biomarkers, imaging and clinical parameters in a surveillance setting. Molecular profiling of tumorous and non-tumorous liver tissue may allow a prediction of prognosis for the individual patient and hopefully clear the way for individual treatment approaches. This article gives an overview on current developments in biomarker research in HCC with a focus on currently available and novel biomarkers, in particular on micro RNA.

Liver expression of steroid hormones and Apolipoprotein D receptors in hepatocellular carcinoma

AIM: To evaluate the tissular expression of Androgen (A), Estrogen (E) and Progesterone (Pg) receptors, and Apolipoprotein D (ApoD), in liver tumors from resected hepatocellular carcinoma (HCC) cases in order to assess their possible relationship to prognosis. METHODS: We performed an immunohistochemical study using tissue microarrays (containing more than 260 cancer specimens, from 31 HCC patients and controls) to determine the presence of specif ic antibodies against AR, ER, PgR and ApoD, correlating their findings with several clinico-pathological and biological variables. The staining results were categorized using a semi-quantitive score based on their intensity, and the percentage of immunostained cells was measured. RESULTS: A total of 21 liver tumors (67.7%) were positive for AR; 16 (51.6%) for ER; 26 (83.9%) for PgR and 12 (38.7%) stained for ApoD. We have found a wide variability in the immunostaining score values for each protein, with a median (range) of 11.5 (11.5-229.5) for AR; 11.1 (8.5-65) for ER; 14.2 (4-61) for PgR; and 37.7 (13.8-81.1) for ApoD. A history of heavy ethanol consumption, correlated positively with AR and PgR and negatively with ER status. HCV chronic infection also correlated positively with AR and PgR status. However, the presence of ApoD immunostaining did not correlate with any of these variables. Tumors with a positive immuno-staining for PgR showed a better prognosis. CONCLUSION: Our results indicate a moderate clinical value of the steroid receptor status in HCC, emphasizing the need to perform further studies in order to evaluate the possible role of new hormonal-based therapies.

Diagnostic and therapeutic application of noncoding RNAs for hepatocellular carcinoma

Micro RNAs(mi RNAs) are small,noncoding RNA molecules that regulate gene expression posttranscriptionally,targeting thousands of messenger RNAs. Long noncoding RNAs(lnc RNAs),another class of noncoding RNAs,have been determined to be also involved in transcription regulation and translation of target genes. Since deregulated expression levels or functions of miR NAs and lncR NAs in hepatocellular carcinoma(HCC) are frequently observed,clinical use of noncoding RNAs for novel diagnostic and therapeutic applications in the management of HCCs is highly and emergently e xpe c t e d. H e r e,we s ummar iz e r e c e nt f indings regarding deregulated mi RNAs and lnc RNAs for their potential clinical use as diagnostic and prognostic biomarkers of HCC. Specifically,we emphasize the deregulated expression levels of such noncoding RNAs in patients' sera as noninvasive biomarkers,a field that requires urgent improvement in the clinical surveillance of HCC. Since nucleotide-based strategies are being applied to clinical therapeutics,we further summarize clinical and preclinical trials using oligonucleotides involving the use of miR NAs and small interfering RNAs against HCC as novel therapeutics. Finally,we discuss current open questions,which must be clarified in the near future for realistic clinical applications of these new strategies.

Novel hepatocellular carcinoma molecules with prognostic and therapeutic potentials

Hepatocellular carcinoma(HCC), the predominant form of primary liver cancer, is the sixth most common cancer worldwide and the third leading cause of cancerrelated death. The difficulty to diagnose early cancer stages, the aggressive behaviors of HCC, and the poor effectiveness of therapeutic treatments, represent the reasons for the quite similar deaths per year and incidence number. Considering the fact that the diagnosis of HCC typically occurs in the advanced stages of the disease when the therapeutic options have only modest efficacy, the possibility to identify early diagnostic markers could be of significant benefit. So far, a large number of biomarkers have been associated to HCC progression and aggressiveness, but many of them turned out not to be of practical utility. This is the reason why active investigations are ongoing in this field. Given the huge amount of published works aimed at the identification of HCC biomarkers, in this review we mainly focused on the data published in the last year, with particular attention to the role of(1) molecular and biochemical cellular markers;(2) micro-interfering RNAs;(3) epigenetic variations; and(4) tumor stroma. It is worth mentioning that a significant number of the HCC markers described in the present review may be utilized also as targets for novel therapeutic approaches, indicating the tight relation between diagnosis and therapy. In conclusion, we believe that integrated researches among the different lines of investigation indicated above should represent the winning strategies to identify effective HCC markers and therapeutic targets.

MicroRNA-33b通过靶向调控SALL4的表达抑制肝细胞癌细胞增殖

目的:探索微小RNA-33b(micro RNA-33b,mi R-33b)在肝细胞癌(hepatocellular carcinoma,HCC)中的表达及调控HCC细胞增殖的相关分子机制。方法:收集配对的HCC及癌旁组织32对,分别采用实时荧光定量PCR和Western印迹法检测人类婆罗双树样基因4(Sal-like 4,SALL4)RNA和蛋白表达,MTT实验检测HCC细胞增殖,萤光素酶报告基因检测验证mi R-33b与SALL4基因的靶点关系。结果:Mi R-33b在HCC组织中的表达显著低于癌旁组织(P<0.001)。过表达mi R-33b能显著降低HCC LH86细胞的增殖。SALL4是mi R-33b的靶基因,其蛋白表达被mi R-33b负调控。过表达SALL4能逆转mi R-33b对LH86细胞增殖的抑制作用。SALL4在HCC组织中的表达显著高于癌旁组织(P<0.001)。结论:Mi R-33b对HCC细胞增殖的抑制作用是通过负调控SALL4的表达而实现。

MicroRNAs在肝细胞肝癌中作用的研究进展

MicroRNAs(miRNAs)是近年来发现的长度约为22 nt的一类非编码小分子RNA,通过在转录后或翻译水平调节靶基因功能,进而在生物个体发育、细胞增殖、凋亡、分化、肿瘤发生等生理及病理过程中发挥重要作用.miRNAs可能通过参与调控癌基因和抑癌基因的表达,促进恶性肿瘤的发生发展,或者其本身就是潜在的癌基因或抑癌基因.越来越多的研究显示miRNAs与肝细胞肝癌(hepatocellular carcinoma,HCC)关系密切.本文就miRNAs在HCC发生发展中的作用以及在HCC诊断与治疗中的应用价值等方面的研究进展作一综述.

Micro-PET在膈下逐瘀汤抑制肝细胞癌研究中的应用

目的采用micro-PET显像技术评判中药复方膈下逐淤汤对荷肝细胞癌裸鼠的抑制作用。方法 20周龄健康裸鼠12只,随机分为给药组与对照组(n=6),皮下接种人肝癌细胞Bel7402建立荷瘤模型。接种1周后每日灌胃膈下逐淤汤0.3 ml,对照组给予等体积生理盐水,连续给药60 d。待肿瘤长至约5 mm时测量肿瘤体积,每周2次,计算瘤体积大小和抑瘤率并绘制肿瘤生长曲线。给药24、60 d后分别行^(18)F-FDG、^(18)F-RGD micro-PET扫描各一次,测定肿瘤组织摄取量、标准摄取值(SUV)及肿瘤/肌肉靶本比(T/NT)。结果接种24 d后,肿瘤生长至5 mm,给药组瘤体积(74.8 mm^3)小于对照组(78.3mm^3)。^(18)F-FDG micro-PET扫描给药组的肿瘤组织摄取量、SUV_(mean)和T/NT分别为(5.54±1.59)%ID/g、0.93±0.20和8.20±2.52,略小于对照组(5.92±1.23)%ID/g、1.00±0.19和8.71±2.36(P>0.05)。^(18)F-RGD micro-PET扫描给药组的肿瘤组织摄取量、SUV和T/NT分别为(4.08±0.64)%ID/g、0.75±0.08和6.91±0.72,略小于对照组(4.61±1.08)%ID/g、0.87±0.16和7.00±1.40(P>0.05)。接种60 d后,给药组和对照组的肿瘤体积分别增长至1854.4 mm^3和1462.9 mm^3。^(18)F-FDG micro-PET扫描给药组的肿瘤组织摄取量、SUV和T/NT分别为(3.56±0.54)%ID/g、0.70±0.09和4.91±0.92(n=5),大于对照组的3.28%ID/g、0.60和3.98(n=1)。^(18)F-RGD micro-PET扫描给药组的肿瘤组织摄取量、SUV和T/NT分别为(2.19±0.16)%ID/g、0.51±0.04和4.15±0.57(n=4)。给药组和对照组的肿瘤重量分别为(1.93±0.95)g(n=5)和1.69 g(n=1),抑瘤率为-14.76%。给药组的中位生存时间为60 d,长于对照组的40.5 d,差异有统计学意义(P<0.05)。结论膈下逐淤汤可能通过抑制葡萄糖的吸收和新生血管的生成来延长荷肝细胞癌裸鼠的生存时间。Micro-PET能动态地定量分析活体动物模型的生理、生化变化,可用于评价抗肿瘤药物的体内疗效。

负载人microRNA-149的复制缺陷型腺病毒对人肝癌细胞株增殖的影响

目的包装负载人micro RNA-149的复制缺陷型腺病毒(Ad-mi R149),并检测其对人肝癌细胞株增殖的影响。方法全基因合成micro RNA-149前体核苷酸序列,插入腺病毒穿梭质粒的多克隆位点,随后与包装质粒共转染人胚肾293细胞,包装负载micro RNA-149的复制缺陷型腺病毒;采用MMT方法检测Ad-mi R149对人肝癌细胞株增殖的影响。结果成功包装负载人micro RNA-149的复制缺陷型腺病毒,采用倍比稀释法检测病毒滴度为5×109 pfu/ml。采用电子显微镜技术,观察到HEK293细胞中大量包装病毒颗粒。采用PCR扩增,Ad-mi R149可获得腺病毒及micro RNA-149特异片段,而对照AdLac Z只能扩增出腺病毒特异片段;采用MTT方法发现,Ad-mi R149可显著抑制肝癌细胞株增殖。结论成功包装负载人micro RNA-149的复制缺陷型腺病毒并初步证实其对肝癌细胞增殖的抑制作用。

Hepatitis B virus and micro RNAs:Complex interactions affecting hepatitis B virus replication and hepatitis B virusassociated diseases

Chronic infection with the hepatitis B virus(HBV) is the leading risk factor for the development of hepatocellular carcinoma(HCC). With nearly 750000 deaths yearly, hepatocellular carcinoma is the second highest cause of cancer-related death in the world. Unfortunately, the molecular mechanisms that contribute to the development of HBV-associated HCC remain incompletely understood. Recently, micro RNAs(mi RNAs), a family of small non-coding RNAs that play a role primarily in post-transcriptional gene regulation, have been recognized as important regulators of cellular homeostasis, and altered regulation of mi RNA expression has been suggested to play a significant role in virus-associated diseases and the development of many cancers. With this in mind, many groups have begun to investigate the relationship between mi RNAs and HBV replication and HBV-associated disease. Multiple findings suggest that some mi RNAs, such as mi R-122, and mi R-125 and mi R-199 family members, are playing a role in HBV replication and HBV-associated disease, including the development of HBV-associated HCC. In this review, we discuss the current state of our understanding of the relationship between HBV and mi RNAs, including how HBV affects cellular mi RNAs, how these mi RNAs impact HBV replication, and the relationship between HBV-mediated mi RNA regulation and HCC development. We also address the impact of challenges in studying HBV, such as the lack of an effective model system for infectivity and a reliance on transformed cell lines, on our understanding of the relationship between HBV and mi RNAs, and proposepotential applications of mi RNA-related techniques that could enhance our understanding of the role mi RNAs play in HBV replication and HBV-associated disease, ultimately leading to new therapeutic options and improved patient outcomes.

Micro RNA expression in hepatitis C virus-related malignancies: A brief review

Not only is chronic hepatitis C virus(HCV) infection a major public health problem,but also it can cause hepatocellular carcinoma and,more rarely,nonHodgkin's lymphoma.These characteristics mean that HCV is the only virus infecting humans that is able to cause two different cancers.The fine pathogenetic and molecular mechanisms by which HCV induces these two malignancies are not completely clear.In the last decade,it has been shown that microRNAs(miRNAs),a class of 21-23-nucleotide molecules modulating posttranscriptional gene expression,make an important contribution to the pathogenesis of several cancers and are also considered highly promising biomarkers.Here,we briefly describe the current knowledge about microRNAs' involvement in HCV-related molecular oncogenesis.We decided to focus our attention on studies fully conducted on ex vivo samples with this specific etiology,and on cultured cell lines partially or completely expressing the HCV genome.Some of the results reported in this review are controversial,possibly because of methodological issues,differences in sampling size and features,and ethnicity of patients.What is certain is that miRNAs play a remarkable role in regulating gene expression during oncogenetic processes and in viral infection.A clear understanding of their effects is fundamental to elucidating the mechanisms underlying virus-induced malignancies.

microRNA-451在肝癌患者血浆中的表达及意义

目的探讨micro RNA-451(mi R-451)在肝癌患者血浆中的表达水平及其临床意义。方法选择2015年3~10月在枣庄矿业集团中心医院住院治疗的42例肝癌患者和42例健康体检者作为研究对象,利用定量逆转录聚合酶链反应(RT-PCR)方法检测血浆中mi R-451的表达水平,分析肝癌患者血浆中mi R-451表达水平与临床病理参数的关系。结果 mi R-451在肝癌患者血浆中表达水平(0.43±0.16)明显低于对照组(0.59±0.21),差异有高度统计学意义(P<0.01);肝癌患者血浆中mi R-451水平在不同性别、年龄、分化程度及甲胎蛋白水平间差异无统计学意义(P>0.05),而与肿瘤大小、淋巴转移、静脉侵犯、肿瘤数量及TNM分期明显相关(P<0.05)。结论肝癌患者血浆中mi R-451表达水平降低,可能参与了肝癌的发病过程。

CEUS动脉期诊断肝硬化背景下微小肝细胞癌与病理结果的一致性分析

目的 研究超声造影(CEUS)动脉期诊断肝硬化背景下微小肝细胞癌的价值。方法 纳入2016年8月—2021年8月长沙市第三医院收治的肝结节患者66例(共70个结节)为研究对象,所有患者均存在肝硬化背景。患者均经病理检查明确病变性质,并据此分为微小肝癌组(38例、40个结节)和非恶变结节组(28例、30个结节)。病理诊断前,患者均行CEUS检查,观察各病灶CEUS增强灌注模式及增强峰值、降支减半及持续增强时间,分析CEUS诊断肝硬化背景下微小肝细胞癌与病理结果的一致性。结果 微小肝癌组所有患者动脉相造影均呈现高增强,而门脉相则多为低增强(32/40,80.00%);非恶变组病灶动脉相以低增强为主(17/30,56.67%),门脉相以等增强为主(22/30,73.33%)。两组病灶在动脉相和门脉相的增强水平有显著差异(P<0.05)。微小肝癌组降支减半时间(84.68±18.20)s、增强峰值时间(22.65±8.84)s、持续增强时间(41.03±10.81)s均显著低于非恶变组[(115.03±20.12)s、(30.59±10.75)s、(84.54±18.20)s](P<0.05)。CEUS诊断肝硬化后微小肝细胞癌的灵敏度为92.50%,特异度为80.00%,准确率为87.14%,阳性预测值为86.05%,阴性预测值为88.89%,Kappa值为0.734。结论 肝硬化背景下微小肝细胞癌与肝硬化结节有不同的CEUS灌注特点,CEUS对早期诊断肝硬化合并微小肝癌具有重要价值。

原发性肝癌患者血清miR-196b和miR-520f表达水平及其临床诊断价值分析

目的 探讨原发性肝癌(primary carcinoma of the liver)患者血清微小核糖核酸(microRNA,miR)-196b,微小核糖核酸(microRNA,miR)-520f表达水平及其临床诊断价值。方法 选择2020年6月~2022年6月攀枝花学院附属医院收治的73例原发性肝癌患者作为研究组,患者均符合《原发性肝癌诊疗规范》中的诊断标准;选择同期医院体检的80例健康人群作为对照组。抽取研究对象清晨空腹外周静脉血5ml,采用实时荧光定量逆转录-聚合酶链反应(real time fluorescent quantitative reverse transcription-polymerase chain reaction,qRT-PCR)检测两组血清miR-196b和miR-520f表达水平。分析不同临床病理特征原发性肝癌患者血清miR-196b和miR-520f表达水平差异,采用Pearson相关性分析探讨血清miR-196b与miR-520f的关系,采用受试者工作特征(receiver operating characteristic,ROC)曲线分析血清miR-196b和miR-520f对原发性肝癌的诊断价值。结果 研究组血清miR-196b表达水平(2.73±0.56)明显高于对照组(0.99±0.24),miR-520f表达水平(1.69±0.44)明显低于对照组(3.34±0.64),差异具有统学意义(t=30.578,12.690,均P<0.05)。与Ⅰ~Ⅱ期、中高分化、无淋巴结转移、肿瘤直径<5cm患者相比,Ⅲ~Ⅳ期、低分化、淋巴结转移和肿瘤直径≥5cm患者血清miR-196b表达水平均升高,miR-520f表达水平均降低,差异具有统计学意义(tmiR-196b=6.919~13.229,tmiR-520f=3.873~7.814,均P<0.05)。Pearson相关性分析显示,原发性肝癌患者血清miR-196b表达水平与miR-520f表达水平呈负相关(r=-0.445,P=0.006)。ROC曲线分析显示,血清miR-196b预测原发性肝癌的曲线下面积、截断值、敏感度、特异度及95%置信区间(95%confidence interval,95%CI)分别为0.842,2.55,91.78%,62.50%和95%CI(0.810~0.874);miR-520f预测原发性肝癌的的曲线下面积、截断值、敏感度、特异度及95%置信区间分别为0.872,2.43,91.78%,67.50%和95%CI(0.848~0.906);血清miR-196b联合miR-520f预测原发性肝癌的曲线下面积、敏感度、特异度及95%置信区间分别为0.923,86.30%,87.50%和95%CI(0.891~0.955)。结论 原发性肝癌患者血清miR-196b表达水平上调,miR-520f表达水平下调,其表达水平变化与肿瘤直径、临床分期、分化程度和淋巴结转移密切相关,联合检测血清miR-196b与miR-520f能有效提高原发性肝癌的诊断效能。