Down-regulation of microRNA-181b is a potential prognostic marker of non-small cell lung cancer

The aim of this study was to investigate the clinical significance of microRNA-181b (miR-181b) expression in non-small cell lung cancer (NSCLC).     

miR-181b慢病毒载体的构建及其对HEK-293T细胞MAPK1基因的靶向调控作用

目的构建miR-181b慢病毒过表达载体,探讨其对HEK-293T细胞MAPK1基因的靶向调控作用。方法分别构建p SicoR-miR-181b及pmiR-Report-MAPK1过表达载体,转染HEK-293T细胞后,用实时荧光定量PCR(RTq PCR)法检测MAPK1 mRNA的表达、用Western blot检测MAPK1蛋白的表达及用双荧光素酶报告基因活性验证miR-181b与MAPK1的靶向调控关系。结果成功构建了p SicoR-miR-181b及pmiR-Report-MAPK1重组质粒,miR-181b过表达明显抑制HEK-293T细胞MAPK1蛋白及mRNA表达水平(P0.05),抑制miR-181b的表达后,MAPK1的蛋白及mRNA的表达水平上调(P0.05)。结论成功构建p SicoR-miR-181b慢病毒载体,并证实通过靶向作用MAPK1-3'UTR的特异序列直接抑制MAPK1基因的表达。     

BRAF基因突变在甲状腺乳头状癌中的临床意义

目的：探讨BRAF V600E基因突变与甲状腺乳头状癌的临床相关性。方法：采用PCR技术和直接测序法,对63例甲状腺乳头状癌患者和45例其他类型甲状腺疾病患者石蜡组织BRAF V600E基因突变进行检测。结果：甲状腺乳头状癌组中32例存在突变;而在其他类型甲状腺疾病组中未发现突变。BRAF V600E基因突变在甲状腺乳头状癌不同肿瘤直径组之间的比较差异以及在有无区域淋巴结转移组之间的比较差异均具有统计学意义(P<0.05)。结论：BRAF V600E基因突变与甲状腺乳头状癌的发生、肿瘤的大小和区域淋巴结的转移有关。     

Expression of COX-2 is increased with age in papillary thyroid cancer

AIMS: To study cyclooxygenase-2 (COX-2) and matrix metalloproteinase-2 (MMP-2) expression in papillary thyroid cancer (PTC). Expression of COX-2 is elevated in various human tumours and it has an important role in carcinogenesis. MMP-2 is also an important component of the metastatic potential of tumours. In PTC the most important factor affecting survival is age, but it is poorly understood why older PTC patients have a worse prognosis. METHODS AND RESULTS: This retrospective study comprised 108 patients with PTC, and we compared patients who were either younger than 35 (n = 59) or older than 55 (n = 49). Paraffin-embedded tumour samples were analysed for COX-2 and MMP-2 protein expression using immunohistochemistry. High (scores 2-3) COX-2 immunostaining was observed in 38/108 (35%) of the tumours, and COX-2 expression was significantly (P = 0.002) higher in the older age group (25/49; 51%) than in the young one (13/59; 22%). CONCLUSIONS: Our study shows that COX-2 expression increases with age. It is possible that the age-related increase in COX-2 expression could explain the more aggressive behaviour of PTC in the older age group compared with the young one.     

miR-126 regulated breast cancer cell invasion by targeting ADAM9

Accumulating evidence has shown that microRNAs (miRNAs) deregulation is commonly observed in human malignancies and crucial to cancer metastasis. Herein, we demonstrated that miR-126 play a suppressor role in human breast cancer cells invasion through the direct repression of a disintegrin and metalloprotease 9 (ADAM9). MiR-126 expression was investigated in forty cases of breast cancer specimens by real-time PCR. Transwell assay was conducted to explore the effects of miR-126 on the invasion of human breast cancer cell lines. The impact of miR-126 overexpression on putative target ADAM9 was subsequently confirmed by Western blot analysis. Our results indicated that miR-126 expression was frequently down-regulated in breast cancer specimens compared with adjacent normal tissues (P<0.05). Overexpression of miR-126 significantly reduced (P<0.05) the protein levels of ADAM9, further suppressed (P<0.05) breast cancer cell invasion in vitro. Meanwhile, knockdown of ADAM9 by small interfering RNA (siRNA) also inhibited (P<0.05) breast cancer cell invasion. Thus, our study revealed that miR-126 may act as a tumor suppressor via inhibition of cell invasion by downregulating ADAM9 in breast cancer development.     

microRNA-181b suppresses the metastasis of lung cancer cells by targeting sex determining region Y-related high mobility group-box 6 (Sox6)

Background

The aim of the study was to measure the expression of microRNA (miR)-181b in patients with lung cancer, investigate its biological function and elucidate the underlying mechanisms associated with the development of lung cancer.

Pitfalls in the surgical pathologic diagnosis of papillary thyroid carcinoma

While the surgical pathology examination of the thyroid gland for papillary carcinoma may seem, on its surface, to be relatively straightforward, in reality it is fraught with diagnostic traps. Avoidance of these pitfalls is necessary for guiding the surgeon and endocrinologist to the appropriate treatment and follow up. This review will detail a selected group of some of the more commonly encountered challenges in making the diagnosis of papillary thyroid carcinoma from a busy head and neck pathology consultation practice.     

Macrofollicular variant of papillary thyroid carcinoma

A rare case of a macrofollicular variant of papillary thyroid carcinoma occurring in an 18-years-old male is described. The extirpated tumor, 5.5 × 5.5 × 3.5 cm In size, was well demarcated and multinodular, and histopathologically showed a predominantly macrofolllcular structure reminiscent of adenomatous goiter or macrofollicular adenoma. In the tumor tissue, however, there were several small foci of microfollicular or paplllary structure with the nuclei charae teristic of papillary carcinoma. Parts of the macrofollicular areas also showed similar nuclear characteristics with a transition to microfollicular or papillary areas. Incomplete capsular invasion and minimal vascular invasion were also present. Additional resected specimens contained small metastatic nodules in the residual left lobe and lymph nodes. Immunohistochemistry showed a small number of p53-positive tumor cells in the microfollicular or papillary areas. It is suggested that this tumor is a well-differentiated variant which should be distinguished from benign thyroid lesions, although there have been some cases of metastases which appear related to capsular and/or vascular invasion.     

Down-regulation of microRNA-135b inhibited growth of cervical cancer cells by targeting FOXO1

More and more evidence has confirmed that dysregulation of microRNAs (miRNAs) can conduce to the progression of human cancers. Previous studied have shown that dysregulation of miR-135b is in varieties of tumors. However, the roles of miR-135b in cervical cancer remain unknown. Therefore, our aim of this study was to explore the biological function and molecular mechanism of miR-135b in cervical cancer cell lines, discussing whether it could be a therapeutic biomarker of cervical cancer in the future. The MTT assay and ELISA-Brdu assay were used to assess cell proliferation. Cell cycle was detected by flow cytometry. Real-time quantitative polymerase chain reaction (PCR) and Western blot analyses were used to detect expressions of cyclin D1, p21, p27 and FOXO1. In our study, we found that miR-135b is up-regulated in cervical cancer cell lines. Down-regulation of miR-135b evidently inhibited proliferation and arrested cell cycle in cervical cancer cells. Bioinformatics analysis predicted that the FOXO1 was a potential target gene of miR-135b. Besides, miR-135b inhibition significantly increased expressions of the cyclin-dependent kinase inhibitors, p21^/CIP1 and p27^/KIP1, and decreased expression of cyclin D1. However, the high level of miR-135b was associated with increased expression of FOXO1 in cervical cancer cells. Further study by luciferase reporter assay demonstrated that miR-135b could directly target FOXO1. Down-regulation of FOXO1 in cervical cancer cells transfected with miR-135b inhibitor partially reversed its inhibitory effects. In conclusion, down-regulation of miR-135b inhibited cell growth in cervical cancer cells by up-regulation of FOXO1.     

miR-193b体外协同增强多柔比星对乳腺癌细胞的抗肿瘤效应

目的:研究微小RNA(microRNA,miR)-193b是否能增强多柔比星对乳腺癌细胞的杀伤效力及机制。方法:用real-time PCR方法检测乳腺癌患者及健康对照者血浆中的miR-193b表达水平。MTT法检测miR-193b联合多柔比星对乳腺癌细胞系MDA-MB-231的杀伤效力。利用生物信息学、real-time PCR及Western blot方法验证miR-193b是否调节乳腺癌细胞Mcl-1的表达。构建Mcl-1真核表达载体,MTT法检测Mcl-1表达载体转染对miR-193b联合多柔比星治疗乳腺癌疗效的影响。结果:乳腺癌患者血浆中miR-193b表达水平显著低于对照组。miR-193b联合多柔比星治疗组对MDA-MB-231细胞的杀伤效力显著高于多柔比星单治疗组。miR-193b转染后,MDA-MB-231细胞Mcl-1的mRNA及蛋白表达水平均下降。miR-193b联合多柔比星在Mcl-1表达载体转染后对MDA-MB-231细胞的杀伤活性显著低于未转染Mcl-1表达载体的miR-193b联合多柔比星组。结论:miR-193b通过靶向于Mcl-1增强多柔比星对乳腺癌细胞的杀伤效力。     

Galectin-3 expression in papillary microcarcinoma of the thyroid

AIMS: Galectin-3 is a beta-galactoside binding protein, recently recognized as a promising molecular marker of thyroid malignancy. As reported in several studies, galectin-3 is highly expressed in papillary thyroid carcinoma, but its expression has not been investigated in papillary microcarcinoma, which is a variant of papillary thyroid carcinoma. METHODS AND RESULTS: Using a monoclonal antibody to galectin-3 and the avidin-biotin-peroxidase complex (ABC) immunohistochemical technique, we analysed galectin-3 expression in 63 cases of papillary microcarcinoma. The results showed immunohistochemical reactivity for galectin-3 in 51 (80.9%) cases. Intensity of staining varied from strong or moderate to weak. Galectin-3 localization was mostly cytoplasmic, but also membranous or nuclear in some cells. Immunohistochemical expression of galectin-3 was not found in 12 (19.1%) cases. Most galectin-3 negative microcarcinomas (10/12) were of the non-classical type, i.e. without papillary architecture. Neither the frequency nor the intensity of a positive reaction was related to tumour size. CONCLUSIONS: Galectin-3 gene is expressed at the protein level in most papillary microcarcinomas, although with slightly lower frequency than that reported for clinically evident papillary thyroid carcinoma. The presence of galectin-3 in clinically silent microcarcinomas may indicate that galectin-3 is not related to growth or aggressiveness of papillary thyroid microcarcinomas but rather plays some other role in thyroid tumour biology.     

甲状腺乳头状癌核内包涵体及核的形态结构定量分析

目的:定量揭示甲状腺乳头状癌核内包涵体及核的几何形态结构特点,为甲状腺乳头状癌的病理诊断提供定量诊断参数。方法:取甲状腺乳头状癌100例,常规切片、HE染色,在40倍物镜下采集核内包涵体及核的图像,用Image-Pro Plus测试包涵体及核的面积、周长、长轴、短轴、形状因子PE（Form PE）、形状因子AR（Form AR）、规化形状因子（RFF）、轴比、核内包涵体的面积密度以及包涵体与核质的面积比。结果:（1）甲状腺乳头状癌核内包涵体的面积约为（13.575±9.045） μm2（95%CI 13.339~13.810）,其在核内的面积密度约为25.761%±10.683%（95%CI 25.483~26.039）;其周长约为（12.720±4.275） μm（95%CI 12.609~12.831）,长轴（4.547±1.466） μm（95%CI 4.509~4.585）,短轴（3.482±1.178） μm （95%CI 3.451~3.512）;其轴比约为0.772±0.121（95%CI 0.769~0.775）,Form PE、Form AR和RFF约等于1;其与核质的面积比约为0.380±0.237（95%CI 0.374~0.386）,其面积、周长、长轴和短轴的变异系数为66.6%、33.6%、32.2%、33.8%。（2）具有核内包涵体的癌细胞,其核的面积、周长、长轴和短轴及其相应变异系数显著大于没有核内包涵体的细胞核（P<0.05）。（3）核内包涵体、含有核内包涵体的核及没有核内包涵体的核,其面积、周长、长轴和短轴的频数分布均呈正偏态分布。结论:（1）甲状腺乳头状癌核内包涵体形状大致呈圆形;约为核面积的1/4,核周长、长轴和短轴的1/2;其面积、周长、长轴和短轴的变异程度均较大。（2）具有包涵体的癌细胞,其核的异型性明显,诊断阅片时应重点观察。（3）包涵体及核的上述尺寸参数的频数分布均呈正偏态分布。     

MiR-125b promotes migration and invasion by targeting the vitamin D receptor in renal cell carcinoma

Purpose: To investigate the expression of miR-125b and vitamin D receptor (VDR) in renal cell carcinoma (RCC) and assess the biological function of miR-125b in RCC.Methods: We used quantitative real-time polymerase chain reaction (RT-PCR) to detect the expression of nucleic acids and western blotting to analyze the protein abundance in RCC cell lines. MiR-125b mimic and inhibitor were employed to investigate the function and behavior of miR-125b in RCC cell lines. The relationship between miR-125 and VDR was verified using luciferase assays.Results: Overexpression of miR-125b promoted migration and invasion and prevent cell apoptosis in ACHN cells. In contrast, miR-125b deficiency suppressed migration and invasion and induced cell apoptosis in 786-O cells. Luciferase assays indicated the interaction between miR-125b and VDR. In collected samples, miR-125b was significantly higher in RCC tissues and negatively correlated to VDR (r=-0.444, p=0.04).Conclusion: MiR-125b displays an oncogene profile in RCC, patients with high expression of miR-125b should be a more frequent follow-up. MiR-125B may be a potential therapeutic target for RCC.     

下调microRNA-181b在小鼠缺血性脑损伤中的神经保护作用

目的:探讨microRNA-181b(miR-181b)在缺血性小鼠脑损伤病理过程中的神经保护作用。方法:应用N2A细胞氧-糖剥夺(OGD)模型模拟神经细胞缺血损伤;应用小鼠大脑中动脉阻塞(MCAO)模型模拟缺血性脑损伤(成功率约60%),原位细胞凋亡检测试剂盒检测N2A细胞损伤程度,蛋白印迹检测miR-181b靶蛋白自噬蛋白5(Atg5)及caspase-9的变化情况,萤光素酶报告基因技术检测miR-181b对Atg5 mRNA的直接调控作用。结果:在OGD致N2A细胞缺血损伤过程中,通过上调或抑制miR-181b的表达水平可以显著影响N2A细胞的凋亡(P0.05);miR-181b表达水平的改变可显著影响Atg5蛋白表达水平(P0.05);共转染miR-181b前体或miR-181b抑制剂可显著抑制或增高含有Atg5 mRNA 3’-UTR的萤光素酶报告基因的活性(P0.05);MCAO后活化caspase-9的表达显著升高,而miR-181b拮抗剂可使MCAO后剪切的caspase-9表达明显减少(P0.05)。结论:下调miR-181b可能通过调节Atg5的蛋白表达在缺血性小鼠脑损伤中发挥神经保护作用。