目的 探讨原发性肝细胞肝癌（HCC）中O6-甲基鸟嘌呤-DNA甲基转移酶（O6-methylguanine-DNA methyltransferase, MGMT）的表达与临床病理特征以及突变型p53（mutant p53, mtp53）表达的关系。方法 应用免疫组化S-P二步法分别检测10例正常肝组织、40例结节性肝硬化组织和103例人原发性肝细胞性肝癌组织中MGMT和mtp53表达情况。结果 MGMT在正常肝组织、肝硬化和HCC组织中均可表达，正常组表达显著高于肝硬化组（P<0.01）和HCC组（P<0.05），同时HCC组的平均得分又明显高于肝硬化组（P<0.05）。在HCC组织中MGMT表达与mtp53呈显著负相关（P<0.01）。结论 MGMT表达的改变与P53基因突变存在密切联系，对于HCC的发生、发展可能有重要作用，MGMT在肝组织中表达的改变可能作为肝肿瘤易感性标志物。

目的 探讨脱嘌呤/脱嘧啶核酸内切酶（APE1）基因在多发性骨髓瘤（multiple myeloma, MM）细胞中的表达变化及其与马法兰作用的关系。方法 采用免疫细胞化学、Western blot方法检测0~15μmol/L马法兰作用KM3多发性骨髓瘤细胞1~2天后APE1表达的变化，并且通过图像分析系统计算其积分光密度值。结果 马法兰作用可诱导APE1 细胞APE1表达增强，并与其作用时间、剂量成正比。结论 马发兰作用可诱导APE1蛋白表达增强，提示其可能在MM对马发兰耐药中起一定作用。

AIM: To study the effect of caffeic acid phenethyl ester (CAPE) on proliferation, cell cycle, apoptosis and expression of β-catenin in cultured human colorectal cancer (CRC) cell line HCT116. METHODS: HCT116 cells were treated with CAPE at serial concentrations of 80, 40, 20, 10, 5, 2.5 mg/L. The proliferative status of HCT116 cells was measured by using methabenzthiazuron (MTT) assay. Cell cycle was analyzed by using flow cytometry (FCM) with propidium iodide (PI) labeling method. The rate of apoptosis was detected by using FCM with annexin V-FITC and PI double labeling method. β-catenin levels were determined by Western blotting. β-catenin localization in HCT116 was determined by indirect immunofluorescence. RESULTS: After HCT116 cells were exposed to CAPE (80, 40, 20, 10, 5, and 2.5 mg/L) for 24, 48, 72, 96 h, CAPE displayed a strong growth inhibitory effect in a dose- and time-dependent manner against HCT116 cells. FCM analysis showed that the ratio of G0/G1 phase cells increased, S phase ratio decreased and apoptosis rate increased after HCT116 cells were exposed to CAPE (10, 5, and 2.5 mg/L) for 24 h. CAPE treatment was associated with decreased cytoplasmic β-catenin, nuclear β-catenin and a concurrent increase in β-catenin protein expression at cell-cell junctions. CONCLUSION: CAPE could inhibit HCT116 cell proliferation and induce cell cycle arrest and apoptosis. Decreased β-catenin protein expression may mediate the anti-proliferative effects of CAPE.

目的 本研究首先观察N-烷基嘧啶DNA糖基化酶（MPG）在人体骨肉瘤中的表达特点及其与患者预后的关系，进而构建复制缺陷型腺病毒MPG表达载体，探讨人骨肉瘤细胞HOS过表达MPG及其对DNA损伤药物MMS，MNNG和TMZ敏感性的影响。方法 应用免疫组化方法检测60例骨肉瘤组织和3株骨肉瘤细胞中MPG的表达。按染色强度将其分为低表达组和高表达组，统计分析它们与临床病理及患者预后的关系。应用流式细胞术、蛋白印迹和HEX标记寡核苷酸方法确定MPG腺病毒表达载体Ad5 HA-MPG在人体骨肉瘤细胞HOS中的感染效率，MPG蛋白表达和MPG酶活性；MTS、SRB和[3H]胸腺嘧啶掺人法检测细胞存活；PE-Annexinv/7-AAD流式细胞术检测细胞凋亡。结果3株骨肉瘤细胞MPG均呈弱阳性表达。60例骨肉瘤中MPG 40例（65%）为高表达，MPG表达和WHO分型和患者预后显著相关。IO MOI Ad5 HA-MPG可使90%以上HOS感染，感染细胞高表达MPG并具有MPG酶活性。MPG过表达HOS显著地提高DNA损伤性化疗药物MMS、MNNG和TMZ的敏感性，其IC50值分别下降了6.0、4.5和2.5倍。结论Ad5 HA-MPG瞬时MPG过表达，可能是一种提高骨肉瘤细胞对DNA损伤性化疗药物敏感性的潜在治疗方法。

目的 研究DNA损伤修复基因MPG、APEI、XRCC1和MGMT在人体骨肉瘤中的表达特点及其与患者预后的关系，为判估骨肉瘤对化疗的反应性及其预后提供有价值的指标。方法 应用免疫组化方法检测60例骨肉瘤组织和3株骨肉瘤细胞中MPG、APEI、XRCCI和MGMT的表达，按染色强度将其分为低表达组和高表达组。统计分析它们与临床病理及患者预后的关系。结果3株骨肉瘤细胞中APEI和XRCCI呈强阳性，MPG和MGMT呈弱阳性。60例骨肉瘤中MPG、APEI、XRCCI和MGMT高表达分别为40例（65%）、43例（72%）、23例（38%）和32例（53%）。APEI表达和患者年龄，Price's分级和WHO分型有显著性差异（P<0.05），MPG表达和WHO分型有显著性差异（P<0.05）。Spearman秩相关检验显示上述4项指标之间密切相关。单因素相关分析显示APE1和MPG表达与患者预后有显著性差异（P<0.05），COX比例风险模型显示MPG表达与患者预后有关（P<0.01）。结论DNA损伤修复基因MPG、APEI、XRCCI和MGMT的共同表达是骨肉瘤的特点，APEI和MPG表达对于判估骨肉瘤患者的预后可能更有意义。

Osteosarcoma is the most common highly malignant bone tumor with primary appearance during the second and third decade of life. It is associated with a high risk of relapse, possibly resulting from a developed resistance to chemotherapy agents. As a means to overcome osteosarcoma tumor cell resistance and/or to sensitize tumor cells to currently used chemotherapeutic treatments, we examined the role of human apurinic endonuclease 1 (APE1) in osteosarcoma tumor cell resistance and prognosis. Sixty human samples of archived conventional (intramedullary) osteosarcoma were analyzed. APE1 protein was elevated in 72% of these tissues and among those with a known clinical outcome, there was a significant correlation between high APE1 expression levels and reduced survival times. The remaining 28% of samples showed low expression of APE1. Given that APE1 was overexpressed in osteosarcoma, we decreased APE1 levels using silencing RNA (siRNA) targeting technology in the osteosarcoma cell line, human osteogenic sarcoma (HOS), to enhance chemo-and radiation sensitivity. Using siRNA targeted technology of APE1, protein levels were reduced by more than 90% within 24 hours, remained low for 72 hours, and returned to normal levels at 96 hours. There was also a clear loss of APE1 endonuclease activity following APE1-siRNA treatment. A decrease in APE1 levels in siRNA-treated human osteogenic sarcoma cells led to enhanced cell sensitization to the DNA damaging agents: methyl methanesulfonate, H2O2, ionizing radiation, and chemotherapeutic agents. The findings presented here have both prognostic and therapeutic implications for treating osteosarcoma. The APE1-siRNA results demonstrate the feasibility for the therapeutic modulation of APE1 using a variety of molecules and approaches.

目的 探讨DNA损伤修复基因脱嘌呤/脱嘧啶核酸内切酶（apuinic/apyrimidic endonuclease, APE1）在骨肉瘤中的表达及其与肿瘤发生、发展以及血管生成的关系。方法 免疫组化SP法检测三种骨肉瘤细胞株（HOS、U-2OS、Sa-OS-2）、10例正常骨、10例良性骨瘤和60例骨肉瘤组织中APE1蛋白表达，并根据CD34和F VIII-Rag染色结果计数微血管密度（microvessel density, MVD）。结果：1）APE1蛋白在三种骨肉瘤细胞株呈强阳性表达，主要定位于肿瘤细胞的胞核，60例骨肉瘤组织中43例（72%）呈高表达，17例呈低表达，且显著高于正常骨及良性骨瘤，X2=30.997，P=0.000。2）APE1表达的程度和预后密切相关。3）APE1强表达组中的瘤内MVD为46.4±26.7，显著高于弱表达组的33.3±20.7，t=2.277，P=0.030。结论：APE1在骨肉瘤中过度表达与骨肉瘤分化以及MVD密切相关，有可能作为骨肉瘤抗血管生成治疗中一个新的靶向分子。

细胞横固子NF-kB在肿籀血管生成中可能具有核心调节作用，本文选择80例有随访结果的骨肉瘤。应用免疫组化方法检测NF-kB p65亚单位以及血管内皮妇胞生长园子（VEGF）的表达，同时观察肿瘤内微血管密度（MVD），旨在探讨NF-kB表达与骨肉瘤血管生成的关系。结果：80例骨肉瘤中48例NF-kB p65阳性。阳性产物主要定位于肿增细胞浆 NF-kB表运与VEGF表达和MVD呈显著正相差，而且NF-kB阳性组病人存活时闻显著低于阴性组。该结果表明：①NF-kB p65是骨内籀相关的一种癌蛋白，在骨肉瘤的发生和发展中发挥重要的作用；②NF-kB对VEGF可能有正向调节作用，进而影响骨肉瘤血管生成；③NF-kB对肿瘤血管生成的作用，可能是影响骨肉瘤预后的主要原因之一。

目的：探讨血清血管形成相关因子在骨肉瘤组织中的表达，及其与临床病理和微血管密度（MVD）的关系。方法：免疫组化SP法检测80例骨肉瘤组织中VEGF、CD34和F VIII-Rag等蛋白表达，并根据CD34和F VIII-Rag免疫组化染色结果计数MVD。同时采用酶联免疫分析定量检测36例骨肉瘤患者血清中的血管生成因子：血管内皮生成因子（VEGF）、碱性成纤维因子（bFGF）、转化生长因子（TGF-β1）以及内皮抑制素（edostatin, ES）的表达。结果：1）术前血清VEGF（1706ng/dL）和ES（302.4ng/dL）水平明显高于正常对照组，是VEGF的升高与肿瘤的微血高管密度、患者早期复发及其组织高表达密切相关；2）血清VEGF水平（1706ng/dL）和ES（302.4ng/dL）水平呈正相关；3）术后VEGF（490.0ng/dL）和ES（32.7ng/dL）水平明显下降，VEGF复发组较未复发组高，但差异无统计学意义，而复发组的ES水平明显低于未复发组。结论：术前血清VEGF和术后ES水平能反映骨肉瘤血管生成特性，对骨肉瘤早期复发具有重要预测价值，可作为抗血管生成的重要靶标。