乳腺浸润性导管癌中COX-2、MMP-9的表达及临床意义

目的:探讨COX-2和MMP-9蛋白在乳腺浸润性导管癌组织中的表达及其相关性.方法:应用免疫组织化学法检测52例乳腺癌组织COX-2和MMP-9蛋白的表达.结果:乳腺浸润性导管癌组织中COX-2阳性表达率为76.9%(40/52),MMP-9阳性表达率为82.7%(43/52),COX-2阳性表达与患者的年龄、肿瘤大小、雌孕激素受体无明显相关性(P＞0.05),而与淋巴结转移、TNM分期、组织学分级有关(P＜0.05);MMP-9阳性表达与患者的年龄、雌孕激素受体无明显相关性(P＞0.05),而与肿瘤大小、淋巴结转移、TNM分期、组织学分级有关(P＜0.05);乳腺浸润性导管癌组织中COX-2、MMP-9蛋白的表达之间存在显著正相关(r=0.448,P＜0.01).结论:乳腺浸润性导管癌组织中COX-2、MMP-9蛋白高表达,且两者具相关性.COX-2蛋白可通过诱导MMP-9蛋白的表达上调,增加乳腺癌细胞的侵袭力,促进乳腺癌浸润、转移.     

乳腺浸润性导管癌中COX-2、MMP-9的表达及临床意义

目的：探讨COX-2和MMP-9蛋白：在乳腺浸润性导管癌组织中的表达及其相关性。方法-应用免疫组织化学法检测52例乳腺癌组织COX-2和MMP-9蛋白的表达。结果：乳腺浸润性导管癌组织中COX-2阳性表达率为76．9％（40／52）,MMP-9阳性表达率为82．7％（43／52）,COX-2阳性表达与患者的年龄、肿瘤大小、雌孕激素受体无明显相关性（P〉0．05）,而与淋巴结转移、TNM分期、组织学分级有关（P〈0．05）;MMP-9阳性表达与患者的年龄、雌孕激素受体无明显相关性（P〉0．05）,而与肿瘤大小、淋巴结转移、TNM分期、组织学分级有关（P〈0．05）;乳腺浸润性导管癌组织中COX-2、MMP-9蛋白的表达之间存在显著正相关（r=0．448,P〈0．01）。结论：乳腺浸润性：导管癌组织中COX-2、MMP-9蛋白高表达,且两者具相关性。COX-2蛋白可通过诱导MMP-9蛋白如殳达匕调,增加孚嘣塘细胞的侵袭力,促进乳腺癌浸润、转移。     

乳腺浸润性导管癌组织中环氧合酶-2的表达与细胞增生和血管生成的关系

目的 探讨环氧合酶-2(COX-2)蛋白在乳腺浸润性导管癌组织中的表达及其与细胞增生和微血管密度(MVD)的关系.方法 应用免疫组织化学法检测52例乳腺浸润性导管癌组织中COX-2和Ki-67蛋白的表达情况,用CD34抗体标记乳腺癌血管内皮细胞,计算MVD.结果 乳腺浸润性导管癌组织中COX-2蛋白的阳性表达率为57.7%,COX-2阳性组Ki-67指数为(56.07±17.37)%,COX-2阴性组Ki-67指数为(27.32±16.28)%,COX-2阳性组MVD为32.46±12.59,COX-2阴性组MVD为25.04±10.48,COX-2阳性组Ki-67、MVD显著高于COX-2阴性组(P＜0.05).结论 乳腺浸润性导管癌组织中COX-2蛋白高表达,COX-2可促进肿瘤细胞增生和肿瘤肿瘤血管生成,从而成为促进乳腺癌浸润、转移的途径之一.     

Maspin和MMP-2在乳腺浸润性导管癌中的表达及其临床意义

目的探讨Maspin和MMP-2在乳腺浸润性导管癌组织中的表达及意义。方法应用免疫组织化学SP法检测60例乳腺浸润性导管癌及20例正常乳腺组织中Maspin和MMP-2的表达。结果Maspin在乳腺浸润性导管癌中表达阳性率为45.0%,低于在正常乳腺组织中的表达阳性率95.0%(P<0.05);MMP-2在乳腺浸润性导管癌中表达阳性率为75.0%,高于在正常乳腺组织中的表达阳性率15.0%(P<0.05)。Maspin和MMP-2的表达均与乳腺浸润性导管癌的淋巴结转移相关(P<0.05),MMP-2的表达还与乳腺浸润性导管癌临床分期相关(P<0.05)。乳腺浸润性导管癌中Maspin与MMP-2的表达呈负相关(r=-0.11,P<0.05)。结论 Maspin对乳腺浸润性导管癌的转移有抑制作用,MMP-2对乳腺浸润性导管癌的转移有促进作用,检测Maspin和MMP-2的表达可以成为判断乳腺浸润性导管癌浸润及转移能力的重要指标之一。     

乳腺癌组织中CD44v6和MMP-2的表达及其与淋巴结转移的关系

目的:检测CD44v6、MMP-2在乳腺浸润性导管癌中的表达情况,探讨它们与淋巴结转移的关系。方法:采用免疫组化SP法检测100例乳腺浸润性导管癌中CD44v6、MMP-2的表达情况。结果:CD44v6蛋白在正常乳腺组织及乳腺浸润性导管癌中的阳性表达率分别为12.5%(5/40)、87%(87/100),CD44v6在淋巴结转移组中的阳性表达率(56/60)明显高于无淋巴结转移组(P<0.05)。MMP-2蛋白在正常乳腺组织及乳腺浸润性导管癌中的阳性表达率分别为0(0/40)、94%(94/100),MMP-2在淋巴结转移组中的阳性表达率(59/60)明显高于无淋巴结转移组(P<0.05)。结论:CD44v6、MMP-2蛋白在乳腺浸润性导管癌组织高表达,且均与淋巴结转移有关(P<0.05),联合检测CD44v6与MMP-2有助于综合判断乳腺浸润性导管癌的恶性程度和转移潜能。     

uPA和HIF-1α在乳腺浸润性导管癌中的表达及其相关性

目的:探讨uPA和HIF-1α蛋白在乳腺浸润性导管癌组织中的表达及其相关性.方法:应用免疫组织化学法检测52例乳腺浸润性导管癌组织中uPA、HIF-1α蛋白表达.结果:乳腺浸润性导管癌组织中uPA蛋白阳性表达率为57.7%,HIF-1α蛋白阳性表达率为82.7%,uPA、HIF-1α蛋白表达之间存在显著正相关,r=0.498,P<0.01.结论:乳腺浸润性导管癌组织中存在uPA、HIF-1α的高表达,且两者之间的表达强度具有相关性.uPA蛋白可通过诱导HIF-1α蛋白的表达上调,成为促进乳腺癌侵袭、转移的途径之一.     

COX-2在乳腺癌组织的表达及其与临床病理参数的相关性

目的:评价乳腺浸润性导管癌组织COX-2蛋白的表达情况及其与相关临床或病理参数间的相关性.方法:取101例资料完整的乳腺浸润性导管癌手术切除标本为研究对象,免疫组化方法检测COX-2、EGFR、cerbB-2等相关蛋白表达,采用χ2检验评估COX-2表达与不同的临床或病理参数间的关系.结果:肿瘤细胞中COX-2蛋白免疫组化阴性者15例(14.9%);阳性者86例(85.1%),COX-2高表达与乳腺癌脉管侵犯、淋巴结侵犯、分期、PR表达、cerbB-2表达及p53表达具有相关性.结论:乳腺浸润性导管癌中COX-2表达水平与肿瘤转移及分期均密切相关.     

乳腺癌组织中CD44v6和MMP-2的表达及其与淋巴结转移的关系

目的：检测CD44v6、MMP-2在乳腺浸润性导管癌中的表达情况,探讨它们与淋巴结转移的关系。方法：采用免疫组化SP法检测100例乳腺浸润性导管癌中CD44v6、MMP-2的表达情况。结果：CD44v6蛋白在正常乳腺组织及乳腺浸润性导管癌中的阳性表达率分别为12．5％（5／40）、87％（87／100）,CD44v6在淋巴结转移组中的阳性表达率（56／60）明显高于无淋巴结转移组（P〈0．05）。MMP-2蛋白在正常乳腺组织及乳腺浸润性导管癌中的阳性表达率分别为0（0／40）、94％（94／100）,MMP-2在淋巴结转移组中的阳性表达率（59／60）明显高于无淋巴结转移组（P〈0．05）。结论：CD44v6、MMP-2蛋白在乳腺浸润性导管癌组织高表达,且均与淋巴结转移有关（P〈0．05）,联合检测CD44v6与MMP-2有助于综合判断乳腺浸润性导管癌的恶性程度和转移潜能。     

uPA和HIF-1α在乳腺浸润性导管癌中的表达及其相关性

目的：探讨uPA和HIF-1α蛋白在乳腺浸润性导管癌组织中的表达及其相关性。方法：应用免疫组织化学法检测52例乳腺浸润性导管癌组织中uPA、HIF-1α蛋白表达。结果：乳腺浸润性导管癌组织中uPA蛋白阳性表达率为57.7%,HIF-1α蛋白阳性表达率为82.7%,uPA、HIF-1α蛋白表达之间存在显著正相关,r=0.498,P〈0.01。结论：乳腺浸润性导管癌组织中存在uPA、HIF-1α的高表达,且两者之间的表达强度具有相关性。uPA蛋白可通过诱导HIF-1α蛋白的表达上调,成为促进乳腺癌侵袭、转移的途径之一。     

COX-2在乳腺癌组织的表达及其与临床病理参数的相关性

目的：评价乳腺浸润性导管癌组织COX-2蛋白的表达情况及其与相关临床或病理参数间的相关性。方法：取101例资料完整的乳腺浸润性导管癌手术切除标本为研究对象,免疫组化方法检测COX-2、EGFR、cerbB-2等相关蛋白表达,采用X。检验评估COX-2表达与不同的临床或病理参数间的关系。结果：肿瘤细胞中COX-2蛋白免疫组化阴性者15例（14．9％）;阳性者86例（85．1％）,COX-2高表达与乳腺癌脉管侵犯、淋巴结侵犯、分期、PR表达、cerbB-2表达及p53表达具有相关性。结论：乳腺浸润性导管癌中COX-2表达水平与肿瘤转移及分期均密切相关。     

Antitumor activity of 2'-deoxy-2'-methylidenecytidine, a new 2'-deoxycytidine derivative

The antitumor activity of 2'-deoxy-2'-methylidenecytidine (DMDC), an inhibitor of DNA synthesis, was examined and compared with that of 1-beta-D-arabinofuranosylcytosine (ara-C) against various murine tumors and human tumor xenografts. Against P388 murine leukemia, repeated treatments of DMDC were more effective than its single administration. Interestingly, DMDC was effective against colon 26 murine carcinoma, M5076 murine reticulum cell sarcoma, LX-1 human lung cancer xenograft, and SK-Mel-28 human melanoma xenograft, which are less sensitive or refractory to ara-C, while DMDC was not more potent against murine leukemias P388 and L1210 than ara-C. The in vitro cytotoxic effects of DMDC and ara-C against L1210 leukemia cells were prevented dose dependently by deoxycytidine, suggesting that DMDC, like ara-C, may require phosphorylation by deoxycytidine kinase for antitumor activity. DMDC was effective against human and murine experimental tumor models, especially nonleukemic tumors refractory to ara-C, suggesting that DMDC will be a promising agent for the treatment of cancer.     

转染和干扰Runx2基因对K7M2细胞的影响

 目的探讨干扰和转染Runx2基因后对K7M2骨肉瘤细胞体外生物学活性的影响。方法分别将阴性对照质粒、携带Runx2和siRNA Runx2的质粒转染到K7M2细胞中。应用Western blot检测转染和干扰Runx2基因对其蛋白表达的影响,并检测细胞体外的增殖能力(MTT )、侵袭性、迁移性和黏附性的变化,并应用流式细胞仪（FCM）检测相应细胞的凋亡率和细胞周期分布的变化。结果与空白对照组比,转染Runx2和siRunx2组的K7M2细胞中Runx2蛋白表达分别明显增强和减弱,而阴性对照质粒组没有明显改变。转染siRunx2组的细胞,MTT结果显示增殖能力明显下降,迁移性、侵袭性和增殖指数（PI）也明显下降,与空白对照组比差异有统计学意义（P<0.05）,黏附性和凋亡率却没有明显变化。转染Runx2组K7M2的细胞增殖能力、迁移性、侵袭性和PI明显增高,与空白对照组比差异有统计学意义（P<0.05）,黏附性和凋亡率没有明显变化。阴性对照组和空白对照组比上述指标没有明显改变。结论Runx2可以促进K7M2细胞增殖,增强细胞的侵袭、迁移能力,其机制可能是和改变细胞周期的分布,使细胞更快的进入G₂/M期有关。     

Identification of CYP2C9*2 Allele in HepG2 Cell Line

Background

Hepatoma is caused by many factors including alcohol, chemicals, viral infection, and chronic inflammation. Cytochrome P450 polymorphism plays an important role in its pathogenesis. CYP2D6, CYP2E1, and CYP1A1 have been identified to be related with hepatic carcinogenesis and tumor size and stage. However, no studies have been performed on CYP2C9, a major CYP in the liver and hepatoma.

Aim of the study

To identify if there is polymorphism of CYP2C9 in a HepG2 cell line.

Methods

A pair of primers was used to clone CYP2C9 exon 3 region and subsequently sequenced. The sequence was compared to normal CYP2C9 for identification of any mutation.

Results

A point mutation was identified. It was located in the amino acid number 144 of CYP2C9 protein with the change of normal amino acid arginine into cysteine, which is the same as identified in poor metabolism patients as homozygous CYP2C9*2.

Conclusions

There is a mutation (CYP2C9*2/ CYP2C9*2) in a HepG2 cell line. Thus, polymorphism of CYP2C9 may also be involved in the carcinogenesis of hepatoma as CYPs2D6 and 2E1.     

Cyclooxygenase-2 (COX-2)-independent anticarcinogenic effects of selective COX-2 inhibitors

Nonsteroidal antiinflammatory drugs (NSAIDs) appear to reduce the risk of developing cancer. One mechanism through which NSAIDs act to reduce carcinogenesis is to inhibit the activity of cyclooxygenase-2 (COX-2), an enzyme that is overexpressed in various cancer tissues. Overexpression of COX-2 increases cell proliferation and inhibits apoptosis. However, selective COX-2 inhibitors can also act through COX-independent mechanisms. In this review, we describe the COX-2-independent molecular targets of these COX-2 inhibitors and discuss how these targets may be involved in the anticarcinogenic activities of these selective COX-2 inhibitors. We also compare the concentrations of these inhibitors used in in vitro and in vivo experiments and discuss the implications of the in vitro studies for clinical management of cancer with these drugs.     

Evaluation of the antitumor activity of gemcitabine (2',2'-difluoro-2'-deoxycytidine)

A new pyrimidine antimetabolite, 2',2'-difluorodeoxycytidine, Gemcitabine (LY188011, dFdCyd) has been synthesized and evaluated in experimental tumor models. dFdCyd is a very potent and specific deoxycytidine analogue. The concentration required for 50% inhibition of growth is 1 ng/ml in the CCRF-CEM human leukemia cell culture assay. Concurrent addition of deoxycytidine to the cell culture system provides about a 1000-fold decrease in biological activity. The inhibition of growth of human leukemia cells in culture led to the in vivo evaluation of this compound as a potential oncolytic agent. Maximal activity in vivo was seen with dFdCyd when administered on an every third day schedule. 1-beta-D-Arabinofuranosylcytosine, administered on a daily for 10-day schedule, was directly compared to dFdCyd in this evaluation. dFdCyd demonstrated good to excellent antitumor activity in eight of the eight murine tumor models evaluated. 1-beta-D-Arabinofuranosylcytosine was substantially less active or had no activity in these same tumor models. This in vivo activity against murine solid tumors supports the conclusion that dFdCyd is an excellent candidate for clinical trials in the treatment of cancer.     

Cell cycle-dependent phosphorylation of Disabled-2 by cdc2

Disabled-2 (Dab2; also known as p96 and DOC-2) is a signal transduction protein that has been implicated in the control of cell growth. Dab2 is known to be a phosphoprotein, but little is known about the kinases that phosphorylate Dab2. We have found that Dab2 phosphorylation is markedly increased during the mitosis phase of the cell cycle. This phosphorylation is blocked by roscovitine, a selective inhibitor of cyclin-dependent kinases. Dab2 robustly coimmunoprecipitates from cells with the cyclin-dependent kinase cdc2, and purified cdc2 can phosphorylate purified Dab2 fusion proteins in vitro on multiple sites. Cellular phosphorylation of Dab2 by cdc2 promotes the association of Dab2 with Pin1, a peptidylprolyl isomerase that regulates the rate of Dab2 dephosphorylation. These findings reveal that Dab2 is differentially phosphorylated during the cell cycle by cdc2 and provide a potential feedback mechanism by which Dab2 inhibition of cell growth and proliferation may be regulated.     

Polymorphisms in prostaglandin synthase 2/cyclooxygenase 2 (PTGS2/COX2) and risk of colorectal cancer

Inflammation plays a key role in the development of colorectal cancers. We have investigated the relationship between PTGS2 (COX2) polymorphisms and colorectal cancer risk in a hospital based case-control study. We recruited 292 patients with colorectal cancer and 274 controls from new patients admitted to Bellvitge Hospital, Barcelona, Spain, from 1996 to 1998. Subjects responded to a questionnaire on risk factors. Genotypes of the eight more frequent polymorphisms of PTGS2 were determined. Two polymorphisms are located in the promoter sequence, one in the untranslated region of exon 1, one in exon 3, one in intron 5, two in the untranslated region of exon 10, and one downstream of the last polyadenylation (poly-A) signal. Associations were analysed with logistic regression models assuming a dominant effect for rare variants to increase statistical power. An association was detected between colorectal cancer and a polymorphism in the untranslated region of exon 10 of PTGS2, with an odds ratio (OR) of 2.49, 95% confidence interval (CI) of 1.17-5.32, P=0.01. A nearby polymorphism downstream of the last poly-A signal also showed a nonsignificant increase in risk (OR 2.17, 95% CI 0.99-4.78, P=0.05). Analysis of haplotypes confirmed that individuals with these variants were at increased risk of colorectal cancer (OR compared to the most frequent haplotype: 2.17, 95% CI 0.97-4.84, P=0.06) Interactions between PTGS2 genotype and use of nonsteroidal anti-inflammatory drugs and risk of colorectal cancer were also explored.