端粒酶在妇科肿瘤诊治方面的应用进展

不同的妇科肿瘤细胞都有端粒酶活性，而大多数正常体细胞却未见表达。端粒酶可能成为妇科肿瘤诊断和估预后的异特标记物。端粒酶抑制剂可能用于治疗妇科肿瘤。     

卵巢肿瘤组织中端粒酶活性测定及其临床意义的研究

目的：研究测定端粒酶活性在诊断和预测卵巢肿瘤患者预后方面的意义。方法：应用改良的非放射性同位素端粒重复序列扩增实验记录（ＮＲＩＴＲＳＡＰ）检测２６份卵巢肿瘤组织（卵巢良性肿瘤６份,卵巢交界性或低度恶性肿瘤５份,卵巢恶性肿瘤１５份）的端粒酶活性。结果：６份卵巢良性肿瘤组织中均未测出端粒酶活性。５份卵巢交界性或低度恶性肿瘤组织中３份（６０％）端粒酶活性阳性表达;１５份卵巢恶性肿瘤组织中１４份（９３３％）端粒酶活性阳性表达。卵巢恶性肿瘤组织中,端粒酶活性阳性率均高于卵巢交界性肿瘤及卵巢良性肿瘤组织中者（Ｐ＜００５）;卵巢交界性肿瘤组织中者高于卵巢良性肿瘤组织中者（Ｐ＜００５）,卵巢恶性肿瘤组织中强阳性率（）及（）高于卵巢交界性肿瘤组织中者（Ｐ＜００５）。在卵巢上皮性恶性肿瘤组织中,在组织类型、组织学分级及肿瘤分期之间差异无显著性,但总的端粒酶活性阳性率,浆液性癌中（）及（）表达者高于其他类型,低分化者高于高分化者,晚期者高于早期者。结论：端粒酶活性的检测不仅有助于卵巢肿瘤的诊断和鉴别诊断,而且有助于预测患者的预后。     

子宫颈癌和正常宫颈组织雌,孕激素受体的研究

采用单克隆酶联免疫测定法，检测了２１例宫颈癌（宫颈癌组）和１７例正常宫颈肌层纤维组织（正常组）的雌、孕激素受体水平。结果：正常组雌激素受体（ＥＲ）的阳性率为８８．２％，孕激素受体（ＰＲ）的阳性率为７４．５％，ＥＲ和ＰＲ均阳性者占７４．５％。宫颈癌组ＥＲ阳性率为６６．７％，ＰＲ阳性率为４２．９％，ＥＲ和ＰＲ均阳性者占３８．１％。两组ＥＲ状态和水平比较，差异无显著意义（Ｐ＞０．０５），但宫颈癌组的Ｐｋ阳性率和ＰＲ水平均明显低于正常组（Ｐ＜０．０５）。宫颈癌组织的ＥＲ和ＰＲ水平与肿瘤分期，组织学分级之间，差异无显著意义（Ｐ＞０．０５）。绝经前宫颈癌组织ＰＲ水平较绝经后明显升高。宫颈腺癌组织ＥＲ和ＰＲ水平均明显高于宫颈鳞癌。宫颈癌患者按血清雌二醇水平高低分组，ＰＲ水平和ＰＲ／ＥＲ比值比较，差异均有显著意义。     

检测妊娠区带蛋白临床意义的研究

采用血清妊娠区带蛋白（ＰＺＰ）单向免疫扩散法时７０８例正常妊娠孕妇，２０７例异常妊娠孕妇及１８８例妇科肿瘤患者进行测定。结果表明：正常妊娠孕妇血清ＰＺＰ含量在妊娠第５周即可测出，其含量随孕周的增加而增加，至妊娠４０周达高峰；先兆流产预后佳者，８１．５％分布在正常范围内；妊高征、胎儿宫内生长迟缓、无脑儿、异位妊娠时，ＰＺＰ多在正常范围内；５１．６％恶性葡萄胎患者血清ＰＺＰ含量低于正常范围，绒毛膜癌低值者占８０．０％，在妇科肿瘤中，卵巢癌患者血清ＰＺＰ明显高于卵巢瘤（Ｐ＜０．０１），子宫内膜癌和宫颈癌高于子宫肌瘤（Ｐ＜０．０１）。本研究认为，测定血清ＰＺＰ含量可作为判定先兆流产预后，鉴别滋养细胞肿瘤及妇科良恶性肿瘤的一项重要参考指标。     

卵巢肿瘤组织中多药耐药基因的表达及其临床意义

目的：检测卵果肿瘤组织中多药耐药基因的表达并探讨其临床意义。方法；采用ＲＮＡ多聚酶链反应（ＲＴ－ＰＣＲ）方法，对３５例卵巢恶性肿瘤，２０例卵巢良性肿瘤和１７例正常卵巢组织标本进行我药耐药基因＊（ＭＤＲ１）表达的测定。结果：（１）卵巢恶怀肿瘤组织中ＭＤＲ１表达阳性率为５７．１４％，而卵巢良性肿瘤和正常卵巢组织中则无一例ＭＤＲ１表达阳性。（２）卵巢恶性肿瘤中，初治者和复发者的ＭＤＲ１表达阳性率分别为４     

妇科肿瘤组织中线粒体DNA基因突变的研究

目的 探讨原发性肿瘤组织中线粒体DNA(mtDNA)基因突变及其在肿瘤发生、发展中的作用。方法 采用聚合酶链反应-单链构象多态性分析(PCR—SSCP)及DNA测序方法,对32例妇科恶性肿瘤患者(观察组)、8例妇科良性肿瘤患者(对照组)的肿瘤组织和瘤旁正常组织标本进行mtDNA基因突变检测。观察组中子宫颈癌5例,子宫内膜癌10例,卵巢癌17例。对照组中,卵巢上皮性肿瘤4例,子宫肌瘤4例。结果 观察组mtDNA基因突变率和多态性频率分别为68．8％和56．3％。对照组mtDNA基因突变率和多态性频率分别为2／8和4／8。两组mtDNA基因突变率比较,差异有显著性(P＜0．05);mtDNA基因突变热点位于Cytb基因区;mtDNA基因突变为多基因、多位点的突变,其中63．6％涉及到2个以上的基因。结论 在妇科恶性肿瘤组织中,存在mtDNA基因编码区的高频率变异,这种变异可能是妇科恶性肿瘤发生机理中的一个重要的核外分子遗传学改变。     

人宫颈癌脱落细胞端粒酶活性检测的意义

目的 研究宫颈上皮内瘤样病变(CIN )和宫颈癌脱落细胞端粒酶激活的意义。方法 采用端粒酶PCR-ELISA法检测13例CIN和17例宫颈癌脱落细胞端粒酶活性,并与相应的组织标本中端粒酶活性相比较,11例正常宫颈脱落细胞作为对照。结果8例(61.54％)CIN、14例(82.35％)宫颈癌脱落细胞表达端粒酶活性,正常宫颈无端粒酶活性,CIN、宫颈癌及正常宫颈脱落细胞端粒酶活性平均值分别为0.328±0.192、1.329±0.259和0.039±0.084,三组间端粒酶表达率和量存在显著差异。CINⅠ、Ⅱ、Ⅲ级之间端粒酶活性无明显差异。CIN及宫颈癌组织中端粒酶活性和相应脱落细胞一致。结论 端粒酶的激活是宫颈癌的早期改变,在宫颈癌发生发展过程中起重要作用;宫颈脱落细胞和组织中端粒酶活性可能成为宫颈癌早期诊断、预后判断和肿瘤浸润的标记物。     

端粒酶激活与上皮性卵巢癌化疗敏感性及其预后的关系

目的：探讨端粒酶活性水平对卵巢化疗疗效及预后影响。方法：以TRAP－PCR－ELISA法检测卵巢癌组织中端粒酶活性;以MTT法检测卵巢癌细胞对CDDP、ADM敏感。结果：22例卵巢癌组织中,端粒酶阴性者3例（13．64％）,阳性者19例（86．36％）。8例端粒酶活性高于2U者,PAC方案化疗有效者5例（62．5％）,11例端粒酶活性低于2U者,化疗均有效（100％）。端粒酶活性高于2U者,进展与复发或死亡5例（62．5％）,而端粒酶活性低于2U者仅1例出现复发,无死亡。结论：随着端粒酶活性增高,卵巢癌对CDDP、ADM敏感性下降。     

端粒、端粒酶与妇科恶性肿瘤早期诊断及肿瘤恶变预测

端粒是染色体末端独特的蛋白质－DNA结构，具有保护染色体作用。端粒酶是由RNA和蛋白质亚基组成的一种特殊反转录酶，能延长端粒。对许多系统内端粒酶的检测结果表明：在肿瘤组织内通常表达，而在对照正常组织内几乎不表达。端粒酶活性与肿瘤发生发展密切相关，可作为肿瘤的诊断和判断预后的指标。     

端粒、端粒酶与妇科恶性肿瘤早期诊断及肿瘤恶变预测

端粒是染色体末端独特的蛋白质—DNA结构，具有保护染色体作用。端粒酶是由RNA和蛋白质亚基组成的一种特殊反转录酶，能延长端粒。对许多系统内端粒酶的检测结果表明：在肿瘤组织内通常表达’而在对照正常组织内几乎不表达。端粒酶活性与肿瘤发生发展密切相关，可作为肿瘤的诊断和判断预后的指标。     

Quantitative telomerase activity in malignant, benign and normal gynecological tissues

OBJECTIVE: The aim of this study was to evaluate quantitative telomerase activity in malignant, benign and normal gynecological tissue samples by using the Telomerase-PCR ELISA kit, and to determine a cut-off level for malignancy by this quantitative method. MATERIALS AND METHODS: Fifty gynecological tumors, 27 benign gynecological disorders and 29 normal tissues were analyzed by the Telomerase-PCR ELISA kit. All tissues were confirmed by a pathologist. A ROC (receiver operator characteristic) curve was drawn to determine a threshold level best discriminating malignant tissues from benign pathologies and normal tissues. Telomerase activity was compared in malignant, benign and normal tissues. RESULTS: The mean level of telomerase activity of the malignant tumor samples (1.03 +/- 0.53 units) was significantly (p < .001) higher than the normal tissues (0.13 +/- 0.07 units) and benign pathologies (0.37 +/- 0.25 units). The cut-off point to differentiate malignant samples from benign samples was set at 0.42 units, where the sensitivity was 93.8% and the specificity was 89.3%. Positive predictive value was 84% and negative predictive value was 89.3%. There was a significant difference in telomerase activity between malignant, benign and normal tissues within each histological group. CONCLUSION: In this preliminary study, the telomerase-PCR ELISA method was found to have a high sensitivity and specificity to differentiate malignant gynecological tissues from benign tissues.     

Diagnostic value of telomerase activity in gynecologic malignancies

Nagai N, Murakami J, Oshita T, Ohama K, Tahara H. Diagnostic value of telomerase activity in gynecologic malignancies. Int J Gynecol Cancer 1998; 8 : 481–488.
We investigated the diagnostic significance of telomerase activity in gynecological malignancies. Tissue samples were obtained from 24 cervical cancers, 27 uterine cancers (22 endometrial cancers and five sarcomas), 33 ovarian cancers (31 epithelial tumors and 2 germ cell tumors), and 11 benign ovarian tumors. In addition, cervical cytology specimens were obtained from 30 squamous intraepithelial lesions (13 low grade and 17 high grade), and from 22 normal females. Telomerase activity was detected using the TRAP assay, and the relative telomerase activity was obtained using the BioMax DNA image analysis system. Telomerase activity was detected in 22/24 (91.7 %) cervical cancers, 23/27 (85.2%) uterine tumors and 30/33 (90.9%) ovarian cancers. Weak telomerase activity was detected in two mature cystic teratomas and also found in 9/17 (52.9%) high grade SIL and 2/13 (15.4%) low grade lesions. Telomerase activity showed no relationship with tumor histology or clinical stage, and there was no statistically significant difference between patients with uterine cancer and ovarian cancer. Relative telomerase activity showed a correlation with the dilution assay, and significantly higher telomerase activity was found in uterine cervical cancer compared with precancerous lesions and in ovarian cancer compared with benign ovarian tumors. After establishment of an assay for telomerase, it may be useful for cancer diagnosis and identification of high-risk groups.     

Analysis of Telomerase Activity in Ovarian Cystadenomas, Low-Malignant-Potential Tumors, and Invasive Carcinomas

OBJECTIVE: Inappropriate telomerase expression has been reported to be associated with the development and/or progression of malignancies. Therefore, the purpose of the study was to determine and evaluate the levels of telomerase activity in normal ovary, cystadenomas, low-malignant-potential tumors, and carcinomas of the ovary. METHODS: In the present study, telomerase activity was examined in frozen tissue specimens of normal ovary (n = 6), ovarian cystadenomas (n = 13), ovarian low-malignant-potential (LMP) tumors (n = 12), and ovarian invasive carcinomas (n = 81). Clinicopathological information including age at diagnosis, histological grade, FIGO stage, presence of distant metastasis at diagnosis, and residual disease was available for all patients with ovarian carcinomas (n = 81). Telomerase activity was assessed by the telomeric repeat amplification protocol (TRAP). Arbitrary values were assigned to processivity of telomerase activities based on the molecular weights of the telomeric repeat DNA ladders, and were graded as "negative," "moderate" (< or =99 bp), or "high" (>100 bp) activities. The specificity of telomerase activity was determined by the pretreatment of telomerase-positive control or tumor samples with RNase that led to the abolition of the activity. In addition, to determine the possibility of false negativity due to the presence of telomerase inhibitors, TRAP assay was performed on each of the telomerase-negative specimens by mixing them individually with the telomerase-positive control. RESULTS: Telomerase activity in the progression of ovarian carcinogenesis was evaluated. In comparison with normal ovary/cystadenoma (32%), a much higher frequency of the moderate activity was observed in LMP tumors (67%) or invasive carcinomas (57%), suggesting a close association between the latter two categories. The results reflect a subpopulation of telomerase-positive LMP tumor cells with the potential to develop invasive carcinomas. None of the specimens of the benign or LMP tumors exhibited high activity. In contrast, 18% of ovarian invasive carcinomas showed high telomerase activity (P = 0.013, Fisher exact test) and further 57%, moderate activity (75% in all). A statistically significant difference was observed in the expression of telomerase activity between normal ovary/benign cystadenomas and ovarian invasive carcinomas (P = 0.001, chi(2) test). CONCLUSIONS: The study showed a high prevalence of telomerase activity in LMP tumors or invasive carcinomas, the high levels of telomerase activity being associated exclusively with the invasive ovarian carcinomas. Therefore, the levels of processivity of telomerase activity and evidence of its statistically significant association with ovarian carcinoma suggest its role in the progression of ovarian carcinogenesis.     

CyclinD1、C-erbB2及bcl-2基因在卵巢上皮性肿瘤中的表达及其临床意义

目的 :检测癌基因CyclinD1、C erbB2及bcl 2在卵巢上皮肿瘤中的表达 ,探讨它们在卵巢肿瘤发生、发展中的作用及临床、病理意义。方法 :应用免疫组化SP法检测72例恶性卵巢肿瘤、12例交界性卵巢肿瘤、2 1例良性肿瘤及 10例正常卵巢组织中Cy clinD1、C erbB2及bcl 2基因的表达情况。结果 :1.卵巢恶性、交界性及良性肿瘤中Cy clinD1阳性率依次为 2 7.78%、33.3%、9.5 2 %。恶性及交界性肿瘤阳性率明显高于良性肿瘤 ,其阳性率与组织学分级呈负相关 ,而与患者年龄、临床分期、组织学类型无关 ;2 .卵巢恶性、交界性及良性肿瘤中C erbB2的阳性率依次为 5 6 .9%、4 1.6 7%、14.2 8%。恶性及交界性肿瘤阳性率明显高于良性肿瘤 ,差异有显著性。C erbB2阳性表达在组织分化差及期别晚的肿瘤中较分化好、期别早者高 ;3.卵巢恶性、交界性、良性肿瘤中bcl 2的阳性表达率依次为 6 3.89%、5 0 %、2 8.5 %。恶性及交界性肿瘤与良性肿瘤之间的表达差异有显著性。组织分化差、期别早的肿瘤中bcl 2的阳性率较分化好、期别晚者高 ;4 .两种及两种以上基因同时表达率 (5 1.4 % )显著高于单基因表达 (2 7.79% )。CyclinD1与C erbB2基因表达呈负相关。结论 :CyclinD1、C erbB2及bcl 2基因在卵巢癌发生、发展中起重要作用 ,表明细胞?     

Selenium concentrations in serum, hair and tumor tissue from patients with ovarian tumors

Selenium (Se) concentration was determined in 150 samples of serum, 135 samples of hair and 96 samples of ovarian tissue from patients with malignant or benign ovarian tumors and normal subjects. The serum Se level in patients with malignant ovarian tumors was significantly lower than that in patients with benign ovarian tumors and normal subjects. No difference of Se concentrations of hair among different groups was observed. Se level in malignant ovarian tumor tissue was significantly higher than that in benign ovarian tumor tissue and normal ovarian tissue. The low serum Se level in patients with malignant ovarian tumors was probably the result of protective migration of Se from blood to the cancer tissue.     

Quantitative biochemical analysis of lactate dehydrogenase in human ovarian tissues: correlation with tumor grade

Abstract.   Šimaga Š, Osmak M, Babič D, Šprem M, Vukelič B, Abramič M. Quantitative biochemical analysis of lactate dehydrogenase in human ovarian tissues: correlation with tumor grade. Int J Gynecol Cancer 2005; 15: 438–444.
In an attempt to identify glycolytic capacity of normal and neoplastic human ovary, total lactate dehydrogenase (LDH) activity was measured in tissue cytosol originating from 69 patients (18 with benign ovarian tumor, 34 with ovarian carcinoma, six with nonepithelial ovarian malignant tumors, and 11 with tumor metastatic to ovary) and compared to the LDH activity of normal ovarian tissues ( n = 19). Median value of total LDH-specific activity expressed as U/mg protein was 0.546 in normal tissues, 0.584 in benign tumors, 1.071 in malignancies metastatic to ovaries, 0.872 in nonepithelial primary ovarian tumors, and 0.818 in primary carcinomas. A significant rise in LDH-specific activity was found in malignant primary and secondary tumors of epithelial and nonepithelial origin, but not in benign neoplasms, compared to the activity in normal tissue. Ovarian carcinomas of serous histologic type did not differ in LDH activity from mucinous tumors. However, poorly differentiated carcinomas (grade 3) showed significantly enhanced activity of this glycolytic enzyme when compared to its grade 1 counterpart. The subgroup of grade 1 tumors did not differ in LDH activity from normal and benign ovarian tissue. Obtained results suggest that direct correlation might exist between ovarian epithelial tumor grade and lactate dehydrogenase activity.