类固醇硫酸酯酶与乳腺癌

类固醇硫酸酯酶(STS)在乳腺癌组织中表达水平明显高于非恶性组织,并与乳腺癌患者的临床病理指标及预后相关.乳腺癌组织通过STS可以合成具有生物活性的雌激素和具有雌激素样作用的甾体类化合物,促进癌细胞增殖.抑制STS的活性可以减缓肿瘤细胞的生长,目前已有部分STS抑制剂在体内外试验中显示了对肿瘤细胞较好的抑制作用.     

STS和AROM在乳腺癌组织中的表达及其临床意义

目的 探讨类固醇硫酸酯酶（STS）和芳香化酶（AROM）在乳腺癌组织中的表达情况及其与临床病理特征的关系。方法 应用免疫组化法检测82例乳腺癌组织及癌旁组织中STS和AROM的表达情况。结果 STS在乳腺癌和癌旁组织中的阳性表达率分别为73.1％（60／82）和39.0％（32／82）,差异具有统计学意义（P=0.000）。AROM在乳腺癌和癌旁组织中的阳性表达率分别为58.5％（48／82）和39.0％（32／82）,差异具有统计学意义（P=0.012）。在乳腺癌组织中,STS表达与HER 2表达呈正相关（r=0.269,P=0.015）,与其他临床病理特征无关;AROM表达与淋巴结转移相关（r=0.268,P=0.015）,与其他临床病理特征无关;STS与AROM表达之间无相关性（r=-0.040,P=0.720）。结论 STS和AROM表达均与乳腺癌的发生、发展密切相关,其中STS途径比AROM途径可能更重要。     

乳腺癌组织中环氧化酶-2的表达及其意义

目的：观察环氧化酶-2(cyclooxygenase-2，COX-2)在乳腺癌中的表达，并探讨其临床病理意义。方法：采用免疫组织化学方法检测了80例乳腺癌、27例乳腺不典型增生以及20例正常乳腺组织中COX-2表达并分析其与临床病理的相关性。结果乳腺癌组织中C0X-2阳性表达率为57．5％(46／80)，明显高于乳腺不典型增生11．1％(3／27)和正常乳腺组织10．O％(2／20)，P＜O．01。COX-2表达与淋巴结转移、临床分期组织学分级相关，P＜O．05。COX-2高表达的乳腺癌术后5年无病生存率明显高，于COX-2低表达者，P＜O．05。结论：COX-2表达可以作为判定乳腺癌恶性程度和预后的确效生物学指标。     

乳腺癌组织中环氧化酶-2的表达及其意义

目的 :观察环氧化酶 -2 (cyclooxy genase 2 ,COX 2 )在乳腺癌中的表达 ,并探讨其临床病理意义。方法 :采用免疫组织化学方法检测了 80例乳腺癌、2 7例乳腺不典型增生以及 2 0例正常乳腺组织中COX 2表达并分析其与临床病理的相关性。结果 :乳腺癌组织中COX 2阳性表达率为 5 7 5 % ( 4 6/80 ) ,明显高于乳腺不典型增生 11 1% ( 3 / 2 7)和正常乳腺组织 10 0 % ( 2 / 2 0 ) ,P <0 0 1。COX 2表达与淋巴结转移、临床分期、组织学分级相关 ,P <0 0 5。COX 2高表达的乳腺癌术后 5年无病生存率明显高于COX 2低表达者 ,P <0 0 5。结论 :COX 2表达可以作为判定乳腺癌恶性程度和预后的有效生物学指标     

STS和EST在乳腺癌患者中的表达及临床意义

目的 探讨硫酸酯酶(steroid sulfatase,STS)、雌激素硫酸转移酶(estrogen sulfotransferase,EST)在乳腺癌组织和正常乳腺组织中的表达与临床病理特征的关系,及STS与EST相互之间的关系.方法 选取术前未行放疗、化疗及内分泌治疗的乳腺癌患者82例,取其术后肿瘤组织和距肿瘤边缘＞5 cm的乳腺组织.应用免疫组织化学法,检测STS、EST在乳腺癌组织及正常乳腺组织中的表达,采用卡方检验和spearman法分析其表达及相关性.结果 乳腺癌组织中STS蛋白表达水平高于正常乳腺组织,差异有统计学意义(P=0.000).EST在乳腺癌和正常乳腺组织中的表达差异无统计学意义(P =0.367).在乳腺癌组织中STS与EST的表达水平无差异,两者无相关性(r =0.078,P=0.487).结论 STS、EST的表达与乳腺癌的发生、发展相关.     

增殖抑制基因在乳腺癌组织中的表达及其意义

目的：探讨新的增殖抑制基因（hypevplasia suppressor gene，HSG）在乳腺癌组织中的表达及其意义。方法：应用免疫组织化学和原位杂交法检测55例乳腺癌标本和18例癌旁正常乳腺组织标本中HSG的表达。应用Image Proplus数码图像分析系统测定组织切片的积分吸光度（D）值，分析其与临床病理参数的相关性。结果：HSGmRNA及其蛋白的阳性染色主要集中于细胞质，呈棕黄色颗粒。55例乳腺癌标本中HSG mRNA及其蛋白的阳性表达率分别为40．00％（22／55）和41．82％（23／55），而癌旁正常乳腺组织的表达率分别为83．33％（15／18）和88．89％（16／18），2组比较，差异有统计学意义（P〈0．05）。在乳腺癌组织中HSGmRNA及其蛋白的低表达与组织学分级、淋巴结转移和雌激素受体（estrogen receptor，ER）有关（P〈0．05），与肿瘤大小和年龄等无关（P〉0．05）。结论：HSG在不同乳腺组织中的表达量不同，提示HSG可能与乳腺癌的发生、发展有密切关系。     

雌激素调节蛋白PS2在乳腺癌组织中的表达及其应用

目的 评价雌激素调节蛋白PS2在乳腺癌组织中的表达以及在各种乳腺癌中的分布以及在雌、孕激素受体的相互关系的价值。方法 应用放免法检测68例原发乳腺癌中的PS2含量,同时用DCC方法检测雌、孕激素受体含量。结果 68例乳腺癌组织中的PS2含量从0．6－58ng/mg蛋白,以≥2ng/mg蛋白为阳性,阳性率57．4％（39／68）,雌激素受体,孕激素受体以≥10fmol/mg蛋白为阳性,在50例ER（＋）病人中有30例PS2（＋）（60．0％）,而在ER（＋）PR（＋）22例病人中,PS2阳性率达72．7％,但是ER（－）PR（一）病人中阳性率42．9％,两组相比差异有高度显著性（P＜0．01）,PS2阳性率在乳腺癌病理类型中以及临床TNM分型中未见差异,PS2的阳性率在组织学分级中Ⅰ,Ⅱ级明显高于Ⅲ级有统计学意义。结论 PS2与雌激素受体呈正相关关系,在乳腺癌中PS2水平高则预后好,ER、PR、PS2三项指标全阳性的乳腺癌病人可以作为对内分泌治疗有效的新的亚型。     

热休克蛋白5在乳腺癌组织中的表达及其意义

目的 探讨乳腺癌组织中热休克蛋白5（HSPA5）的表达及其与乳腺癌临床病理因素的关系.方法 对乳腺癌手术切除标本中的癌组织97例以及其中带有癌旁正常组织的标本40例,用免疫组织化学二步法检测HSPA5蛋白表达情况.结果 HSPA5在97例乳腺癌中的阳性表达率为81.4％（79/97）.在所选取的40例乳腺癌及癌旁正常组织中,HSPA5的阳性表达率分别为32.5％（13/40）、7.5％（3/40）,差异有统计学意义（x 2=7.813,P=0.038）.HSPA5在不同组织学类型及不同肿瘤直径乳腺癌中的表达差异均无统计学意义（x2=5.785、1.500,P=0.056、0.296）;而在不同组织学分级和有无淋巴结转移的乳腺癌中,表达差异有统计学意义（x2=22.233、5.342,P=0.007、0.024）.结论 HSPA5的在乳腺癌组织中高表达,可能与乳腺癌的发生、发展和浸润转移有关,对于判断乳腺癌的临床预后有重要意义.     

PTEN蛋白在乳腺癌组织中的表达及其临床意义

目的 探讨抑癌基因PTEN在乳腺癌组织中的表达及其临床意义。方法 采用SABC免疫组织化学方法，检测了62例乳腺癌、15例癌旁及12例正常乳腺组织中PTEN蛋白表达水平，结合临床病理指标进性分析。结果 乳腺癌组PTEN蛋白阳性表达率（56．5％）显著低于癌旁组（86．7％）和正常组（100．0％），P均〈0．05；癌旁组（86．7％）低于正常组（100．0％），P〉0．05。伴有淋巴结转移的乳腺癌中PTEN蛋白阳性表达率（32．1％）显著低于无淋巴结转移乳腺癌（61．8％），P〈0．05；浸润性癌PTEN蛋白阳性表达率（44．4％）显著低于早期浸润性癌（75．0％），P〈0．05；PTEN蛋白表达与肿瘤大小、PTNM分期无关，P〉0．05。结论 PTEN蛋白在乳腺癌的发生、发展、侵袭转移中可能起着重要作用。PTEN蛋白表达水平可以作为判定乳腺癌病理生物学行为的客观指标。     

Ezrin和AKT2在乳腺癌组织中的表达及意义

目的 探讨乳腺癌组织中Ezrin、AKT2的表达及其与临床病理指标的关系。方法 应用免疫组织化学SP法,检测56例乳腺癌组织、30例癌旁正常乳腺组织石蜡切片中Ezrin、AKT2的表达。结果 （1）56例乳腺癌组织中Ezrin、AKT2的表达率分别为62.50%和76.79%,与正常乳腺组织相比差异有统计学意义(P<0.05)。（2）Ezrin的表达与乳腺癌的组织学分级、临床TNM分期和淋巴结转移有关(P<0.05),而与原发肿瘤大小、患者年龄大小、病理类型无关(P>0.05)。AKT2的表达与乳腺癌的淋巴结转移有关(P<0.05),与乳腺癌的原发肿瘤大小、组织学分级、病理类型、临床TNM分期以及患者年龄大小无关(P>0.05)。（3）在乳腺浸润性导管癌中Ezrin与AKT2表达水平呈显著正相关(P<0.05)。结论 Ezrin、AKT2高表达与乳腺癌的发生、发展及浸润转移有关,可作为判断乳腺癌生物学行为和预后的重要指标。     

Significance of steroid sulfatase expression in human breast cancer

The sulfatase pathway has been thought to be a primary means of local production of estrone in human breast cancer tissue. We measured steroid sulfatase (STS) mRNA levels in 97 breast cancers and evaluated its association with disease-free survival. High levels of STS mRNA proved to be a significant predictor of reduced relapse-free survival, both as a continuous variable (log STS mRNA; P=0.028) and as a dichotomous variable with an optimized cutoff point (P=0.002). In multivariate analysis a high level of STS mRNA was an independent factor for predicting relapse-free survival. These results suggest a putative role of STS in breast cancer growth and metastasis, and administration of sulfatase inhibitors to breast cancer patients with high levels of STS mRNA might be an additional treatment option.     

A novel steroidal selective steroid sulfatase inhibitor KW-2581 inhibits sulfated-estrogen dependent growth of breast cancer cells in vitro and in animal models

We screened a series of 17β-(N-alkylcarbamoyl)-estra-1,3,5(10)trine-3-O-sulfamate derivatives, and describe here a potent and selective steroid sulfatase (STS) inhibitor with antitumor effects in breast cancer models in vitro and in vivo. In biochemical assays using crude enzymes isolated from recombinant Chinese hamster ovary cells expressing human arylsulfatses (ARSs), one of the best compounds, KW-2581, inhibited STS activity with an IC₅₀ of 4.0 nM, while > 1000-fold higher concentrations were required to inhibit the other ARSs. The failure to stimulate the growth of MCF-7 human breast cancer cells as well as in uteri in ovariectomized rats indicated the lack of estrogenicity of this compound. In MCF-7 cells transfected with the STS gene, termed MCS-2 cells, KW-2581 inhibited the growth of cells stimulated by estrone sulfate (E1S) but also 5-androstene-3β, 17β-diol 3-sulfate (ADIOLS) and dehydroepiandrostenedione 3-sulfate. We found that oral administration of KW-2581 inhibited both E1S- and ADIOLS-stimulated growth of MCS-2 cells in a mouse hollow fiber model. In a nitrosomethylurea-induced rat mammary tumor model, KW-2581 induced regression of E1S-stimulated tumor growth as effectively as tamoxifen or another STS inhibitor, 667 Coumate. Dose-response studies in the same rat model demonstrated that more than 90% inhibition of STS activity in tumors was necessary to induce tumor shrinkage. STS activity in tumors has well correlated with that in leukocytes, suggesting that STS activity in leukocytes could be used as an easily detectable pharmacodynamic marker. These findings demonstrate that KW-2581 is a candidate for development as a therapeutic agent for the treatment of hormone receptors-positive breast cancer.     

Efficacy of three potent steroid sulfatase inhibitors: pre-clinical investigations for their use in the treatment of hormone-dependent breast cancer

Estrogenic steroids, such as estradiol, are known to play a crucial role in the development and growth of hormone-dependent breast cancer. Steroid sulfatase (STS) inhibitors that can prevent the biosynthesis of these steroids via the sulfatase pathway offer therapeutic potential. We show here the in vivo profile, including the efficacy in a xenograft breast cancer model and pharmacokinetics, of three potent STS inhibitors. MCF-7 cells stably over-expressing STS cDNA (MCF-7STS) were generated. Ovariectomised, MF-1, female nude mice receiving subcutaneous injections of estradiol sulfate (E2S) and bearing MCF-7STS xenografts, were orally treated with the STS inhibitors STX64, STX213, and STX1938. Treatment was administered once weekly at a dose of 1 mg/kg for 35 days during which animals received E2S thrice weekly. Mice were weighed and tumor measurements taken weekly. Furthermore, the pharmacokinetics for STX213 was determined in rats. STX213 and STX1938 exhibited potent STS inhibition in vivo. However, STX1938 demonstrated a greater duration of activity. In vehicle treated nude mice receiving E2S, tumor volumes increased by 260% after 35 days compared to day zero. STX64 (1 mg/kg) failed to reduce tumor growth when given once weekly. STX213 and STX1938 (once weekly, 1 mg/kg) significantly inhibited (P < 0.05) tumor growth over this same time period. These compounds completely inhibited liver and tumor STS activity and significantly reduced the levels of plasma E2. This study indicates that the STS inhibitor, STX213, exhibits excellent efficacy and pharmacokinetics and therefore offers a potentially novel treatment for hormone-dependent breast cancer.     

Inhibition of steroid sulfatase activity and cell proliferation in ZR-75-1 and BT-474 human breast cancer cells by KW-2581 in vitro and in vivo

In the present study, we found that two hormone receptor-positive human breast cancer cell lines, ZR-75-1 and BT-474, naturally expressed steroid sulfatase (STS) protein and had catalytic activity to produce estrone from estrone sulfate (E1S) with a comparable level to those in human breast cancer tissues. E1S at physiological concentrations stimulated the growth of those cells. A novel steroidal STS inhibitor, KW-2581 inhibited the STS activity of ZR-75-1 cells with an IC₅₀ of 13 nM, a potency equal to or higher than that of the non-steroidal STS inhibitor, 667 COUMATE. The inhibitory effect of KW-2581 was enhanced by pre-incubation with STS enzyme, suggests being irreversible inhibition. KW-2581 inhibited the E1S-stimulated growth of ZR-75-1 cells with an IC₅₀ of 0.18 nM, but failed to inhibit the growth stimulated by 17β-estradiol. Expression of E1S-induced progesterone receptors in ZR-75-1 cells was reduced by treatment of KW-2581 at concentrations as low as 0.1 nM. Oral administration of KW-2581 for 4 weeks caused tumor shrinkage in a mouse xenograft model. Tumor STS activity had been completely (>95%) eliminated by 24 hours after the last administration. These findings suggest that KW-2581 has considerable potential for therapeutic development as a novel anti-hormonal drug for treatment of breast cancer.     

Localization of estrone sulfatase in human breast carcinomas

We generated anti-human El-STS monoclonal antibodies to localize estrone sulfatase (E1-STS) in human breast carcinomas. In particular, we examined the MCF-7 clone E3, ZR-75-1, MDA-MB 231, and MDA-MB-468 breast cancer cell lines and 25 breast carcinomas by either immunohistochemistry or Western blotting analysis. Simultaneously, we analyzed histological data, estrogen receptor (ER) status, progesterone receptor (PgR) status and epidermal growth factor receptor (EGFR) in breast tissue. All were surgical specimens from female patients. Nine of 25 carcinomas were obtained from premenopausal women, and 16 carcinomas were obtained from postmenopausal women. All cell lines demonstrated positive staining for E1-STS. Interestingly, fine granulated staining of E1-STS on the cell membrane was observed. In addition, Western blotting analysis detected a 65 kD protein with an E1-STS specific band in all breast cancer cell lines regardless of the presence or absence of E2. Twenty-two of 25 (88.0%) carcinomas showed positive staining for E1-STS, whereas negative staining was observed in the interstitial tissue surrounding tumors. In the premenopausal patients, 8 of 10 carcinomas (80.0%) showed positive staining for E1-STS, whereas 14 of 15 carcinomas (93.3%) revealed positive staining in the postmenopausal patients. The frequency of E1-STS expression was relatively higher in postmenopausal patients than in premenopausal patients but not statistically significant. The intensity of immunostaining for E1-STS depended upon the size of the tumor (NS). There was no correlation between E1-STS expression and other parameters. This evidence suggests E1-STS expression may be involved in the development of breast cancer. Further studies are necessary to clarify the relationship between E1-STS expression and prognostic factors. Immunoreactive E1-STS may be localized in cancer cells but not in surrounding tissues in breast cancer.     

组蛋白甲基转移酶SMYD3在乳腺癌中的表达及其意义

目的探讨组蛋白甲基转移酶sMYD3在乳腺癌的表达情况及其意义。方法运用Western blot法检测SMYD3在乳腺癌细胞系的表达,并用免疫组织化学S-P法检测SMYD3在67例乳腺癌组织及10例正常乳腺组织中的表达,最后对sMYD3表达与乳腺癌各临床病理指标行Mann—Whitney U或Kruskal—WallisH检验分析。结果SMYD3在MCF-7、MDA—MB_231、SKBR3和MDA—MB-435s等乳腺癌细胞株中均呈阳性表达,而在正常乳腺组织中均无阳性表达。正常组织与癌性组织之间SMYD3阳性表达的差异有统计学意义（U=130．00,P=0．001）。在乳腺癌组织中,SMYD3的总体阳性率为61．19％。SMYD3阳性表达在年龄,组织病理级别,病理类型,ER、PR、BCL=2、P53表达等亚组间差异均无统计学意义（P值均〉0．050）,而在淋巴结转移、HER2表达亚组间差异有统计学意义（P值均〈0．001）。结论SMYD3阳性表达可作为判断乳腺癌预后的潜在标记物,并有望成为乳腺癌靶向治疗的新靶点。     

A novel dual-target steroid sulfatase inhibitor and antiestrogen: SR 16157, a promising agent for the therapy of breast cancer

Endocrine therapy is the ideal treatment choice for estrogen receptor α (ERα)-positive breast cancer patients. Principal used therapies target either the ERα e.g. by selective ERα modulators (SERMs) such as tamoxifen or target estrogen biosynthesis with aromatase inhibitors. Steroid sulfatase (STS) plays a crucial role in formation of compounds with estrogenic properties, converting inactive sulfate-conjugated steroids to active non-conjugated forms. Steroid sulfates are considered as a reservoir for active steroids due to their prolonged half-life and increased concentration in plasma. STS is present in several tissues including the breast, and the STS the mRNA level and enzyme activity is significantly increased in ERα-positive breast tumors. Inhibition of STS is therefore a new approach for decreasing estrogenic steroids that stimulate breast cancer. The novel dual-acting compound SR 16157 is designed as a sulfamate-containing STS inhibitor that releases a tissue-selective SERM SR 16137. Use of a dual-target STS inhibitor and SERM represents a new strategy in the treatment of hormone-dependent breast cancer. In this study, we tested the potential of SR 16157 and SR 16137 on STS activity, cell growth and ERα function in MCF-7 breast cancer cells. We confirmed that the dual-target compound SR 16157 exerts STS inhibition and antiestrogenic effects. SR 16157 was a highly effective growth inhibitor, being 10 times more potent than the antiestrogens SR 16137 and tamoxifen. Relative to tamoxifen, SR 16137 displays profoundly improved ERα binding affinity and antiestrogenic effects on expression of estrogen-regulated genes. Thus, the dual-target SR 16157 is possibly a promising new treatment alternative, superior to tamoxifen.     

乳腺癌组织P-糖蛋白的表达及其临床意义

目的研究Ｐ糖蛋白（Ｐｇｐ）在乳腺癌组织中的表达及其临床意义。方法用免疫组化法检测６０例乳腺癌、１９例转移性乳腺癌组织Ｐｇｐ表达及化疗疗效。结果乳腺癌Ｐｇｐ表达阳性率为４８．３％,Ｐｇｐ表达与年龄、月经状态、腋淋巴结转移数目、分期、病理组织类型及激素受体状况无关（Ｐ＞０．０５）。Ｐｇｐ阳性组远处转移率（６２．１％）及死亡率（５１．７％）高于阴性组（１６．１％及１２．９％,Ｐ＜０．００５）,５年生存率（４８．３％）低于阴性组（８７．１％,Ｐ＜０．０５）。阳性组辅助化疗后远处转移率（９４．７％）高于阴性组（５７．１％,Ｐ＝０．０４６８）。转移性乳腺癌Ｐｇｐ阳性组化疗有效率（１／１０）低于阴性组（７／９,Ｐ＝０．００５５）。结论免疫组化法检测Ｐｇｐ表达对评估乳腺癌多药耐药、化疗疗效及预后具有一定临床价值。Ｐｇｐ阳性者对ＭＤＲ型药物耐药,疗效及预后差。