特异AT序列结合蛋白1在卵巢癌的表达及临床病理学意义

目的:检测上皮性卵巢癌中特异AT序列结合蛋白1(Special AT-rich sequence-bindingprotein 1,SATB1)的表达,探讨SATB1表达与卵巢癌的发生,进展及转移的关系及意义.方法:以实时荧光定量RT-PCR和免疫组化方法检测91例上皮性卵巢癌组织、13例卵巢交界性囊腺瘤及8例正常卵巢组织中SATB1mRNA和蛋白的表达,分析SATB1基因表达与临床病理参数的相关性.结果:上皮性卵巢癌组织和交界性囊腺瘤组织中SATB1 mRNA表达量分别为正常卵巢组织的6.97倍和5.70倍,差异有统计学意义(P＜0.05).SATB1 mRNA表达水平随着卵巢癌分期升高而增加:Ⅰ期,Ⅱ期和Ⅲ期分别为正常卵巢组织的5.31倍,6.67倍和8.04倍(P＜0.05),并且在淋巴结转移组表达明显高于无转移组(P＜0.05).免疫组化也显示SATB1蛋白在上皮性卵巢癌中表达高于正常卵巢组织,差异具有统计学意义(P＜0.05).结论:SATB1的表达水平可能与上皮性卵巢癌发生,进展及转移有关.     

HIF-1α 在卵巢癌中的表达及与化疗耐药相关性研究

目的:探讨缺氧诱导因子-1α(HIF-1α)在卵巢癌中的表达及其在卵巢癌化疗耐药中的作用。方法:采用免疫组化方法,检测96例卵巢癌组织,45例良性卵巢肿瘤和30例正常卵巢组织中HIF-1α的表达,分析其表达与临床病理特征及化疗耐药的相关性。结果:卵巢癌中HIF-1α的表达高于正常卵巢及良性卵巢肿瘤组织,且临床分期越晚其表达越高(P<0.001),而与肿瘤组织类型、病理分级及患者年龄无显著相关性。HIF-1α在卵巢癌化疗耐药组阳性率明显高于化疗敏感组(P<0.001)。结论:卵巢癌中HIF-1α高表达与卵巢癌的发生、发展、浸润和转移有关,与卵巢癌化疗耐药密切相关,HIF-1α可成为卵巢癌化疗新的分子治疗靶点。     

微小RNA-21与PTEN在上皮性卵巢癌组织中的表达及临床意义

目的:检测微小RNA-21(miR-21)和抑癌基因PTEN在上皮性卵巢癌组织中的表达,探讨二者的相关性及其与临床病理特征间的关系.方法:采用茎环实时荧光逆转录聚合酶链反应(Real-time RT-PCR)检测48例上皮性卵巢癌组织、24例良性上皮性卵巢肿瘤及15例正常卵巢组织标本中miR-21表达.采用链霉菌抗生物素蛋白一过氧化酶(SP)免疫组化方法检测上述组织中PTEN蛋白表达,分析mir-21表达和PTEN表达的相关性及其与临床病理特征的关系.结果:茎环Real-time RT-PCR检测miR-21表达的敏感性和特异性良好;miR-21在卵巢癌组织中的相对表达量(4.849±1.813)显著高于良性卵巢肿瘤(1.133±0.291)及正常卵巢组织(P<0.01),PTEN在卵巢癌组织中阳性表达率为(41.66%),明显低于卵巢良性肿瘤(83.33%)及正常卵巢(100%).卵巢癌组织中miR-21表达量与PTEN阳性表达率存在显著负相关(r=-0.447,P<0.01).miR-21及PTEN表达随卵巢癌的临床分期和组织分级的进展分别呈上升及下降趋势,而且二者表达均与淋巴结转移相关(P均<0.01),而与组织病理类型无关.结论:miR-21在上皮性卵巢癌组织中的表达升高,可能通过负性调节PTEN表达在卵巢癌的发生发展中发挥致癌基因的作用,并且与卵巢癌的演进及不良预后密切相关.     

Skp2和P27在上皮性卵巢癌中表达及其临床意义

目的:检测Skp2和P27在上皮性卵巢癌中的表达,分析Skq2和P27与临床病理特征间的关系,探讨二者在上皮性卵巢癌发生发展中的作用.方法:应用免疫组织化学SP法,检测69例上皮性卵巢癌、15例良性卵巢肿瘤以及15例正常卵巢组织中Skp2和P27的表达,分析Skp2和P27表达与上皮性卵巢癌临床病理参数的关系,及两者的相关性.结果:上皮性卵巢癌、良性卵巢肿瘤及正常卵巢组织Skp2和P27表达阳性率各为42.02%,26.67%、6.67%和47.82%,66.67%、86.67%,差异有统计学意义(P<0.05).Skp2的表达与临床分期和病理组织学分级显著相关,与患者年龄、病理类型、有无淋巴结转移和肿瘤大小无显著相关.P27的表达与临床分期、有无淋巴结转移和病理组织学分级显著相关,与患者年龄、病理类型和肿瘤大小无显著相关.Skp2和P27在上皮性卵巢癌中表达呈显著负相关,相关系数r=-0.463,(P<0.01).结论:Skp2与P27呈负相关,二者表达可能与上皮性卵巢癌的发生发展有关.     

p27kip1、Cyclin D1在卵巢癌中的表达及临床意义

目的探讨p27kip1、Cyclin D1在卵巢癌发生方面的意义.方法应用免疫组织化学方法及半定量分析方法,检测50例卵巢癌、19例卵巢良性上皮肿瘤、 13例正常卵巢组织中的p27kip1和Cyclin D1表达,并分析它们与良恶性肿瘤、病理学分级、临床分期的相关性. 结果正常卵巢和卵巢良性肿瘤间,p27和Cyclin D1的各自表达无明显差异;p27在正常卵巢组织和卵巢良性上皮肿瘤中高表达,在卵巢癌中表达降低(P<0.05),且随着肿瘤分级、分期增高(恶性程度增高),阳性表达率逐渐下降;而Cyclin D1的表达则相反;两者在肿瘤中的表达呈负相关.结论 p27kip表达下降、Cyclin D1过表达可能在卵巢癌的发生中起重要作用,检测p27kip1、Cyclin D1在卵巢癌中的表达可预测肿瘤生物学行为特征,可以作为判断预后的指标.     

卵巢癌中Caspase-3 的表达及与化疗耐药相关性研究

目的:检测Caspase-3在卵巢癌中的表达并探讨其与卵巢癌化疗耐药相关性。方法:利用组织芯片技术结合免疫组化方法,对176例卵巢癌、70例良性卵巢肿瘤、50例正常卵巢组织中Caspase-3的表达进行检测,分析其表达与临床病理特征及化疗疗效的相关性。结果:卵巢癌中Caspase-3的表达显著低于正常卵巢及卵巢良性肿瘤,且卵巢癌FIGO临床分期越晚其表达越低(P<0.01),而与肿瘤的组织类型、病理分级及患者的年龄和月经状况无显著相关性(P>0.05)。Caspase-3在卵巢癌化疗耐药组的阳性率显著低于化疗敏感组(P<0.01)。结论:凋亡蛋白酶Caspase-3的缺表达可能参与了卵巢癌的发生发展过程,在卵巢癌化疗耐药的行程中发挥作用;Caspase-3的检测可能对指导卵巢癌临床化疗用药有帮助,可提高化疗效果和减少化疗耐药,并成为预测化疗疗效的有用指标。     

雌激素受体GPR30在上皮性卵巢癌中的表达及其与MMP-9表达的相关性

本文旨在研究新型雌激素受体——G蛋白耦联受体30(G protein-coupled receptor30,GPR30)在上皮性卵巢癌中的表达,及其与基质金属蛋白酶9(matrix metalloproteinases-9,MMP-9)表达的相关性。收集39例上皮性卵巢肿瘤组织标本,包括上皮性卵巢癌30例(浆液性囊腺癌19例、粘液性囊腺癌5例和子宫内膜样癌6例)、良性上皮性卵巢肿瘤9例,并取4例正常卵巢组织进行对照,应用免疫组织化学法检测GPR30和MMP-9的表达,使用?2检验、Fisher’s精确检验和Spearman等级相关分析等统计学方法对所得数据进行统计分析。结果显示,上皮性卵巢癌中GPR30的高表达率显著高于良性卵巢肿瘤(P0.01)和正常卵巢组织(P0.05);上皮性卵巢癌中MMP-9的高表达率显著高于正常卵巢组织(P0.05),但与良性卵巢肿瘤无明显差异。肿瘤病理类型、分期和患者年龄因素与GPR30高表达率的相关性分析显示:不同病理类型上皮性卵巢癌的GPR30高表达率之间存在显著性差异(P0.05),而各病理类型的MMP-9高表达率之间没有差异;临床III~IV期上皮性卵巢癌的GPR30高表达率显著高于I~II期(P0.05),二者MMP-9高表达率之间没有差异;三个年龄组病例中的GPR30高表达率之间未见明显差异,三者MMP-9高表达率之间亦没有差异。Spearman等级相关分析显示上皮性卵巢癌组织中GPR30与MMP-9的表达之间具有正向相关性(P0.01)。以上结果提示,GPR30可能参与上皮性卵巢癌的发生发展,并与其侵袭和转移具有一定的相关性,可作为卵巢癌早期诊断和恶性程度判断的指标之一。     

PDCD4和DNMT1在卵巢上皮性癌中的表达及其临床意义

目的:检测PDCD4和DNMT1在正常卵巢组织、卵巢良性肿瘤组织、卵巢上皮性癌组织中的表达,并探讨其临床意义。方法:采用免疫组化法检测20例正常卵巢、25例卵巢良性肿瘤、40例卵巢上皮性癌组织中PDCD4和DNMT1的表达情况,分析其与卵巢上皮性癌临床病理参数之间的关系。结果:正常及良性卵巢组织中PDCD4的阳性表达率明显高于卵巢癌组(P0.05),卵巢癌的FIGO分期越高,PDCD4的表达越低,卵巢癌的病理分化程度越低,PDCD4的表达也越低,PDCD4的表达与卵巢癌的组织类型、腹水、年龄、是否绝经无关。正常及良性卵巢组织中DNMT1的阳性表达率明显低于卵巢癌组(P0.05),卵巢癌的病理分化程度越低,DNMT1的表达越高,但其与卵巢癌的FIGO分期、组织类型、腹水、年龄、是否绝经均无关。卵巢癌中DNMT1的与PDCD4的表达呈显著负相关(P0.05)。结论:PDCD4和DNMT1在卵巢癌中的表达呈负相关,PDCD4的表达下调和DNMT1的表达上调可能在卵巢癌的发生及发展中起重要作用。     

雌激素受体、环氧化酶-2在卵巢癌患者中的表达及临床意义

摘要 目的：探讨雌激素受体（ER）、环氧化酶-2（COX-2）在卵巢癌患者中的表达及临床意义。方法：选取经病理手术确诊的120例患者的石蜡卵巢组织切片标本,其中卵巢癌54例,卵巢良性肿瘤66例。采用免疫组化检测ER、COX-2的表达情况。结果：ER和COX-2在卵巢癌组织中的阳性表达率明显高于良性卵巢肿瘤组织（P＜0.05）。Ⅳ期卵巢癌组织中ER阳性表达率明显高于Ⅰ期（P＜0.05）,其余病理分期比较无差异（P>0.05）;不同病理分期的COX-2阳性表达率比较无差异（P>0.05）。不同组织学分级卵巢癌组织中ER阳性表达率组组比较均无差异（P>0.05）。Ⅱ级卵巢癌组织中COX-2阳性表达率明显高于Ⅰ级（P<0.05）,其余病理分级比较无统计学意义（P>0.05）。伴有转移卵巢癌组织中ER阳性表达率明显高于无转移（P<0.05）;伴有转移卵巢癌组织中COX-2阳性表达率与无转移比较无统计学意义（P>0.05）;伴有转移卵巢癌组织中ER、COX-2表达免疫组织化学评分法（IHS）评分均高于无转移（P＜0.05）。结论：ER和COX-2在卵巢癌组织和卵巢良性肿瘤组织中的表达逐渐上调,且在卵巢癌组织的表达高于卵巢良性肿瘤组织;ER的表达与卵巢癌病理分期和转移相关,COX-2与组织学分级相关,二者对卵巢癌病情的预测具有互补性,可作为生物学指标,对卵巢癌恶性程度进行判断评估。     

Nucleostemin基因在卵巢上皮性肿瘤的表达及与病理分型的关系

目的：研究核干细胞因子Nucleostemin（NS）基因在卵巢上皮性肿瘤中的表达,探讨其与肿瘤病理分型的关系。方法：采用RT-PCR及Western blot检测36例卵巢癌组织手术标本,32例卵巢良性上皮肿瘤组织手术标本,12例正常卵巢组织标本中Nu-cleostemin基因及相应蛋白的表达,采用分组对照的方法对比3组样本中NS基因及蛋白的表达情况,并进行相对定量研究。采用统计学方法检测NS基因的表达是否与临床病理分级及血清CA125存在关联。结果：①卵巢癌组织中NS的阳性表达率显著高于良性肿瘤组织及正常卵巢组织;②卵巢癌组织中,淋巴结转移组NS的表达水平高于未转移组;③临床分期Ⅲ期组的表达水平高于ⅠB期组;④中、低分化组的表达水平高于高分化组。结论：卵巢癌组织中存在NS基因的高表达,其表达量与组织类型无关,而与临床TNM分期及组织分级正相关。     

Synthesis and physicochemical properties of 2'-deoxy-2',2'-difluoro-beta-D-ribofuranosyl and 2'-deoxy-2',2'-difluoro-alpha-D-ribofuranosyl oligonucleotides

We present procedures for nucleoside and oligonucleotide synthesis, binding affinity (Tm) and structural analysis (CD spectra) of 2'-deoxy-2',2'-difluoro-alpha-D-ribofuranosyl and 2'-deoxy-2',2'-difluoro-beta-D-ribofuranosyl oligothymidylates. Possible reasons for the thermal instability of duplexes formed between these compounds and RNA or DNA targets are discussed.     

Synthesis and properties of 2'-O-methyl-2-thiouridine and oligoribonucleotides containing 2'-O-methyl-2-thiouridine

A new method for the synthesis of 2'-O-methyl-2-thiouridine (s2Um) found in thermophilic bacterial tRNA was developed. Structural properties of s2Um and s2Um(p)U were studied by using 1H NMR spectroscopy. A modified nonaribonucleotide (RNA*: 5'-CGUUs2UmUUGC-3') was synthesized to study the base-recognition ability of s2Um in formation of RNA-RNA and RNA DNA duplexes. The UV melting experiments revealed that RNA*-RNA and RNA*-DNA duplexes having an s2U-A base pair are more stable than those having a U-A base pair. On the contrary, the thermal stability of RNA*-RNA and RNA*-DNA duplexes having an s2U-G wobble base pair was much lower than that of the unmodified duplexes having a natural U-G base pair. It is concluded that s2Um has higher selectivity toward A over G than unmodified U.     

Stereospecific 2'-amination and 2'-chlorination of adenosine by Actinomadura in the biosynthesis of 2'-amino-2'-deoxyadenosine and 2'-chloro-2'-deoxycoformycin

2'-Amino-2'-deoxyadenosine and 2'-chloro-2'-deoxycoformycin (2'-CldCF) are two nucleoside antibiotics produced by Actinomadura. The biosynthesis of these two nucleoside antibiotics has been studied by the addition of [U-14C]adenosine with or without unlabeled adenine to cultures of Actinomadura. By this experimental approach, it is possible to demonstrate that adenosine is the direct precursor for the biosynthesis of 2'-amino-2'-deoxyadenosine and 2'-CldCF. These conclusions are based on the observation that the percentage distribution of 14C in the aglyconic and pentofuranosyl moieties of 2'-amino-2'-deoxyadenosine and 2'-CldCF were similar to the distribution of 14C in the adenine and ribosyl moieties of the [U-14C]adenosine (i.e., 48:52) added to cultures of Actinomadura. Experimentally, the percentage distribution of 14C in the (i) adenine:2-amino-2-deoxy-beta-D-ribofuranose of 2'-amino-2'-deoxyadenosine is 51:49; (ii) 8-(R)-3,6,7,8-tetrahydroimidazo[4,5-d]-[1,3-diazepin-8-o1]:2 -chloro-2- beta-D-ribofuranose of 2'-CldCF is 45:55; and (iii) adenine:ribose of the adenosine isolated from the RNA of Actinomadura is 42:58. Further proof that adenosine is the direct precursor for the biosynthesis 2'-amino-2'-deoxyadenosine and 2'-CldCF was demonstrated by the addition of 75 mumol of unlabeled adenine together with [U-14C]adenosine to nucleoside-producing cultures of Actinomadura. The percentage distribution of 14C in the aglycon and the sugar moieties of 2'-amino-2'-deoxyadenosine and 2'-CldCF were 46:54 and 47:53, respectively; the percentage distribution of 14C in the adenine and ribose moieties of the adenosine isolated from the RNA of Actinomadura was 51:49. These data show that the hydroxyl on C-2' of the ribosyl moiety of adenosine undergoes a replacement by a 2'-amino or a 2'-chloro group to form 2'-amino-2'-deoxyadenosine or 2'-CldCF with retention of stereconfiguration at C-2'. Finally, Actinomadura can utilize inorganic chloride from the medium as demonstrated by the isolation of [36Cl]2'-CldCF following the addition of [36Cl]chloride to the culture medium. Mechanisms for the regioselective modification of the C-2' hydroxyl group and stereospecific insertion of the amino and chloro groups are discussed.     

Stereoselective Synthesis of 2-Amino-2-deoxy-d-arabinose and 2-Deoxy-d-ribose

An efficient method for the stereoselective synthesis of 2-amino-2-deoxy-d-arabinose and 2-deoxy-d-ribose is described.

The key step in this method was accomplished by the nucleophilic addition of methyl isocyanoacetate to 2,3-O-isopropylidene-d-glyceraldehyde with high erythro-selectivity (nearly 100%).

Subsequent intermolecular cyclization predominantly gave the desired oxazoline derivative (trans-form), in which two new chiral centers were formed. The oxazoline derivative was efficiently converted to both 2-amino-2-deoxy-d-arabinose and 2-deoxy-d-ribose.     

Parasiticidal 2-alkoxy- and 2-aryloxyiminoalkyl trifluoromethanesulfonanilides

A series of novel 2-alkoxy- and 2-aryloxyiminoalkyl trifluoromethanesulfonanilide derivatives have shown significant in vitro parasiticidal activity against the ectoparasites Ctenocephalides felis and Rhipicephalus sanguineus. A number of these compounds also displayed significant in vitro endoparasite activity against the nematode Haemonchus contortus.