B7-H4在卵巢癌中的研究进展

B7-H4分子又称B7S1和B7x，是最近发现的T细胞共刺激分子B7家族中的新成员，能负性调控T细胞的免疫应答、阻碍细胞因子的产生和细胞周期的进程。同时大量表达B7-H4分子可以保护表皮细胞免于失巢凋亡，促进上皮细胞的恶性转化，在恶性肿瘤的发生、进展和转归的免疫逃逸机制中发挥重要作用。B7-H4分子在质膜上的定位及其在卵巢癌组织和细胞上的过度表达使其具有非常诱人的应用前景，能潜在地作为卵巢癌早期诊断新的生物标志，并为临床上卵巢癌的靶向治疗提供了思路。     

PD-L1、B7-H3及B7-H4在宫颈癌免疫治疗中的研究进展

宫颈癌是我国常见的妇科恶性肿瘤,其发生和发展常与免疫逃逸有关,免疫治疗已经成为目前宫颈癌治疗的研究热点.随着免疫检查点程序性死亡配体1(PD-L1)及程序性死亡受体1(PD-1)的抑制剂在多种恶性肿瘤中的成功应用,以PD-L1为代表的共刺激因子与宫颈癌的研究愈发受到关注,其中负性共刺激因子与宫颈癌免疫抑制密切相关.PD...     

负性共刺激因子B7-H4在宫颈癌中的研究进展

宫颈癌是妇科最常见的恶性肿瘤,而恶性肿瘤的发生、发展常与免疫抑制有关。在肿瘤免疫中,T细胞免疫应答发挥关键作用。T细胞活化的第2条信号通路需要共刺激因子与其受体相结合。缺乏第二信号共刺激因子,第1条信号通路形成的抗原复合物会导致T细胞失活或功能紊乱。B7家族是近年针对共刺激因子的研究热点,可对T细胞免疫应答起正性和(或)负性调节作用。综述负性共刺激因子B7-H4在宫颈癌患者血液和组织中的表达及意义,探讨其在宫颈癌研究中存在的问题;简要介绍B7-H4在卵巢癌患者血液和组织中的表达及意义。同时对B7-H4提示预后、辅助诊断及免疫治疗等进行展望,旨在进一步阐明宫颈癌免疫抑制机制,进一步探索有效的免疫靶向治疗手段。     

B7-H4在子宫内膜异位症中的表达及意义

目的:研究子宫内膜异位症(EMs)患者在位、异位内膜组织中B7同源体4(B7-H4)的表达情况及血清sB7-H4的水平变化,探讨其在EMs发生、发展中的作用及意义.方法:免疫组化法检测43例卵巢子宫内膜异位组织、43例在位内膜组织及30例正常子宫内膜组织中B7-H4的表达情况;ELISA夹心法检测EMs患者血清中sB7-H4的表达水平.结果:B7-H4主要表达于子宫内膜细胞的胞浆和胞膜.异位内膜、在位内膜、正常内膜的B7-H4阳性率分别为62.8％、55.8％和30.0％,组间差异显著(x2=8.067,P=0.018);不同月经周期中B7-H4的表达无显著差异(P＞0.05).EMs患者血清中sB7-H4的表达水平显著高于对照组[(36.23±5.67)μg/L vs (31.24±4.56) μg/L,t=2.398,P＜0.05].结论:B7-H4蛋白的异常表达可能与EMs的发生、发展有关,其可能成为EMs诊断和治疗的一个新靶点.     

卵巢浆液性癌B7-H4的表达及临床病理意义

目的:探讨B7-H4在卵巢上皮性肿瘤浆液性癌中的表达及其临床病理意义。方法:用逆转录聚合酶链反应(RT-PCR)技术及免疫组织化学染色的方法检测6例良性卵巢上皮性肿瘤、10例交界性卵巢上皮性肿瘤、41例卵巢浆液性癌、5例内膜样癌、2例透明细胞癌和10例粘液性癌标本中B7-H4 mRNA及蛋白表达,并结合临床病理资料对其中41例卵巢浆液性癌中B7-H4蛋白的表达进行分析。结果:B7-H4 mRNA在74例卵巢上皮性肿瘤中均有表达,B7-H4蛋白在良性卵巢上皮性肿瘤、交界性卵巢上皮性肿瘤、恶性卵巢上皮性肿瘤中的表达阳性比例分别为2/6、6/10、52/58,恶性标本阳性率明显高于非恶性标本,差异有统计学意义(P<0.05);58例恶性卵巢上皮性癌标本中浆液性癌、内膜样癌、透明细胞癌、黏液性癌的B7-H4蛋白阳性比例分别为41/41、5/5、2/2、4/10,前三种病理类型B7-H4蛋白阳性率明显高于最后一种,且差异有统计学意义(P<0.05);41例卵巢浆液性癌标本中B7-H4蛋白阳性细胞比例在<10%,>10%~50%,>50~80%,>80~100%4组中的分布差异无显著性(P>0.05),且其蛋白表达与临床分期及病理分级有关(P<0.05),而与患者年龄、腹水细胞学、淋巴结转移无关(P>0.05)。结论:B7-H4在卵巢浆液性癌的高表达及其与临床分级分期的关系,提示其可能与肿瘤发生发展有关,可为卵巢恶性肿瘤诊断及治疗提供靶位点。     

卵巢癌细胞B7-H4基因高表达抑制CTL细胞杀伤作用的研究

目的:探讨卵巢癌细胞B7-H4基因高表达对靶向诱导的CTL细胞体外杀伤的影响。方法:RT-PCR法扩增编码基因,构建真核表达载体PEGFP-N1/B7-H4,脂质体介导将重组质粒PEGFP-N1/B7-H4导入SKOV3细胞中,筛选建立高表达细胞株。细胞免疫组织化检学检测B7-H4蛋白的表达。利用免疫磁珠分离法(MACS)分离纯化脐血CD34+细胞并在体外诱导分化为DC,用反复冻融法从卵巢癌细胞SKOV3中提取可溶性冻融抗原并负载DC诱导生成CTL。MTT法检测经抗原负载后激活的CTL细胞对SKOV3/B7-H4、SKOV3/neo和SKOV3细胞的杀伤作用。结果:成功构建了人B7-H4真核表达载体,SK-OV3/B7-H4细胞稳定表达B7-H4蛋白。CTL细胞对SKOV3/B7-H4细胞杀伤作用明显低于阴性对照组(20.12%±3.14%vs36.34%±4.53%,P<0.05)。结论:卵巢癌细胞B7-H4基因表达可抑制靶向诱导的CTL细胞毒性,可能与肿瘤细胞免疫逃逸有着密切的关系。     

参与卵巢癌免疫逃逸的免疫抑制分子研究进展

转化生长因子β(TGF-β)、血管内皮生长因子(VEGF)、白细胞介素10(IL-10)、环氧合酶(COX)、前列腺素(PG)E2等是卵巢癌细胞分泌的主要免疫抑制分子.这些分子可通过抑制抗原提呈细胞(APC)有效提呈肿瘤抗原,阻断T细胞、自然杀伤(NK)细胞和巨噬细胞等的抗瘤活性,促进肿瘤新生血管生成等机制,使肿瘤细胞得以逃避机体免疫监视而增殖和转移.这些免疫抑制分子在卵巢癌组织中的表达及其在患者体内的含量变化可望成为辅助疗效判断、观察预后的临床检测指标,为临床最佳治疗方案的选择提供理论依据,并可作为开发抗瘤药物的新靶点,为卵巢癌的生物治疗提供新思路.     

谷氨酰胺代谢与卵巢癌相关研究进展

卵巢癌是妇科常见恶性肿瘤,由于卵巢解剖位置特殊,早期无特异性症状和体征,超过70%的患者确诊已属晚期,加之化疗耐药,卵巢癌患者往往死亡率高,预后差。代谢重编程是恶性肿瘤的最重要特征之一。最新研究表明,谷氨酰胺代谢参与卵巢癌的恶性增殖、侵袭和转移,并与卵巢癌化疗耐药及免疫逃逸等密切相关,是靶向卵巢癌能量代谢的重要策略之一。本文综述了谷氨酰胺代谢参与卵巢癌的恶性进展以及化疗耐药、免疫逃逸等相关研究进展。     

细胞周期蛋白B1在卵巢癌中的研究进展

细胞周期蛋白B1(cyclin B1)是细胞周期G2/M期的重要调控因子,主要促进细胞有丝分裂和卵母细胞的成熟.生理情况下,eyclin B1在S期合成,G2期达高峰,M期进入胞核促进细胞分裂增殖.许多研究显示,eyelin B1含量的增加通常出现在永生化的肿瘤细胞,cyclin B1参与卵巢细胞网络调控系统,其改变与卵巢癌的发生及恶性生物学行为密切相关.cyclin B1蛋白在卵巢癌细胞周期中异常表达,呈现出癌基因的特性,cyclln B1过表达与定位的改变受多个基因的调节,而且与卵巢癌的发生发展、预后、诊断和治疗密切相关.     

免疫检查点抑制剂在卵巢癌治疗中的研究进展

免疫检查点在肿瘤免疫逃逸中起着至关重要的作用。细胞毒T淋巴细胞相关抗原4(CTLA-4)、程序性死亡受体-1(PD-1)及其配体PD-L1是近年研究比较透彻的免疫检查点分子。多项临床试验证实,CTLA-4和PD-1/PD-L1这些免疫检查点的抑制剂在控制恶性肿瘤生长、维持疾病稳定及延长患者生存时间上取得了令人瞩目的效果。卵巢癌是免疫原性肿瘤,将标准治疗与免疫治疗联合有望进一步改善患者预后。本文对免疫检查点分子在卵巢癌的表达、免疫检查点抑制剂在卵巢癌治疗中的进展及临床试验结果进行综述。     

B7-H4 overexpression in ovarian tumors

OBJECTIVES: Despite great advances in therapeutic management, the mortality rate for ovarian cancer has remained relatively stable over the past 50 years. This study was designed to evaluate the expression of B7-H4 protein, recently identified as a potential molecular marker of breast and ovarian cancer by quantitative PCR analysis, in benign tumors, tumors of low malignant potential and malignant tumors of the ovary. METHODS: Archival formalin-fixed tissue blocks from serous, mucinous, endometrioid and clear cell ovarian tumors were evaluated by immunohistochemistry for the distribution of B7-H4 expression, and staining intensity was measured by automated image analysis. Univariate analyses were used to test for statistically significant relationships. RESULTS: B7-H4 cytoplasmic and membranous expression was detected in all primary serous (n = 32), endometrioid (n = 12), and clear cell carcinomas (n = 15), and in all metastatic serous (n = 23) and endometrioid (n = 7) ovarian carcinomas. By contrast, focal B7-H4 expression was detected in only 1/11 mucinous carcinomas. The proportion of positive cells and median staining intensity was greater in serous carcinomas than in serous cystadenomas or serous tumors of low malignant potential, and the differences were statistically significant (P < 0.0001 and P = 0.034, respectively). The median staining intensity was also significantly greater in endometrioid carcinomas than in endometriosis (P = 0.005). CONCLUSIONS: The consistent overexpression of B7-H4 in serous, endometrioid and clear cell ovarian carcinomas and the relative absence of expression in most normal somatic tissues indicates that B7-H4 should be further investigated as a potential diagnostic marker or therapeutic target for ovarian cancer.     

A review of B7-H3 and B7-H4 immune molecules and their role in ovarian cancer

A number of members of the B7 superfamily of ligands have been implicated in tumor immunogenicity and cancer development. Two of these recently characterized ligands, B7-H4 and B7-H3, have been linked to ovarian tumors. B7-H4 is consistently overexpressed in ovarian tumor specimens, and its tissue and serum levels have been found to be a potential biomarker for ovarian cancer, either alone or in combination with CA125. More recently, B7-H3 has been found to be overexpressed in a large series of ovarian cancer tumor specimens and similar to other types of carcinomas, B7-H3 overexpression has been correlated with poor survival. On the basis of the results obtained by knocking down B7-H3 protein using siRNA, researchers have suggested that blocking the action of B7-H3 could reduce tumor growth, metastatic potential, and improve survival. Because siRNA knock-down is not an ideal clinical therapeutic vehicle, additional studies using antibody-mediated suppression of the B7-H3 protein are necessary to fully evaluate the clinical potential of this molecule as a therapeutic target.     

B7-H4 is over-expressed in early-stage ovarian cancer and is independent of CA125 expression

OBJECTIVE: This study characterizes the expression of the novel biomarker B7-H4 in ovarian cancer tissue, normal ovaries, and benign ovarian tumors, and evaluates its relationship to CA125. METHODS: Ovarian tissue lysates from 251 patients with ovarian carcinoma were assessed for the levels of B7-H4 and CA125 by ELISA assays. For comparison, ovarian tissues from patients with benign ovarian tumors (n=43) and patients with normal ovaries (n=32) were tested. The marker concentrations were correlated with CA125 expression, clinicopathological variables, and patient outcome. RESULTS: Using a cut-off based on the 95th percentile of B7-H4 or CA125 concentration in the control group, B7-H4 was over-expressed in 48% of patients with stage I cancer, 55% of patients with stage II cancer, and 67% of patients with late stage cancer. CA125 was elevated in 31% patients with early stage cancer. B7-H4 was elevated in tumors of 30 patients with early stage cancer that were negative for CA125. The combination of B7-H4 and CA125 identified 56 early stage cancer patients (65%) as positive. Correlation of marker expression to clinical outcome showed that high B7-H4 levels were correlated with poor prognosis. However, the effect was not significant when outcome was adjusted for other clinicopathological variables. CONCLUSION: B7-H4 expression was low in normal ovaries and in benign tumors while half of early stage and two-thirds of late stage cancers over-expressed B7-H4. The data are consistent with previous observations and support further investigation of B7-H4 in the detection of early stage ovarian cancer either alone, or in combination with CA125.     

Monoclonal antibody-based immunotherapy of ovarian cancer: Targeting ovarian cancer cells with the B7-H3-specific mAb 376.96

Objective

The high rate of relapse in patients with advanced ovarian cancer likely reflects the chemoresistance of cancer initiating cells (CICs). We evaluated the anti-tumor activity of monoclonal antibody (mAb) 376.96, which recognizes a B7-H3 epitope expressed on ovarian carcinoma cells (OCCs), in combination with the tyrosine kinase inhibitor Sunitinib and chemotherapy on chemosensitive and chemoresistant cells and CICs.

Methods

Eight ovarian cancer cell lines including platinum- and taxane-resistant cell lines were analyzed by flow cytometry to establish expression of the mAb 376.96-defined-B7-H3-epitope on differentiated ovarian cancer cells and CICs. Samples from 10 ovarian cancer patients were analyzed via immunohistochemistry for mAb 376.96-defined-B7-H3-epitope expression. In vitro studies assessed mAb 376.96 alone and in combination with Sunitinib on the growth of chemosensitive and chemoresistant cell lines and on the content of CICs.

Results

The mAb-376.96-defined-B7-H3 epitope is expressed on both differentiated cells and CICs in chemosensitive and chemoresistant ovarian cancer cell lines and 10 patient derived ovarian cancer tumors. In vitro treatment of chemoresistant cell lines with mAb 376.96 resulted in decreased cell viability. mAb 376.96 enhanced the cytotoxicity of Sunitinib and reduced the content of CICs.

Conclusion

The mAb-376.96-defined-B7-H3-epitope was found to be expressed on both differentiated ovarian cancer cells and CICs in chemosensitive and chemoresistant ovarian cancer cell lines. mAb 376.96 inhibited the in vitro growth of chemosensitive and chemoresistant OCCs and reduced the content of CICs when used with Sunitinib. Further studies examining B7-H3 as a potential target of mAb-based immunotherapy for this type of malignancy are warranted.     

主动循环热灌注化疗对卵巢癌腹水疗效及HE4、CA125、VEGF和B7-H4水平的影响

目的 探讨卵巢癌腹水患者采用主动循环腹腔热灌注化疗治疗的效果及对血清肿瘤相关标记物水平的影响.方法 选取84例卵巢癌伴腹水患者,根据腹腔热灌注化疗方法分为主动循环组和常规组各42例,两组患者采用紫杉醇+顺铂实施不同的腹腔热灌注治疗.结果 主动循环组与常规组进入腹腔时的灌注液体温度差异无统计学意义(P>0.05),抽出腹...