晚期肾癌靶向治疗的作用机制 耐药机制与对策

随着对肾癌发生、发展过程中分子调控机制研究的不断深入,以分子信号通路中的关键蛋白酶/ 蛋白作为靶点开发靶向药物,较好地改善了晚期肾癌患者的治疗现状。但是靶向药物在治疗过程中常出现耐药,从而导致治疗效果降低。本文将对肾癌靶向治疗的作用机制、耐药机制与对策作简要综述。     

Anti-tumor Effects of Interleukin-4 and Interleukin-5 against Mouse B Cell Lymphoma and Possible Mechanisms of Their Action

We investigated the anti-tumor effects of recombinant mouse interleukin (IL)-4 and IL-5 by using a transplantable B cell lymphoma 38C13 cell line as a model. Daily local administration of either IL-4 or IL-5 produced moderate but significant inhibition of the rate of local tumor growth and prolongation of mean survival time (MST) in syngeneic C3H/HeJ mice; these anti-tumor effects appeared to plateau at low doses. Histopathologic and immuno-histochemical examination revealed necrotic changes in the cytokine-treated tumors, associated with infiltration of inflammatory cells such as eosinophils, macrophages, and lymphocytes. The infiltrating lymphocytes were found to be Thy-1.2⁺ T cells. To elucidate the importance of T cells, the rate of tumor growth and the MSTs were compared between athymic T cell-deficient BALB/c nude mice and immunocompetent C3H/HeJ mice. In the nude mice the transplanted tumor grew more rapidly and the MST was shorter than in the normal mice, suggesting a significant contribution of infiltrating T cells in the anti-tumor effects of the interleukins. Lastly, in vitro, growth inhibition of the 38C13 cells was observed in a dose-dependent manner at relatively high concentrations of either cytokine. Therefore, we conclude that both IL-4 and IL-5 have moderate anti-tumor effects against 38C13 B cell lymphoma both in vivo and in vitro , and that the observed in vivo anti-tumor effects are probably mediated both by tumoristatic action of infiltrating cells, such as eosinophils, macrophages and T lymphocytes, and by direct anti-proliferative action of the recombinant cytokines.     

How Cancers Escape Immune Destruction and Mechanisms of Action for the New Significantly Active Immune Therapies: Helping Nonimmunologists Decipher Recent Advances

With the Food and Drug Administration and other worldwide regulatory authorities’ approval of ipilimumab (Yervoy), sipuleucel-T (Provenge), nivolumab (Opdivo), and pembrolizumab (Keytruda), oncologic therapy has now moved into noncancer cell targets within the immune system. For many nonimmunologists, understanding how these vastly different therapies work to improve survival, like no other therapies have in the past, is a challenge. The present report reviews the normal function of the immune system, how cancers escape the normal immune system, and how these new therapies improve immune system reactions against cancers.

Implications for Practice:

Oncologists have tremendous experience with therapies that target the cancer cells. New biologic agents have been rapidly introduced recently that target not cancer cells, but the patient’s immune cells. The mechanisms of action of these immune-based biologic agents are within the host immune system. To understand these new biologic therapies, basic knowledge of normal and abnormal immune function is essential. The present report explains the up-to-date basic immune normal and abnormal function and prepares the oncologist to understand how the new drugs work, why they work, and why there are associated adverse events.     

Prognostic significance of matrix metalloproteinases-1, -2, -7 and -13 and tissue inhibitors of metalloproteinases-1, -2, -3 and -4 in colorectal cancer

Strong expression of many matrix metalloproteinases (MMPs) has been related to poor survival of colorectal cancer (CRC) patients. The expression of tissue inhibitors of metalloproteinases (TIMPs) has been associated with both a beneficial and a poor outcome and there is thus a need to further clarify the significance of MMPs and TIMPs in CRC. The prognostic significance of 4 MMPs and TIMPs in CRC was evaluated. Formalin-fixed, paraffin-embedded tissue arrayed samples of 351 patients with primary colon or rectal cancer of Dukes' stages A-D were selected for immunohistochemical staining of MMP-1, -2, -7 and -13, and TIMP-1, -2, -3 and -4. High expression of MMP-2 in the malignant epithelium as well as in the surrounding stroma was associated with reduced survival of colon cancer patients. Strong epithelial and stromal cytoplasmic staining of TIMP-3 was associated with a longer survival in rectal cancer patients, and here the interobserver variation for evaluating the degree of staining was lower than for epithelial staining. Strong stromal cytoplasmic staining of TIMP-4 predicted longer survival of rectal cancer patients. Multivariate analysis showed that stromal cytoplasmic TIMP-3 staining was the only marker of independent prognostic value. MMP-2 might be a useful prognostic marker in colon cancer, and TIMP-3 and TIMP-4 in rectal cancer, but the findings associated with stromal staining should be interpreted with some caution. Different biologic behavior or different genetic development may explain the differences between colon and rectal cancers regarding the expression of MMP-2, TIMP-3 and TIMP-4.     

银莲花素A对S180、H22和U14细胞小鼠移植瘤的抑制作用

背景与目的:有研究证明,从两头尖中分离得到的三萜皂甙银莲花素A在体外具有较好的抑瘤活性.本研究将观察银莲花素A在体内对小鼠移植瘤的抑制活性.方法:MTT法检测银莲花素A对人鼻咽癌KB细胞和人卵巢癌SKOV3细胞的抑制率.体内实验选取小鼠肉瘤S180细胞、小鼠肝癌H22细胞和小鼠宫颈癌U14细胞.观察银莲花素A注射给药对S180、H22和U14细胞小鼠移植瘤的抑瘤率,以及灌胃给药对小鼠肉瘤S180移植瘤的抑瘤率.测定银莲花素A的急性毒性.结果:银莲花素A在体外对KB细胞和SKOV3细胞的IC50分别为4.64 μg/mL和1.40 μg/mL.4.5 mg/kg银莲花素A腹腔注射对S180、H22和U14细胞小鼠移植瘤的抑瘤率分别为60.5%、36.2%和61.8%.200 mg/kg灌胃给药时抑瘤率为64.7%.灌胃给药和腹腔注射银莲花素A的LD50分别为1.1 g/kg和16.1 mg/kg.结论:银莲花素A在体内外均具有较好的抑瘤作用,体内可抑制小鼠移植瘤的生长,是一个有潜力的抗癌药物.     

Involvement of HLA Antigens, Interleukin 2 Receptor and B-Cell Growth Factor in Interferon Action in Hairy Cell Leukemia

Little is known about the mechanism(s) by which alpha-interferon (aIFN), when used as a biotherapeutic agent, suppresses the malignant cells and restores the normal phenotype of cells in patients with hairy cell leukemia (HCL). In previous studies using scanning electron microscopy (SEM) we found that alFN induced unique membrane alterations in target hairy cells in vitro. In addition, aIFN was shown to enhance the expression of HLA class II antigens on HCL cells, to induce the production of new proteins in such cells, and to lower the high levels of soluble IL-2 receptors in the serum of HCL patients. In the light of these results, and the fact that restoration of natural killer cell activity occurs in aIFN-treated patients well after the hematologic profile begins to improve, our studies have focused on the hypothesis that IFNs act directly on the target malignant cells, leading to the elimination of these cells either by inhibiting the proliferation of the malignant cells and/or triggering changes in the differentiation status of the malignant cells that lead to suppression (cytoconversion) of the malignant phenotype. We review the current hypotheses regarding alFN action on leukemic cells, with special reference to its potential antagonism with BCGF     

Mantle cell lymphomas acquire increased expression of CCL4, CCL5 and 4-1BB-L implicated in cell survival

We have analyzed mantle cell lymphomas (MCLs), using high-density DNA microarrays, and confirmed the expression of differentially regulated antigens, using flow cytometry and immunohistochemistry. The results show that MCLs acquire expression of molecules that normally are involved in interaction with other immune cells and, thus, might affect the ability of the tumor to survive. The MCL signature is represented by the overexpression of the chemokine CCL4 (MIP-1beta), implicated in the recruitment of regulatory T cells, as well as CCL5 and 4-1BB-L. The latter molecules are normally involved in chemotaxis of T cells and B cell activation, respectively. Signaling through 4-1BB-L allows B cells to proliferate and the expression of its ligand, by the intra-tumoral mesh of follicular dendritic cells (FDC), could thus serve as a paracrine loop facilitating growth and survival of MCL cells.     

细胞周期相关基因CDKN2A、TP53、RB1 和BRCA2在恶性肿瘤中的研究进展

CDKN2A、TP53、RB1和BRCA2是细胞周期相关基因,主要通过转录合成细胞周期相关蛋白,在细胞增殖和凋亡的调控方面发挥重要作用。并且CDKN2A、TP53、RB1和BRCA2基因在多种肿瘤的发生发展以及肿瘤的治疗和预后方面扮演重要角色。本文就近年来对CDKN2A、TP53、RB1和BRCA2在肿瘤中作用以及对患者治疗疗效和预后影响的研究进展进行综述。     

哈萨克族食管癌中MMP-1、MMP-2、MMP-7、TIMP-1和MTA1的表达及其临床病理意义

目的：探讨在新疆哈萨克族食管癌组织中基质金属蛋白酶1、2、7（MMP-1、MMP-2、MMP-7）、基质金属蛋白酶抑制因子1（TIMP-1）和肿瘤转移相关基因1（MTA1）mRNA的表达及其临床意义。方法：采用RT-PCR方法检测75例哈萨克族食管癌标本中MMP-1、MMP-2、MMP-7、TIMP-1和MTA1 mRNA的表达水平,分析其表达与临床病理特征的关系。结果：MMP-1、MMP-2、MMP-7、TIMP-1和MTA1 mRNA在哈萨克族食管癌组织中的表达较正常组织增高,差异均具有统计学意义（P均〈0.05）;且MMP-2、MMP-7、MTA1 mRNA的表达与淋巴结转移有关（P〈0.05）,MMP-1、MMP-7 mRNA的表达与临床分期有关（P〈0.05）;TIMP-1与MMP-1、MMP-7、MTA1的表达呈正相关（r=0.446、0.458、0.333,P均〈0.01）。结论：MMP-1、MMP-2、MMP-7、TIMP-1和MTA1 mRNA表达上调共同参与了哈萨克族食管癌的发生发展过程;MMP-2、MMP-7和MTA1可能是哈萨克族食管癌发生侵袭、转移的主导因素。     

CDK1、CDK2 siRNA干扰对肿瘤细胞凋亡和细胞周期的影响

Objective: We investigated the influence of CDK1 and CDK2 expression inhibited by cotransfection of CDK1 and CDK2 siRNA on cell cycle and apoptosis, explored the exact role of cell cycle master regulator in tumor cell apoptosis process. Methods: The siRNA targeting the CDK1 and CDK2 genes were synthesized and simultaneously cotransfected into Hela cells by lipofectamine 2000.48 or 60 h after the cotransfection, CDK1 and CDK2 protein expressions were examined by Western blot. Cell cycle distribution was analyzed by flow cytometry. Cell apoptosis was detected by the Annexin V/PI method. The changes of the transfected cell morphological under a microscope after Wright-Giemsa Staining were studied. Results: CDK1 and CDK2 protein expression was decreased at 48 or 60 h after cotransfection. The accumulation of the G2/M and S phase population in cell cycle of the cotrensfected cells at 48 or 60 h after transfection was enhanced obviously compared with control. The ratio of apoptotic cell of cotransfected cells at 48 or 60 h after transfection was increased significantly compared with control. More binucleate or multinucleate cells among cotransfected cells were observed under the microscope. Conclu- sion: The decreased expression of CDK1 and CDK2 by cotransfection of CDK1 and CDK2 siRNA not only leads to tumor cell cycle arrest in S phase and G2/M phase, but also induces tumor cell apoptosis.     

COX-2、c-erbB-2、nm23-H1、PCNA表达与宫颈鳞癌预后的关系

 目的　探讨环氧合酶 2 (cyclooxygenase 2 ,COX 2 )、c erbB 2、nm2 3 H 1、增殖细胞核抗原 (prolifer atingcellnuclearantigen ,PCNA)的表达与宫颈鳞癌预后的关系。 方法　应用免疫组化S P方法检测了3 0例正常宫颈和 52例宫颈鳞癌组织中COX 2、c erbB 2、nm 2 3 H1、PCNA的表达。结果　 (1)COX 2、c erbB 2表达与淋巴结转移无关 (P >0 .0 5) ,而PCNA表达与淋巴结转移呈正相关 (P <0 .0 5) ,nm 2 3 H1表达与淋巴结转移呈负相关 (P <0 .0 5) ;(2 )复发患者COX 2与PCNA表达阳性率明显高于未复发的患者 (P <0 .0 5) ,而nm2 3 H 1及c erbB 2表达与复发无关 (P >0 .0 5) ;(3 )COX 2、PCNA表达与预后呈负相关 ,而nm2 3 H1表达则与预后呈正相关 ,c erbB 2表达与生存无关。多因素分析表明淋巴结转移、COX 2、nm 2 3 H1是影响宫颈鳞癌预后的独立因素。结论　COX 2、nm...     

MT1-MMP和TIMP-2在非小细胞肺癌中的表达及与病理特征的相关性

目的 研究NSCLC中MT1-MMP及TIMP-2的表达及与病理类型分型、淋巴结转移和TNM分期关系.方法 采用免疫组化SP法检测MT1-MMP及TIMP-2在80例NSCLC患者和80例癌旁组织中的表达.结果 在NSCLC中MT1-MMP阳性表达率为73.8%,癌旁组织中为52.5%,(P<0.05).在NSCLC中TIMP-2的阳性表达率为42.5%,癌旁组织为72.5%,(P<0.05).MT1-MMP和TIMP-2的表达与肺癌的病理分类及肿瘤大小无关,而与TNM分期及淋巴结转移有关.MT1-MMP和TIMP-2的表达呈负相关(r=-0.635,P<0.05).结论 MT1-MMP和TIMP-2可能参与了NSCLC的发生发展过程,有望成为判断肺癌转移和预后的指标.     

MT1-MMP和TIMP-2在非小细胞肺癌中的表达及与病理特征的相关性

目的：研究NSCLC中MT1-MMP及TIMP-2的表达及与病理类型分型、淋巴结转移和TNM分期关系。方法：采用免疫组化SP法检测MT1-MMP及TIMP-2在80例NSCLC患者和80例癌旁组织中的表达。结果：在NSCLC中MT1-MMP阳性表达率为73.8%,癌旁组织中为52.5%,（P〈0.05）。在NSCLC中TIMP-2的阳性表达率为42.5%,癌旁组织为72.5%,（P〈0.05）。MT1-MMP和TIMP-2的表达与肺癌的病理分类及肿瘤大小无关,而与TNM分期及淋巴结转移有关。MT1-MMP和TIMP-2的表达呈负相关（r=-0.635,P〈0.05）。结论：MT1-MMP和TIMP-2可能参与了NSCLC的发生发展过程,有望成为判断肺癌转移和预后的指标。     

人脑胶质瘤中Smad-1、Smad-2和Smad-4的表达及意义

目的 检测BMPs/BMPR/Smads信号转导通路上的人脑胶质瘤组织中Smad-1、Smad-2、Smad-4的表达,探讨其在人脑胶质瘤发生发展中的作用及其与临床病理分级的关系。方法 分别应用RT-PCR技术和免疫组织化学SABC法检测20例正常脑组织和40例不同级别人脑胶质瘤组织中Smad-1、Smad-2、Smad-4 mRNA和蛋白质表达量,并分析其与患者的年龄、性别、病理分级的相关性。结果 人脑胶质瘤组织Smad-1、Smad-2、Smad-4 mRNA表达量比正常脑组织显著降低（0.277±0.125 vs.0.573±0.097,P<0.01;0.282±0.111 vs. 0.613±0.105,P<0.01;0.389±0.101 vs. 0.483±0.098,P<0.05）,且Ⅲ+Ⅳ组显著低于Ⅰ+Ⅱ组（0.250±0.106 vs. 0.327±0.119,P<0.05;0.2451±0.109 vs. 0.315±0.113,P<0.05;0.347±0.121 vs. 0.434±0.102,P<0.05）。蛋白质表达的阳性率显著低于正常脑组织（37.50%vs. 75.00%,P<0.0 1;20.00% vs. 65.00%,P<0.01; 25.00% vs. 70.00%,P<0.01）,且Ⅲ+Ⅳ组显著低于Ⅰ+Ⅱ组（16.67% vs. 54.55%,P<0.05;5.56% vs. 31.82%, P<0.05;5.56% vs. 40.91%,P<0.05）。上述指标的变化与患者的年龄及性别无关。结论 Smad-1、Smad-2、Smad-4 在人脑胶质瘤中的表达水平与胶质瘤的发生发展和恶性程度密切相关,提示Smad-1、Smad-2、Smad-4可能参与胶质瘤发生发展过程。     

基质金属蛋白酶及其抑制剂在乳腺癌中的表达及其临床意义

背景与目的:基质金属蛋白酶(matrixmetalloproteinase,MMP)与基质金属蛋白酶组织抑制剂(tissueinhibitorofmatrixmetalloproteinase,TIMP)的表达失平衡在肿瘤侵袭、转移过程中起重要作用,但与乳腺癌预后关系的报道少见。本研究探讨MMP-2、MMP-9和TIMP-1、TIMP-2的表达与乳腺癌侵袭、转移和预后的关系。方法:原位杂交、免疫组化检测66例有临床和随访资料的乳腺癌患者的MMP-2mRNA、TIMP-2mRNA和MMP-2、MMP-9、TIMP-1、TIMP-2蛋白表达。统计学分析采用χ2检验、Kaplan-Meier和Cox多因素回归分析。结果MMP-2mRNA、TIMP-2mRNA和MMP-2、MMP-9、TIMP-1TIMP-2蛋白的阳性表达率分别为66.7%(44/66)、65.2%(43/66)和71.2%(47/66)、68.2%(45/66)、40.9%(2766)、69.7%(46/66),其中MMP-2蛋白与MMP-2mRNAMMP-9蛋白及TIMP-2mRNA与TIMP-2蛋白的表达存在显著性正相关(P<0.01);TIMP-1与MMP-9蛋白表达呈负相关(P<0.01)。有淋巴结转移的乳腺癌中MMP-2、MMP-9蛋白表达显著高于无转移者,但TIMP-2mRNA、TIMP-1蛋白表达显著低于无转移者(P<0.05)。乳腺癌中MMP-2mRNA和MMP-9蛋白表达与肿块大小、生存状况有显著性相关(P0.05),此外,MMP-9蛋白表达与临床分期存在正相关性(P<0.01)。绝经和ER表达阴性患者的MMP-2mRNA表达水平增高(P<0.     

ANGPTL4在调控骨巨细胞瘤细胞增殖、血管生成和破骨分化作用的实验研究

目的探讨人血管生成素样蛋白4(ANGPTL4)基因沉默对骨巨细胞瘤单核基质细胞(GCTSC)增殖、血管生成以及破骨分化的作用。方法体外培养GCTSC细胞系,采用ANGPTL4 shRNA、Scrambled shRNA转染作为ANGPTL4 shRNA组和阴性对照组,另设置无处理的空白对照组。采用实时荧光定量PCR(QPCR)和Western blotting法检测ANGPTL4 mRNA和蛋白表达。MTT法和流式细胞术检测细胞增殖和凋亡。将GCTSC细胞分别与人脐静脉内皮细胞(HUVECs)、小鼠骨髓单核细胞(BMM)共培养,观察HUVECs成管能力和BMM破骨分化能力。结果与空白对照组和阴性对照组比较,ANGPTL4 shRNA组ANGPTL4 mRNA(0.174±0.045)和蛋白表达量(0.098±0.020)均明显降低;而ANGPTL4 shRNA组GCTSC细胞增殖活性降低,凋亡率升高(P<0.05)。与HUVECs共培养后,ANGPTL4 shRNA组闭合管数量[(57.35±17.24)%]减少(P<0.05);与BMM共培养后,ANGPTL4 shRNA组TRAP染色阳性的破骨细胞样多核巨细胞数量[(48.36±21.79)%]减少(P<0.05)。结论沉默骨巨细胞瘤GCTSC细胞ANGPTL4基因表达后,细胞增殖活性降低,凋亡率增加,并且对肿瘤血管生成和多核巨细胞破骨分化有一定的抑制作用。     

Potentially functional genetic variants in PLIN2, SULT2A1 and UGT1A9 genes of the ketone pathway and survival of nonsmall cell lung cancer

The ketone metabolism pathway is a principle procedure in physiological homeostasis and induces cancer cells to switch between glycolysis and oxidative phosphorylation for energy production. We conducted a two-phase analysis for associations between genetic variants in the ketone metabolism pathway genes and survival of nonsmall cell lung cancer (NSCLC) by analyzing genotyping data from two published genome-wide association studies (GWASs). In the discovery, we used a genotyping dataset from the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial in the multivariable Cox proportional hazards regression analysis. We used Bayesian false discovery probability (≤0.80) for multiple testing correction to evaluate associations between 25,819 (2,176 genotyped and 23,643 imputed) single-nucleotide polymorphisms (SNPs) in 162 genes and survival of 1,185 NSCLC patients. Subsequently, we validated the identified significant SNPs with an additional 984 NSCLC patients from the Harvard Lung Cancer Susceptibility GWAS study. Finally, we found that three independent and potentially functional SNPs in three different genes (i.e., PLIN2 rs7867814 G>A, SULT2A1 rs2547235 C>T and UGT1A9 rs2011404 C>T) were independently associated with risk of death from NSCLC, with a combined hazards ratio of 1.22 [95% confidence interval = 1.09–1.36 and p = 0.0003], 0.82 (0.74–0.91 and p = 0.0002) and 1.21 (1.10–1.33 and p = 0.0001), respectively. Additional expression quantitative trait loci analysis found that the survival-associated PLIN2 rs7867814 GA + AA genotypes, but not the genotypes of other two SNPs, were significantly associated with increased mRNA expression levels (p = 0.005). These results indicated that PLIN2 variants may be potential predictors of NSCLC survival through regulating the PLIN2 expression.