Plasma miR‐122 and miR‐200 family are prognostic markers in colorectal cancer

Circulating microRNAs (miRNAs) have been proposed as minimally invasive prognostic markers for various types of cancers, including colorectal cancer (CRC), the third most diagnosed cancer worldwide. We aimed to evaluate the levels of circulating miRNAs that might serve as markers for CRC prognosis and survival. We included plasma samples of 543 CRC patients with stage I‐IV disease from a population‐based study carried out in Germany. After comprehensive evaluation of current literature, 95 miRNAs were selected and measured with Custom TaqMan® Array MicroRNA Cards. Plasma samples of non‐metastatic and metastatic colon cancer patients, each group consisting of ten patients with ‘good’ and ten patients with ‘bad’ prognosis were screened. Identified candidate miRNAs were further validated by RT‐qPCR in the whole study cohort. The association of the miRNA levels with patients' survival and the prognostic subtypes was analyzed with uni‐ and multivariate logistic regression and Cox proportional hazards regression models. Increased miR‐122 levels were associated with a ‘bad’ prognostic subtype in metastatic CRC (Odds ratio: 1.563, 95% confidence interval (CI): 1.038‐2.347) and a shorter relapse‐free survival and overall survival for non‐metastatic (Hazard ratio (HR): 1.370, 95% CI: 1.028‐1.825; HR: 1.353, 95% CI: 1.002‐1.828) and metastatic (HR: 1.264, 95% CI: 1.050‐1.520; HR: 1.292, 95% CI: 1.078‐1.548) CRC patients. Additionally, several members of the miR‐200 family showed associations with patients' prognosis and correlations to clinicopathological characteristics. The here identified miRNA markers, miR‐122 and the miR‐200 family members, could be of use in the development of a multi‐marker blood test for CRC prognosis.     

小鼠巨细胞病毒即刻早期基因M122酵母双杂交诱饵载体的构建及自激活作用的检测

目的构建小鼠巨细胞病毒(MCMV)即刻早期基因M122的酵母双杂交诱饵载体pGBKT7-M122,并检测其自激活作用及对酵母AH109菌株有无毒性作用,用于筛选小鼠胚胎脑cDNA文库。方法采用反转录(RT)-PCR的方法扩增MCMV的M122基因片段,并将其插入到pMD18-Tsimple vector,构建重组质粒pMD18-T-M122。用限制性内切酶EcoRⅠ和SalⅠ将重组质粒酶切鉴定,并测序。测序正确的pMD18-T-M122重组质粒和载体pGBKT7-BD分别用限制性内切酶EcoRⅠ和SalⅠ进行双酶切,凝胶回收M122基因片段及pGBKT7-BD载体片段。将回收的M122基因片段和pGBKT7-BD载体片段,用T4 DNA连接酶16℃连接过夜,构建诱饵质粒pGBKT7-M122。对pGBKTT-M122进行酶切鉴定,测序。将测序正确的pGBKT7-M122转化AH109酵母感受态细胞,转化菌液涂布于营养缺陷培养基SD/-Trp平板和SD/-Trp/X-α-Gal平板。空载体质粒pGBKT7-BD和阳性质粒pCL作为对照亦被转入AH109酵母感受态细胞,转化菌液分别涂布于SD/-Trp平板和SD/...     

Esophagoprotective Potential of Cisapride (An Additional Benefit for Gastroesophageal Reflux Disease)

Cisapride is a novel prokinetic agent thatreleases acetylcholine at the level of the myentericplexus. Acetylcholine also plays a role in the secretoryfunction of salivary glands evoked by intraesophagal mechanical and chemical stimulation, mediatedthrough the esophagosalivary reflex. The impact,however, of cisapride on salivary protective componentsmediated by esophagosalivary reflex remains unknown. Therefore, we have studied salivary pH,bicarbonate, nonbicarbonate, glycoconjugate, protein,EGF, TGF-, and PGE₂ before and afterthe administration of cisapride. The study was conductedin 20 asymptomatic volunteers (9 women and 11 men, mean age 36,range 26-52). Salivary secretions were collected underbasal conditions and during masticatory, mechanical, andchemical stimulation before and after four days of cisapride administration (10 or 20 mg fourtimes a day). Cisapride administration resulted in a 45%increase in salivary volume during the basal condition(P < 0.01), a 32% increase during mastication (P < 0.05), a 53% increase during mechanical(P < 0.05), and a 51% increase during chemical (P< 0.01) stimulation. Cisapride administrationresulted also in a significant increase in salivaryprotein output (P < 0.05), salivary bicarbonate (P <0.05), and nonbicarbonate buffers (P < 0.05), andsalivary EGF (P < 0.05). Salivary glycoconjugatesignificantly increased only during mechanicalstimulation with the catheter and at the end of the esophageal perfusionprocedure (P < 0.05). Although a similar trend wasalso recorded during the analysis of salivaryPGE₂, this difference did not reachstatistical significance. Salivary pH and TGF- before and after cisaprideadministration remained unchanged. The stimulatoryimpact of cisapride on salivary volume and inorganic(bicarbonate and nonbicarbonate buffers) and organic(protein, glycoconjugate, and EGF) protective componentswould benefit patients with GERD and would also bepotential therapy for xerostomia.     

Circ_DOCK1调节miR-122-5p/PPIB轴对结直肠癌细胞增殖、迁移和侵袭的影响

目的 探讨环状核糖核苷酸_胞质分裂作用因子1（Circ_DOCK1）通过调节微小核糖核苷酸-122-5p（miR-122-5p）/肽酰脯氨基顺反异构酶B（PPIB）轴对结直肠癌细胞增殖、迁移和侵袭的影响。方法 实时荧光定量PCR检测结直肠癌患者癌组织与癌旁组织中的circ_DOCK1、miR-122-5p、PPIB表达水平。取人结直肠细胞SW480,脂质体转染法转染circ_DOCK1干扰质粒（sh-circ_DOCK1）、miR-122-5p抑制物（anti-miR-122-5p）和模拟物（miR-122-5p mimics）,以CCK-8法、划痕试验和Transwell实验检测结直肠癌细胞的增殖、迁移和凋亡情况。以生物信息学和荧光素酶报告基因实验分析circ_DOCK1与miR-122-5p的靶向关系。结果 结直肠癌组织的circ_DOCK1、PPIB mRNA表达水平高于癌旁组织,miR-122-5p表达水平低于癌旁组织。与挽救组和NC-circ_DOCK1组比较,sh-circ_DOCK1组的circ_DOCK1和PPIB mRNA表达量下降,miR-122-5p表达量升高,且细胞转染48 h和72 h的A值及细胞迁移率和穿膜细胞数均降低（P＜0.05）;挽救组和NC-circ_DOCK1组的数据比较差异无统计学意义（P＞0.05）。生物信息学软件预测circ_DOCK1含有与miR-122-5p互补的核苷酸序列。转染miR-122-5p mimics后,circ_DOCK1的野生型荧光素酶报告质粒相对活性降低,转染anti-miR-122-5p后,circ_DOCK1的野生型荧光素酶报告质粒相对转录活性增加（P＜0.05）,circ_DOCK1突变型荧光素酶报告质粒相对活性值变化无统计学意义（P＞0.05）。结论 circ_DOCK1在结肠癌组织中的表达上调,且能通过调节miR-122-5p/PPIB轴影响结直肠癌的增殖、迁移和侵袭,可考虑经circ_DOCK1作为结直肠癌的分子靶向治疗研究方向。     

MicroRNA-122-5p与MicroRNA-199a-5p在EMS患者血清的表达及临床意义

目的 探讨microRNA-122-5p（miR-122-5p）与microRNA-199a-5p（miR-199a-5p）在子宫内膜异位症（EMS）患者血清的表达及临床意义。方法 前瞻性设计,随机选取2016年1月—2017年12月华北石油管理局总医院诊治的EMS患者90例（EMS组）及同期不孕症检查的患者70例（对照组）为研究对象。分别采用实时荧光定量聚合酶链反应（qRT-PCR）及酶联免疫吸附试验（ELISA）检测两组人群血清的miR-122-5p和miR-199a-5p的表达及白细胞介素-6（IL-6）的表达,并分析其与临床特征的相关性。构建受试者操作特征（ROC）曲线。结果 EMS组患者的血清及腹腔液中IL-6、miR-122-5p、miR-199a-5p的表达与对照组比较,差异有统计学意义（P?<0.05）,EMS组均增高。miR-122-5p的表达与IL-6水平（r?=0.578,P?<0.05）、miR-199a-5p表达（r?=0.721,P?<0.05）呈正相关。miR-199a-5p的表达与IL-6水平（r?=0.562,P?<0.05）呈正相关。miR-122-5p和miR-199a-5p检测EMS的敏感性分别为94.7%和92.1%,特异性分别为90.5%和88.6%。结论 血清miR-122-5p和miR-199a-5p在EMS患者中的表达增加,miR-122-5p与miR-199a-5p有成为诊断子宫内膜异位症的生物标志物的潜力。     

Heart Transplantation for Homozygous Familial Transthyretin (TTR) V122I Cardiac Amyloidosis

Heart failure is the usual cause of death in patients with amyloid cardiomyopathy. The commonest form of hereditary cardiac amyloidosis is associated with the Val122Ile variant of transthyretin (TTR), which is carried by 3–4% of the African American population. Here, we report the outcome of the first cardiac transplantation in a patient with TTR V122I. A 59-year-old Caribbean man presented with biventricular failure. Other than previous bilateral carpel tunnel syndrome, he had been well and had no evidence of extracardiac amyloidosis. An endomyocardial biopsy demonstrated amyloid of TTR type. Sequencing of TTR gene indicated homozygosity for V122I. He underwent cardiac transplantation and 3 years later, remains well with no evidence of allograft or systemic amyloid deposition.     

Prognostic value of serum microRNA-122 in hepatocellular carcinoma is dependent on coexisting clinical and laboratory factors

BACKGROUND There is ongoing search for new noninvasive biomarkers to improve management of patients with hepatocellular carcinoma(HCC). Studies, mostly from the Asian-Pacific region, demonstrated differential expression of liverspecific microRNA-122(miR-122) in tissue as well as in sera of patients with hepatitis B virus-and hepatitis C virus-induced HCC.AIM To evaluate prognostic value of miR-122 in patients with HCC in a European population and determine potential factors related to alteration of miR-122 in sera.METHODS Patients with confirmed HCC(n = 91) were included in the study over a two-year period. Patients were characterized according to Child-Pugh score, Barcelona clinic liver cancer(BCLC) staging system, etiology of liver disease, laboratory parameters and overall survival. MiR-122 was measured in sera using TaqMan assay normalized to spiked-in cel-miR-39.RESULTS Serum miR-122 quantity was independent of the Child-Pugh score, the BCLC stage or the underlying etiology. Significant positive correlation was found between miR-122 and alanine aminotransferase(P 0.0001), aspartate aminotransferase(P = 0.0001), alpha-fetoprotein(AFP)(P = 0.0034) and hemoglobin concentration(P = 0.076). Negative correlation was observed between miR-122 level and creatinine concentration(P = 0.0028). AFP, ChildPugh score and BCLC staging system were associated with survival differences.In overall cohort low miR-122 in sera was only associated with a trend for a better overall survival without reaching statistical significance. Subgroup analysis revealed that low miR-122 was significantly associated with better prognosis in patients with advanced cirrhosis(Child-Pugh class B/C), advanced tumor stage(BCLC B/C/D) and normal AFP( 7 ng/mL).CONCLUSION Our results strongly support the value of miR-122 as potential biomarker of liver injury and probably prognosis. Nevertheless, the value of miR-122 in prediction of prognosis of HCC patients was limited to certain patients' subgroups. Since circulating miR-122 may be influenced by impaired renal function, AFP and hemoglobin concentration, those factors need to be considered while interpreting miR-122 level.     

Leptin reduces microRNA-122 level in hepatic stellate cells in vitro and in vivo

Obese patients, often accompanied by hyperleptinemia, are more likely to develop liver fibrosis. Leptin, an adipocyte-derived hormone, augments inflammatory in liver and promotes hepatic stellate cell (HSC) activation (a key step for liver fibrogenesis) and liver fibrosis. microRNA-122 (miR-122) is the most abundant liver-specific miRNA and can attenuate liver fibrosis. This study examined the effect of leptin on miR-122 level in HSCs in vivo and in vitro. Results demonstrated that leptin reduced the levels of both miR-122 (mature miR-122) and primary miR-122 (pri-miR-122). The effects of leptin on the levels of miR-122 and pri-miR-122 were through at least hedgehog pathway. Leptin-induced decrease in sterol regulatory element-binding protein-1c (SREBP-1c) has been shown to contribute to leptin-induced HSC activation. We revealed a mutual promotional effect between SREBP-1c and miR-122. Further experiments indicated that miR-122 inhibited leptin-induced liver fibrosis in leptin-deficient mouse model. These data have potential implications for clarifying the mechanisms of hepatic fibrogenesis associated with elevated leptin level in human such as obese patients