FOXM1及E-钙黏蛋白在乳腺癌组织中的表达及其临床意义的研究

目的探讨乳腺癌病人根治性术后癌组织、非癌细胞浸润组织中FOXM1和E-钙黏蛋白的表达及与病理参数的关系和临床意义。方法免疫组织化学方法(IHC)检测术后乳腺癌及非癌细胞浸润组织中FOXM1和E-钙黏蛋白的表达,利用SPSS软件分析其与临床病理特征的关系。结果 68例乳腺浸润性导管癌组织中,FOXM1蛋白的阳性表达率为48. 5%(33/68),E-钙黏蛋白的阳性表达率为41. 2%(28/68)。肿瘤组织中FOXM1蛋白的表达高于非癌细胞浸润组织。FOXM1和E-钙黏蛋白的阳性表达与病人年龄、部位、组织学分型、分子分型均无关。FOXM1在腋窝淋巴结转移组中其阳性表达率高于无淋巴结转移组(P 0. 05),并与肿瘤大小、肿瘤TNM分期、雌激素受体状态相关(P 0. 05),E-钙黏蛋白的表达率降低与淋巴结转、肿瘤TNM分期密切相关(P 0. 05)。FOXM1和E-钙黏蛋白的表达呈负相关(r=-0. 324,P 0. 05)。结论 FOXM1和E-钙黏蛋白是乳腺癌浸润、转移的重要的生物学标记物。     

FOX家族基因在肿瘤形成中的作用研究进展

转录因子FOX家族基因具有重要的生物学功能,包括调控细胞增生和分化以及肿瘤形成,其表达失调后具有致癌基因和抑癌基因的功能,相关研究已越来越受到关注.本文就目前关于FOX家族基因在相关肿瘤形成中的作用研究作一综述.     

同源框基因在消化道肿瘤中的异常表达及相关机制研究

同源框基因不仅主控细胞增生和分化,而且还参与调节人类多种恶性肿瘤的发生、发展及预后.近来有研究显示同源框基因在食管癌、胃癌、大肠癌等消化道肿瘤中异常表达,深入研究后发现同源框基因可能通过体内相关转录因子及表现遗传异常修饰等途径,调探消化道肿瘤的发生、发展.因此,同源框基与消化道肿瘤密切相关.     

MTA1与肿瘤转移的研究进展

MTA1是Nurd的组成成分,它通过促进组蛋白、非组蛋白去乙酰化影响基因的转录和蛋白的表达.MTA1在肿瘤组织中异常表达,与肿瘤的侵袭、转移和预后密切相关,具有重要的临床应用价值.本文对MTA1的结构、功能及其在多种肿瘤转移中的研究进展进行了综述.     

DNA甲基化转移酶1/3b与肝癌临床病理特征和预后相关性研究

目的探讨DNA甲基化转移酶1/3b(DNMT1/3b)表达异常与肝癌临床病理特征和患者术后生存之间的相关性。 方法采用免疫组织化学技术检测89例手术切除肝癌组织中DNMT1和DNMT3b的蛋白表达情况,结合临床病理及无瘤生存期和总生存期,分析DNMT1/3b与临床病理参数、预后的相关性,并通过高通量基因芯片技术筛选肝癌相关DNMT1和DNMT3b的靶基因。 结果89例肝癌组织中DNMT1和DNMT3b皆为阳性63例,皆为阴性5例,单一阳性的分别为9例和12例。DNMT1/3b在肝癌组织中高表达与患者血清AFP(χ²=12.903,P=0.005)、乙型肝炎(χ²=9.535,P=0.023)、卫星灶(χ²=9.574,P=0.023)和肿瘤复发、转移及生存期有关,而与性别、肿瘤大小、肝硬化和肿瘤分化无关;Kaplan-Meier分析显示DNMT1/3b阳性组患者生存时间较阴性组短,差异有统计学意义(P<0.05)。高通量基因组甲基化芯片技术筛选出肝癌细胞中受DNMT1和DNMT3b调控的基因2 000多个,其中参与肿瘤侵袭转移的靶基因有112个。 结论DNMT1/3b在肝癌组织中高表达与患者血清AFP、乙型肝炎、卫星灶和肿瘤复发、转移等临床病理特征和生存期密切相关,并可能通过调控多方面肿瘤相关基因促进肝癌复发、转移等恶性表型。     

肿瘤转移相关基因MTA1与鼻咽癌细胞浸润和转移的关系

目的:鼻咽癌是高转移和高侵袭性肿瘤,而且对现行的治疗方法有抵抗性.肿瘤转移相关基因MTA1是新近发现的肿瘤转移相关基因.本实验MTA1基因在人鼻咽癌细胞高低转移株5-8F和6-10B的表达水平以及低转移细胞株转染MTA1全长基因后侵袭能力的改变,探讨MTA1表达与鼻咽癌侵袭转移的相关性.方法:两株细胞株中的MTA1表达通过半定量的RT-PCR进行检测,用细胞侵袭实验及细胞增值实验评价两株细胞株离体的增值及侵袭能力,并检测低转移株(6-10B)转染MTA1CDNA后MTA1的表达及侵袭能力的变化.结果:高转移株5-8F中的MTA1的表达明显高于低转移株6-10B呈正相关,而且低转移株6-10B转染MTA1后其MTA1的表达及侵袭能力明显增加.结论:在鼻咽癌细胞中MTA1同肿瘤的侵袭转移能力呈明显相关性,MTA1可以作为研究鼻咽癌转移及治疗的候选基因.     

Pokemon基因的研究进展

原癌基因Pokemon又叫ZBTB7、LRF、FBI-1,其编码产物属于POK家族,是一种转录抑制因子.研究认为它调控多种肿瘤相关基因的转录,是肿瘤发生发展过程中的上游关键基因.目前已在人类多种肿瘤的研究中发现了Pokemon的异常表达.近年的一些文献强调了这个转录抑制因子在癌症发生、细胞生理和干细胞生物学中的重要性....     

Maspin:新的潜在的骨肉瘤转移抑制基因

目的 通过对肿瘤基因组数据整体分析寻找骨肉瘤转移相关基因,初步分析其与骨肉瘤转移的关系.方法 联合分析肿瘤组织DNA拷贝数目的 变化及肿瘤转移细胞模型中表达差异基因数据,筛选骨肉瘤转移相关基因,通过RT-PCR、Westen blot及免疫组化检测该基因在不同骨肉瘤细胞系、正常成骨细胞、骨肉瘤及正常骨组织临床标本中的表达,并结合临床随访分析其与骨肉瘤患者预后的关系.结果 NCBI数据库显示18q常出现DNA拷贝数目变化和杂合性缺失.基因芯片结果 显示高转移MG63-A1细胞系与MG63wt相比142个基因表达差异在4倍以上,而在18q21区域其中一个新的肿瘤转移抑制基因Maspin在高转移MG63-A1细胞系中表达明显下降.RT-PCR、Westen blot检测证实基因芯片结果 准确,并发现Maspin在正常成骨细胞高表达,在SAOS2、143B中表达缺失.免疫组化检测显示在27例骨肉瘤临床标本中,33.3%(9/27例)的标本Maspin表达为阳性,其余为阴性;在正常骨组织中Maspin表达强阳性.Maspin表达与骨肉瘤患者预后呈正相关(P=0.032).结论 Maspin的表达随着骨肉瘤细胞系转移能力的增加而出现缺失,在骨肉瘤标本中Maspin表达与患者预后呈正相关,预示着其可能成为阻遏骨肉瘤转移的新靶点.     

MALAT1在结直肠恶性肿瘤诊治中的作用

<正>结直肠癌发病率居高不下,我国每年有超过90万人死于结直肠癌^[1]。结直肠癌与饮食、遗传、炎症等因素密切相关,部分患者在确诊前已发生转移,预后极差^[2]。目前结直肠癌具体发病机制尚不明确。近些年来,随着长链非编码RNA(LncRNA)的分子生物学功能被不断挖掘,大量研究证实其与肿瘤的发生发展密切相关,可通过多种方式参与肿瘤细胞的转移、增殖等^[3-4]。肺腺癌转移相关转录本1(MALAT1)作为LncRNA中的一员,起初在非小细胞肺癌的研究中被发现,随后被证实在多种肿瘤进展中均存在差异性表达,从表观遗传、转录及转录后水平等多方面参与基因调控,影响肿瘤病理生理过程^[5]。研究发现MALAT1过表达对肿瘤的侵袭及转移发挥促进作用^[6]。本文就MALAT1在结直肠恶性肿瘤诊治中的作用进行简要介绍。     

乳腺癌转移抑制基因1抑制乳腺癌转移的作用机理研究进展

目的综述乳腺癌转移抑制基因1（BRMS1）在抑制乳腺癌转移中的作用机理研究进展。方法采用文献回顾的方法，对目前国内、外有关BRMS1在乳腺癌中的研究状况加以分析与综述。结果BRMS1与其他肿瘤转移抑制基因一样，主要抑制肿瘤的转移，并不影响肿瘤的生长，在乳腺癌中主要通过调节细胞间的信号转导及其他转移抑制基因的表达而抑制乳腺癌的转移。结论对BRMS1基因的深入研究有助于进一步深化对乳腺癌转移的认识，为肿瘤转移的分子诊断和基因治疗提供新的思路。     

短发卡RNA稳定沉默FOXM1对肝癌细胞体外生长的影响

目的 探讨短发夹RNA(short hairpin RNA,shRNA)表达载体稳定沉默叉头框M1(forkhead box M1,FOXM1)基因对人肝癌细胞体外生长的影响.方法 构建针对人类FOXM1mRNA的不同干扰靶点的4个shRNA表达载体,选择干扰效果最佳的表达载体和阴性对照质粒转染人肝癌细胞QGY-7703,用新霉素(G418)筛选稳定转染的克隆.用四甲基偶氮唑盐(MTT)比色法和平板克隆形成实验检测FOXM1基因沉默前后肝癌细胞体外生长能力的变化.用Annexin V-APC/PI双染法检测细胞凋亡.结果 不同人肝癌细胞株中普遍表达FOXM1蛋白.4个shRNA表达载体中,shRNA-1026表达载体的干扰效果最佳,对FOXM1 mRNA和蛋白表达水平的抑制率分别为38.5%和53.2%.用shRNA-1026稳定沉默FOXM1基因后,QGY-7703细胞增殖受到抑制,培养48、72和96 h后,沉默组的吸光值均显著低于对照组(分别t=10.830,3.578,5.734,均P＜0.05);沉默组的克隆形成能力较对照组显著降低(t=5.336,P＜0.05),而细胞凋亡较对照组显著增加(t=6.827.P＜0.05).结论 shRNA表达载体稳定沉默FOXM1基因抑制肝癌细胞的生长.

Abstract：

Objective To evaluate the effect of sustained silencing Forkhead box Ml (F0XM1) gene by short-hairpin RNA (shRNA) expression vector on cell growth of hepatocelluar carcinoma (HCC) in vitro.Methods Four shRNA expression vectors targeting different sequences of human F0XM1 mRNA were constructed.The expression vector with the best interfering effect and the negative control plasmid were used to transfect HCC cell line QGY-7703, stably transfected cell clones were selected by neomycin (G418).Cell growth was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and colony formation was assessed by clonogenic assay.Cell apoptosis was detected by double staining with APC conjugated Annexin V and PI.Results F0XM1 protein was detected with different levels in all these studied human cell lines.The expression vector shRNA-1026 exhibited excellent interference effect after transient transfection, which showed 38.5% and 53.2% reduction of FOXM1 mRNA and protein level respectively.The growth of QGY-7703 cells was inhibited after stable inhibition of FOXM1 expression by shRNA-1026, which was indicated by decreased absorbance value of the test group after culture for 48, 72 and 96 h compared to control group (t = 10.830,3.578 and 5.734 respectively, P ＜ 0.05).Stable inhibition of F0XM1 also led to reduced colony formation ( t = 5.336, P ＜ 0.05 ) and increased apoptosis of QGY-7703 cells in comparison to control cells (t = 6.827, P ＜ 0.05 ).Conclusions Stable silencing F0XM1 gene by shRNA suppresses the growth of HCC cells in vitro.     

原发性肝癌术中化疗泵埋置途径选择的研究

目的　探讨原发性肝癌 (PHC)病灶在不同切除方式时化疗泵埋 (DDS)置途径的选择。方法　将 190例原发性肝癌患者按手术切除方式分为原发肿瘤完整切除 (A)、未完全切除 (B)组两组。再根据肿瘤不同切除方式时所采用置泵途径 ,两大组各分为经肝动脉 门静脉双途径置泵 (a)组、经门静脉 (b)组、经肝固有动脉 (c)三组。结果　 (1)A组中 ,c组的 1年、3年复发率均高于b组 ;a组的 1年、3年复发率与b组无差异。b组与a、c两组相比 ,其生存率与a组无明显差异 ,较c组生存率高。 (2 )B组中 ,a组的 1年、3年生存率与b组有显著差别 ;c组预后差于a组 ,但与b组无差别。结论　在原发肿瘤完整切除的情况下 ,首选经门静脉置泵 ;在原发肿瘤未能完整切除的患者 ,只能选择双途径置泵。     

The upregulation of osteoblast marker genes in mesenchymal stem cells prove the osteoinductivity of hydroxyapatite/tricalcium phosphate biomaterial

Background

Calcium phosphate (Ca-P), mainly concerning hydroxyapatite (HA), is the main inorganic component of the body's hard tissue. It is acknowledged that Ca-P biomaterial not only has osteoconduction but also can form bone bonding to host bone, making an ideal tissue-engineering scaffold. However, whether Ca-P biomaterial possesses osteoinductivity is still debated. The present study was performed to explore the expression level of osteoblast maker genes in human mesenchymal stem cells (hMSCs) grown on a Ca-P biomaterial.

Materials and methods

hMSCs were cultured on the HA/tricalcium phosphate-(TCP) double-phase ceramic. After coculture for 5, 10, 15, or 20 days, the cells were digested for isolation of total RNA. Fluorescence quantitative polymerase chain reaction was used to detect the relative mRNA levels of Runx2, collagen type I, osteopontin, and osteocalcin, all of which are the marker genes for osteoblasts. The mg63 cell was recruited as the reference and un-cocultured hMSCs as the negative controls. Alkaline phosphatase (ALP) activity in the cells was also examined after culture for 10 or 20 days.

Results

Our results showed that the expression levels of all four genes continued to rise during the first 10 days. Then, both collagen type I and Runx2 decreased. In contrast, osteocalcin mRNA reached its maximum at day 15 and osteopontin mRNA kept increasing throughout the whole experimental period. Additionally, ALP activity increased in a time-dependent manner.

Conclusion

The up-regulation of all four osteoblast marker genes in hMSCs grown on Ca-P biomaterial suggested that HA/TCP biomaterials possess osteoinductivity on hMSCs, cells a mechanism that requires further investigation.     

Thoracoscopic resection of neurogenic tumors in children

Purpose

The aim of this study was to evaluate the feasibility of thoracoscopy in neurogenic tumors in infants and children.

Materials and Methods

From January 2000 to October 2005, 21 patients aged 7 months to 14 years (mean, 6 years) underwent thoracoscopy for tumor resection in 5 French institutions. One 10-mm optical port and 2 operative 5-mm ports were needed. Selective intubation was required for 3 patients aged about 12 years. Tumor was removed with an endoscopic bag in all cases.

Results

All procedures were completed successfully without any incomplete resection or recurrence. One conversion was necessary because of a huge mass. A chest tube was left for a mean of 2 days for 17 children. Two children had not had any drainage. Two postoperative chylothorax required chest drainage for 12 days. Only 5 of the 6 older patients (mean age, 12 years) needed a patient-controlled analgesia. The mean operative time was about 100 minutes. Hospital stay ranged from 4 to 12 days. Tumors were neuroblastoma or ganglioneuroblastoma in 16 cases and ganglioneuroma in the 5 other cases.

Conclusion

Thoracoscopy for resection of thoracic neurogenic tumors in children is a feasible, safe, and efficient procedure. The surgeon has a better visualization of the tumor and its anatomic connections. Resection can be as complete as an open procedure without having to complicate the operative technique in the same operating time. It avoids cosmetic and functional disorders because of thoracotomy. It allows a good cosmetic resection without spillage.     

Independent validation of the 2002 American Joint Committee on cancer primary tumor classification for renal cell carcinoma using a large, single institution cohort

PURPOSE: The primary tumor classification for renal cell carcinoma (RCC) was updated by the American Joint Committee on Cancer in 2002. To date the new classification has not been validated using an independent group of patients and, therefore, its accuracy for predicting patient outcome is unknown. In the current study we evaluated the 2002 primary tumor classification and compared its predictive ability with that of the 1997 classification. MATERIALS AND METHODS: We studied 2,746 patients treated with radical nephrectomy or nephron sparing surgery for unilateral, sporadic RCC between 1970 and 2000. Cancer specific survival was estimated using the Kaplan-Meier method. The predictive abilities of the 1997 and 2002 classifications were compared using the concordance index. RESULTS: There were 812 deaths from RCC a mean of 3.3 years following nephrectomy. Median followup in patients still alive at last followup was 9 years. Estimated 5-year cancer specific survival rates by the 2002 tumor classification were 97%, 87%, 71%, 53%, 44%, 37% and 20% in patients with pT1a, pT1b, pT2, pT3a, pT3b, pT3c and pT4 RCC, respectively. The concordance index for the association between the 2002 classification and death from RCC was 0.752 compared with 0.737 for the 1997 classification, indicating that the 2002 version contained more predictive ability. CONCLUSIONS: Our data suggest that the 2002 primary tumor classification with pT1 cancers subclassified into pT1a and pT1b provides excellent stratification of patients according to cancer specific survival and it has a predictive ability that is superior to that of the 1997 classification.     

Expression of micro-RNAs and genes related to angiogenesis in ccRCC and associations with tumor characteristics

Background

Clear cell renal cell carcinoma (ccRCC) is the third most common urological cancer in adults. Our aim is to evaluate genes and miRNAs expression profiles involved with angiogenesis and tumor characteristics in ccRCC.

Methods

The expression levels of miRNAs miR-99a, 99b, 100; 199a; 106a; 106b; 29a; 29b; 29c; 126; 200a, 200b and their respective target genes: mTOR, HIF1-α, VHL, PDGF, VEGF, VEGFR1 and VEGFR2 were analyzed using qRT-PCR in tumor tissue samples from 56 patients with ccRCC. Five samples of benign renal tissue were utilized as control. The expression levels of miRNAs and genes were related to tumor size, Fuhrman nuclear grade and microvascular invasion.

Results

miR99a was overexpressed in most samples and its target gene mTOR was underexpressed, this also occurs for miRNAs 106a, 106b, and their target gene VHL. An increase in miR-200b was correlated with high-risk tumors (p?=?0.01) while miR-126 overexpression was associated with Fuhrman’s low grade (p?=?0.03).

Conclusions

Our results show that in ccRCC there are changes in miRNAs expression affecting gene expression that could be important in determining the aggressiveness of this lethal neoplasia.

     

miR-30 Promotes Thermogenesis and the Development of Beige Fat by Targeting RIP140

Members of the microRNA (miR)-30 family have been reported to promote adipogenesis and inhibit osteogenesis, yet their role in the regulation of thermogenesis remains unknown. In this study, we show that miR-30b/c concentrations are greatly increased during adipocyte differentiation and are stimulated by cold exposure or the β-adrenergic receptor activator. Overexpression and knockdown of miR-30b and -30c induced and suppressed, respectively, the expression of thermogenic genes such as UCP1 and Cidea in brown adipocytes. Forced expression of miR-30b/c also significantly increased thermogenic gene expression and mitochondrial respiration in primary adipocytes derived from subcutaneous white adipose tissue, demonstrating a promoting effect of miRNAs on the development of beige fat. In addition, knockdown of miR-30b/c repressed UCP1 expression in brown adipose tissue in vivo. miR-30b/c targets the 3′-untranslated region of the receptor-interacting protein 140 (RIP140), and overexpression of miR-30b/c significantly reduced RIP140 expression. Consistent with RIP140 as a target of miR-30b/c in regulating thermogenic gene expression, overexpression of RIP140 greatly suppressed the promoting effect of miR-30b/c on the expression of UCP1 and Cidea in brown adipocytes. Taken together, the data from our study identify miR-30b/c as a key regulator of thermogenesis and uncover a new mechanism underlying the regulation of brown adipose tissue function and the development of beige fat.