Survivin promoter-regulated oncolytic adenovirus with Hsp70 gene exerts effective antitumor efficacy in gastric cancer immunotherapy

Gene therapy is a promising adjuvant therapeutic strategy for cancer treatment. To overcome the limitations of current gene therapy, such as poor transfection efficiency of vectors, low levels of transgene expression and lack of tumor targeting, the Survivin promoter was used to regulate the selective replication of oncolytic adenovirus in tumor cells, and the heat shock protein 70 (Hsp70) gene was loaded as the anticancer transgene to generate an AdSurp-Hsp70 viral therapy system. The efficacy of this targeted immunotherapy was examined in gastric cancer. The experiments showed that the oncolytic adenovirus can selectively replicate in and lyse the Survivin-positive gastric cancer cells, without significant toxicity to normal cells. AdSurp-Hsp70 reduced viability of cancer cells and inhibited tumor growth of gastric cancer xenografts in immuno-deficient and immuno-reconstruction mouse models. AdSurp-Hsp70 produced dual antitumor effects due to viral replication and high Hsp70 expression. This therapeutic system used the Survivin promoter-regulated oncolytic adenovirus vector to mediate targeted expression of the Hsp70 gene and ensure safety and efficacy for subsequent gene therapy programs against a variety of cancers.     

热休克蛋白70在肝细胞癌中的研究进展

热休克蛋白70(Heat shock protein 70,HSP70)是高度保守的多肽蛋白,作为分子伴侣,在蛋白稳态、细胞凋亡、侵袭、细胞信号转导中发挥重要的作用。研究表明HSP70的表达水平与肿瘤的发生发展及转归有密切联系,其在正常情况下呈低水平表达,而在肿瘤组织中呈高表达。肝细胞癌起病隐匿,相关研究报道HSP70表达水平的高低可以作为早期肝细胞癌诊断和鉴别诊断的敏感性生物学指标,同时影响着肝细胞癌患者的治疗和预后,本文就此做一综述。     

Genomic characterization of a large panel of patient-derived hepatocellular carcinoma xenograft tumor models for preclinical development

Lack of clinically relevant tumor models dramatically hampers development of effective therapies for hepatocellular carcinoma (HCC). Establishment of patient-derived xenograft (PDX) models that faithfully recapitulate the genetic and phenotypic features of HCC becomes important. In this study, we first established a cohort of 65 stable PDX models of HCC from corresponding Chinese patients. Then we showed that the histology and gene expression patterns of PDX models were highly consistent between xenografts and case-matched original tumors. Genetic alterations, including mutations and DNA copy number alterations (CNAs), of the xenografts correlated well with the published data of HCC patient specimens. Furthermore, differential responses to sorafenib, the standard-of-care agent, in randomly chosen xenografts were unveiled. Finally, in the models expressing high levels of FGFR1 gene according to the genomic data, FGFR1 inhibitor lenvatinib showed greater efficacy than sorafenib. Taken together, our data indicate that PDX models resemble histopathological and genomic characteristics of clinical HCC tumors, as well as recapitulate the differential responses of HCC patients to the standard-of-care treatment. Overall, this large collection of PDX models becomes a clinically relevant platform for drug screening, biomarker discovery and translational research in preclinical setting.     

Cabozantinib inhibits tumor growth and metastasis of a patient-derived xenograft model of papillary renal cell carcinoma with MET mutation

MET plays an important role in the development and progression of papillary renal cell carcinoma (pRCC). Evaluation of efficacy of MET inhibitors against pRCC has been hampered by limited preclinical models depicting MET abnormalities. We established a new patient-derived xenograft (PDX) model of pRCC carrying an activating mutation of MET and tested the ability of cabozantinib, an inhibitor of receptor tyrosine kinases including MET, to inhibit tumor growth and metastasis. Precision-cut, thin tissue slices from a pRCC specimen obtained by nephrectomy were implanted under the renal capsule of RAG2^?/?γC^?/? mice to establish first generation TSG-RCC-030. Histologic and genetic fidelity and metastatic potential of this model were characterized by immunohistochemistry, direct DNA sequencing and quantitative polymerase chain reaction (qPCR). The effect of cabozantinib on tumor growth and metastasis was evaluated. Whether measurements of circulating tumor DNA (ctDNA) by allele-specific qPCR could be used as a biomarker of tumor growth and response to therapy was determined. Subrenal and subcutaneous tumor grafts showed high take rates and metastasized to the lung. Both primary tumors and metastases expressed typical markers of pRCC and carried the same activating MET mutation as the parental tumor. Cabozantinib treatment caused striking tumor regression and inhibited lung metastasis in TSG-RCC-030. Plasma ctDNA levels correlated with tumor volume in control mice and changed in response to cabozantinib treatment. TSG-RCC-030 provides a realistic preclinical model to better understand the development and progression of pRCC with MET mutation and accelerate the development of new therapies for pRCC.     

人源细胞因子诱导的杀伤细胞联合化疗药物对裸鼠肝癌移植瘤的抑制作用

目的 比较单独应用细胞因子诱导的杀伤(CIK)细胞、化疗药物和CIK细胞联合化疗药物对肝癌裸鼠移植瘤生长的抑制作用,为临床上联合应用CIK细胞和化疗药物治疗肿瘤提供实验依据。方法 应用成分血分离机采集5例健康自愿者外周血单个核细胞(PBMC),经含IFN-γ、IL-2以及抗CD3单抗的细胞因子鸡尾酒诱导成CIK细胞,以流式细胞仪检测细胞表型。裸鼠肩胛部皮下接种BEL-7402肝癌细胞,第5天起相同部位分别给予生理盐水(对照组)、不同剂量的CIK细胞(CIK-1组和CIK-2组)、丝裂霉素-C(单纯化疗组)和CIK细胞联合丝裂霉素-C(联合治疗组),观察它们对肝癌生长的抑制作用。结果多因子诱导培养后,CD3、CD3^ CD8^ 、CD3^ CD56^ 和CD25^ 效应细胞亚群的比例明显升高,分别由最初的64.0％、28.0％、7.8％和9.1％,上升至94.7％、67.7％、61.3％和84.0％,其中CD3^ 、CD3^ CD8^ 细胞的比例在培养期间内可维持较高水平,CD25^ 和CD3^ CD56^ 细胞的比例分别于培养后的第7天和第13天达最高值,而后开始下降。在90d的观察期内,对照组、化疗组、CIK-1组、CIK-2组和联合治疗组的裸鼠成瘤牢分别为100％、70.0％、80.0％、70.0％和66.7％,生存率分别为10.0％、60.0％、40.0％、50.0％和75.0％,而且联合治疗组肿瘤生长缓慢,肿瘤组织坏死明显。结论 CIK细胞联合化疗药物对裸鼠肝癌移植瘤生长的抑制作用大于单独应用CIK细胞或化疗药物。     

CEA promoter-regulated oncolytic adenovirus-mediated Hsp70 expression in immune gene therapy for pancreatic cancer

Gene therapy is an important means for the comprehensive treatment of pancreatic cancer. Challenges associated with gene therapy include control of vector security and effective genetic screening. In this paper, a CEA promoter-regulated oncolytic adenovirus vector was constructed. The reporter gene assay demonstrated that the viral vector was confirmed to have tumor-specific replication features. In vitro cytology studies showed that the CEA promoter regulated the proliferation of the adenovirus vector carrying the Hsp70 gene (AdCEAp-Hsp70), which significantly increased the expression levels of Hsp70 in the CEA-positive pancreatic cancer cells, resulting in an overall reduction in the survival of cancer cells. In the human pancreatic cancer Panc-1 xenograft model in immune deficient nude mice, the CEA promoter-regulated adenovirus AdCEAp-Hsp70 significantly inhibited tumor growth. In the rat pancreatic cancer DSL-6A/C1 xenograft model in rats, the viral proliferation and high expression levels of Hsp70 promoted the interstitial infiltration of CD4+, CD8+ and gamma/delta T cells into tumors, induced host secretion of the cytokines TGF-β, INF-γ, and IL-6 and had a dual anti-tumor effects that completely inhibited the growth of pancreatic cancer. The results demonstrated that the oncolytic adenovirus under the control of CEA promoter provides additional assurances regarding the safety and efficiency of cancer gene therapy. This gene therapy model improves anti-cancer efficiency and has broad applications and developmental prospects.     

Targeted expression of BikDD combined with metronomic doxorubicin induces synergistic antitumor effect through Bax activation in hepatocellular carcinoma

Conventional chemotherapy is commonly used to treat advanced non-resectable hepatocellular carcinoma (HCC) but this treatment modality has not demonstrated convincing survival benefit in HCC patients. Our previous studies indicated that targeted expression of therapeutic BikDD driven by a liver cancer-specific α-fetoprotein promoter/enhancer (eAFP) in the VISA backbone (eAFP-VISA-BikDD) significantly and specifically kills HCC cells in multiple orthotopic animal models. To enhance its therapeutic efficacy, we combined eAFP-VISA-BikDD with chemotherapeutic agents and found that eAFP-VISA-BikDD plus doxorubicin (Dox) or 5-fluorouracil (5-FU) demonstrated synergistic cytotoxicity in HCC cells. Specifically, the combination of eAFP-VISA-BikDD plus Dox markedly induced apoptosis via increased Bax mitochondrial translocation and cytoplasmic cytochrome c release. Compared with either agent alone, a low dose of Dox combined with eAFP-VISA-BikDD induced better antitumor effect and prolonged longer survival of mice in two orthotopic liver cancer xenograft models. Our findings provide strong preclinical support for evaluating the combined therapy of eAFP-VISA-BikDD and Dox in a clinical setting as a treatment option for HCC.     

TACE联合索拉非尼治疗中晚期肝细胞性肝癌的临床研究

目的探讨经导管动脉化疗栓塞（transcatheterarterialchemoembolization,TACE）联合索拉非尼治疗中晚期肝细胞性肝癌（hepatocellularcarcinoma,HCC）的疗效及安全性。方法选择我院70例中晚期HCC患者,其中35例给予TACE联合索拉非尼治疗（观察组）,35例单纯行TACE治疗（对照组）。每4-8周根据实体瘤疗效评估标准（RECIST）行肿瘤应答评价,评估临床疗效及索拉非尼毒副反应,比较两组患者治疗后的中位生存期及中位疾病进展时间。结果观察组和对照组中位0s分别为14_8个月和8．2个月,差异有统计学意义（P〈0．05）,中位TIP分别为10．3个月和5．8个月,差异有统计学意义（P〈0．05）。观察组服用索拉非尼后有27例（77．1％）患者出现毒副反应,经对症治疗后好转。结论TACE联合索拉非尼治疗中晚期HCC疗效好,不良反应可耐受,有望成为中晚期HCC的一种治疗模式。     

An oncolytic adenovirus regulated by a radiation-inducible promoter selectively mediates hSulf-1 gene expression and mutually reinforces antitumor activity of I131-metuximab in hepatocellular carcinoma

Gene therapy and antibody approaches are crucial auxiliary strategies for hepatocellular carcinoma (HCC) treatment. Previously, we established a survivin promoter-regulated oncolytic adenovirus that has inhibitory effect on HCC growth. The human sulfatase-1 (hSulf-1) gene can suppress the growth factor signaling pathways, then inhibit the proliferation of cancer cells and enhance cellular sensitivity to radiotherapy and chemotherapy. I¹³¹-metuximab (I¹³¹-mab) is a monoclonal anti-HCC antibody that conjugated to I¹³¹ and specifically recognizes the HAb18G/CD147 antigen on HCC cells. To integrate the oncolytic adenovirus-based gene therapy and the I¹³¹-mab-based radioimmunotherapy, this study combined the CArG element of early growth response-l (Egr-l) gene with the survivin promoter to construct a radiation-inducible enhanced promoter, which was used to recombine a radiation-inducible oncolytic adenovirus as hSulf-1 gene vector. When I¹³¹-mab was incorporated into the treatment regimen, not only could the antibody produce radioimmunotherapeutic effect, but the I¹³¹ radiation was able to further boost adenoviral proliferation. We demonstrated that the CArG-enhanced survivin promoter markedly improved the proliferative activity of the oncolytic adenovirus in HCC cells, thereby augmenting hSulf-1 expression and inducing cancer cell apoptosis. This novel strategy that involved multiple, synergistic mechanisms, including oncolytic therapy, gene therapy and radioimmunotherapy, was demonstrated to exert an excellent anti-cancer outcome, which will be a promising approach in HCC treatment.     

Increased expression of inducible HSP70 in apoptotic cells is correlated with their efficacy for antitumor vaccine therapy

Apoptosis is a physiologic process in normal development, tissue remodeling and cell turnover. This cell death is noninflammatory and nonimmunogenic, but when associated with a danger signal, it can activate the immune system. However, the capacity of apoptotic cells to activate the immune system is not clearly established, although dead tumor cells have been largely exploited as a source of TAA in cellular therapy against cancer. From these cellular preparations, contradictory results have been reported on the effect of apoptotic cells as an effective source of TAA and their immunologic properties. These conflicting data strongly suggest that the optimal preparation of apoptotic cells derived from tumor cells remains to be determined. In this work, we studied and compared the efficacy of antitumor immune responses derived from repeated injections using different preparations of apoptotic cells. We investigated the importance of HSP70 and TGF-beta expression in apoptotic cells used in the treatment of an established and nonimmunogenic rat carcinoma. UVB-mediated apoptosis did not affect TGF-beta expression in tumor cells, whereas HS treatment sharply downregulated it. Thus, downregulation of TGF-beta permits normal DC activation and maturation and the induction of tumor immunity. We conclude that HS followed by UVB irradiation is a superior source of tumor antigen for the treatment of established tumors. Future work will determine whether HS independently upregulates HSP70, thereby suppressing expression of active TGF-beta, or whether the 2 are linked via a still undefined mechanism.     

HSP70ASODN转染增强顺铂抑制肝癌细胞HepG2增殖的试验研究

目的:研究HSP70ASODN联合顺铂对于肝癌细胞的增殖抑制作用。方法:运用MTT法比较转染组和未转染组联合顺铂在肝癌细胞增殖方面的差异。结果:观察转染HSP70组与未转染组HepG2细胞株对化疗药物顺铂的敏感性时,24h、48h、72h、96h未转染组与转染组的细胞生长抑制率分别为,0.329±0.029、0.583±0.022;0.356±0.035、0.684±0.052;0.497±0.033、0.723±0.048;0.523±0.035、0.715±0.047。结论:HSP70ASODN能增强顺铂对HepG2增殖抑制作用。     

热休克蛋白70mRNA反义寡核苷酸对荷瘤小鼠膀胱癌的作用

目的探讨热休克蛋白70（HSP70）mRNA反义寡核苷酸（ASO）对体内膀胱癌发生发展的影响及其可能的作用机制。方法用培养的人类膀胱癌细胞株BIU-87建立40只BALB／c裸鼠膀胱癌皮下荷瘤模型，待皮下肿瘤体积约为100mm^3时，随机分为4组，每组10只。HSP70 mRNA ASO加丝裂霉素C（MMC）处理组；HSP70 mRNA反义寡核苷酸组；丝裂霉素C组；空白对照组。HSP70 mRNA按10mg／kg，肿瘤局部注射，每周2次；MMC 0．1mg／kg，腹腔注射，每周2次，空白组用生理盐水代替上述处理。治疗的第30天托颈处死裸鼠，剥离出皮下肿瘤，照像并称重量。免疫组织化学法检测肿瘤组织中的微血管密度（MVD），RT—PCR和Western blotting检测HSP70表达；原位末端标记法（TUNEL）检测细胞凋亡。结果HSP70 mRNA ASO加MMC组肿瘤抑制率超过50％，高于HSP70 mRNA ASO组或MMC组，差异有统计学意义（P〈0．05）。HSP70 mRNA ASO和MMC组相比，差异无统计学意义（P〉0．05）。ASO＋MMC组的凋亡指数（AI）分别高于其他三组，差异有统计学意义（P〈0．05）；MMC和ASO组AI分别高于空白组（P〈0．05）；MMC和ASO组相比，差异无统计学意义（P〉0．05）。肿瘤组织中的MVD与上述结果一致。结论HSP70 mRNA ASO局部注射可以抑制肿瘤的生长，其作用机制可能和其抑制微血管的形成及促进细胞凋亡有关。     

FOXM1 expression predicts the prognosis in hepatocellular carcinoma patients after orthotopic liver transplantation combined with the Milan criteria

Molecular biomarker has been proposed to improve patient selection and post-transplant prognostication, but rare achievement has been made. In the present study, Forkhead box M1 (FOXM1) expression and its prognostic role have been investigated in hepatocellular carcinoma (HCC) treated by orthotopic liver transplantation (OLT). We found that the notably higher level of FOXM1 in tumors was associated with malignant pathological features of HCC and unfavorable outcome after OLT. The status of FOXM1 expression combined with the Milan criteria could make the prognostication more accurate and may be of particular interest for expanding the criteria in selecting transplant candidates.     

Successful treatment of multiple hepatocellular carcinoma with tumor thrombi in the major portal branches by intraarterial 5-fluorouracil perfusion chemotherapy combined with subcutaneous interferon-alpha and hepatectomy

We experienced a patient who received successful treatment for multiple hepatocellular carcinoma (HCC) nodules, with tumor thrombi in the major portal branches, with intraarterial 5-fluorouracil perfusion chemotherapy combined with subcutaneous interferon-alpha administration. The patient was a 50-year-old man with hepatitis C virus and HCC. The tumors consisted of a 5-cm main nodule in the right lobe (segment 8) and multiple intrahepatic metastases. The tumor also involved portal vein thrombosis throughout the right portal branch. After two cycles of interferon-alpha/5-fluorouracil combination chemotherapy, tumor markers demonstrated a decreasing tendency. Nine months after the initiation of this therapy, the tumors were limited to the right lobe and were surgically removed by S8 subsegmentectomy, S5 partial hepatectomy, and portal thrombectomy. The serum levels of both alpha-fetoprotein and protein induced by vitamin K absence II fell to normal levels after hepatic resection. Fifty-eight months after the first treatment, he is alive with several recurrent nodules in the liver. In conclusion, the interferon-alpha/5-fluorouracil combination therapy is a useful treatment for HCC in patients who have multiple intrahepatic metastases and portal vein thrombosis. In addition to this therapy, combined modality therapy including, for example, surgical resection, can sometimes have a dramatic therapeutic effect, shown by tumor markers reverting to normal levels.     

Combination treatment with hypoxia-activated prodrug evofosfamide (TH-302) and mTOR inhibitors results in enhanced antitumor efficacy in preclinical renal cell carcinoma models

Tumors often consist of hypoxic regions which are resistant to chemo- and radiotherapy. Evofosfamide (also known as TH-302), a 2-nitroimidazole triggered hypoxia-activated prodrug, preferentially releases the DNA cross-linker bromo-isophosphoramide mustard in hypoxic cells. The intracellular kinase mTOR plays a key role in multiple pathways which are important in cancer progression. Here we investigated the enhanced efficacy profile and possible mechanisms of evofosfamide in combination with mTOR inhibitor (mTORi) everolimus or temsirolimus in renal cell carcinoma (RCC) xenograft models. The antitumor activities of the mTORi everolimus or temsirolimus alone, evofosfamide alone, or the combination were investigated in the 786-O and Caki-1 RCC cells in vitro and in vivo xenograft models. Two schedules were tested in which evofosfamide was started on the same day as the mTORi or 1 week after. Combination mechanisms were investigated by measuring a panel of pharmacodynamic biomarkers by immunohistochemistry. Antitumor efficacy in both RCC xenograft models was enhanced by the combination of evofosfamide and mTORi. Evofosfamide reduced the increased hypoxia induced by mTORi. Combination treatment induced increased DNA damage, decreased cell proliferation, and decreased survivin. Addition of mTORi did not change evofosfamide-mediated cytotoxicity in 786-O or Caki-1 cells in vitro which might suggest cell non-autonomous effects, specifically increased tumor hypoxia, are important for the in vivo combination activity. Taken together, evofosfamide potentiates the antitumor efficacy of mTOR inhibitors and inhibits the increased tumor hypoxia caused by mTOR inhibition. These studies provide a translational rationale for combining evofosfamide with mTOR inhibitors in clinical studies.     

DLC-1基因在不同淋巴道转移潜能小鼠肝癌细胞系中表达的研究

目的:探讨DLC-1基因在小鼠肝癌淋巴道转移中的作用。方法:采用实时定量聚合酶链反应(RQ-PCR)方法检测DLC-1基因在淋巴道低转移潜能的小鼠肝癌细胞系Hca-P和高转移潜能的小鼠肝癌细胞系Hca-F中的表达水平。结果:Hca-P和Hca-F细胞系DLC-1基因表达分别为(3.83±0.45)×10-3和(2.02±0.13)×10-3,差异有统计学意义,P<0.01。随转移潜能的增高,DLC-1基因表达水平下降。免疫荧光细胞化学分析显示,DLC-1在低转移细胞株Hca-P中的荧光表达强度高于高转移细胞株Hca-F。结论:DLC-1基因作为转移抑制相关的基因,在肝癌的淋巴道侵袭转移中可能有着重要作用。DLC-1基因表达水平有可能成为预测肝癌转移的指标和基因治疗中新的靶点。     

基于列线图构建和验证肝细胞癌伴门静脉癌栓患者肝切除术后的预后评估模型

目的 分析影响肝细胞癌伴门静脉癌栓（PVTT-HCC）患者肝切除术后预后的影响因素,并基于列线图模型构建和验证预后评估模型。方法 本研究为回顾性队列研究,选择2008年1月—2017年11月在本院行肝切除术的PVTT-HCC患者为研究对象,随访截至2021年1月。主要预测结局为1、3、5年总生存率。按照7∶3的比例将患者随机分为训练集和验证集,在训练集中采用Cox比例风险回归分析影响预后的影响,并基于影响因素构建列线图模型。同时在训练集和验证集中采用C-index评价模型的区分度,一致性曲线评估模型的校准度。结果 共231例患者符合纳入排除标准纳入分析,其中训练集162例,验证集69例。Cox比例风险回归模型显示,AFP≥400 μg/L、AST≥40 U/L、ALP≥80 U/L、肿瘤个数>1个及肿瘤包膜不完整是影响预后的危险因素。在训练集中,列线图模型预测1、3、5年总生存率的C-index分别为0.826（95%CI: 0.791~0.861）、0.818（95%CI：0.782~0.854）、0.781（95%CI：0.742~0.820）,在验证集中分别为0.814（95%CI：0.777~0.851）、0.798（95%CI：0.758~0.837）、0.769（95%CI：0.728~0.810）。校正曲线显示列线图模型在训练集和验证集均有较好的校准度。结论 本研究构建的列线图模型可准确预测PVTT-HCC患者的预后。     

根治性肿瘤切除术联合胆管癌栓取出术治疗肝细胞癌合并胆管癌栓的疗效观察

目的 观察根治性肿瘤切除术联合胆管癌栓取出术治疗肝细胞癌合并胆管癌栓的疗效.方法 回顾性分析86例肝细胞癌合并胆管癌栓患者的临床资料,根据治疗方式的不同将患者分为观察组和对照组,每组各43例.观察组患者接受根治性肿瘤切除术联合胆管癌栓取出术,对照组患者接受肝叶(段)不规则性切除联合胆管癌栓取出术,比较两组患者的术后并发症发生率和生存时间.结果 观察组患者消化道出血、腹腔出血、腹腔积液和肝衰竭发生率高于对照组,膈下脓肿和胆汁漏发生率低于对照组,但差异均无统计学意义(P﹥0.05).观察组患者的累积生存率为51.2%,中位生存时间为27个月(95%CI:23.971~30.029);对照组患者的累积生存率为25.6%,中位生存时间为25个月(95%CI:22.253~27.747).观察组患者的累积生存率高于对照组,差异有统计学意义(χ2=6.581,P﹤0.05).结论 根治性肿瘤切除术联合胆管癌栓取出术对延长肝细胞癌合并胆管癌栓患者的生存期时间,提高患者生存率具有明显作用,但需要注意并发症预防,同时积极进行对症治疗.     

DC疫苗联合细胞因子诱导的杀伤细胞治疗转移性肾癌的长期疗效分析

[摘要] 目的：评价树突状细胞（DC）疫苗联合细胞因子诱导的杀伤（CIK）细胞治疗转移性肾癌的长期临床疗效及随访观察。方法：选择2011 年1 月至2013 年12 月于中国人民解放军总医院第五医学中心造血干细胞移植科接受DC疫苗联合CIK细胞治疗的29 例转移性肾癌患者,其中男性24 例、女性5 例,中位年龄55 岁（32～81 岁）。一线治疗12 例,分子靶向药物或细胞因子治疗进展后二线治疗6 例,三线及以上治疗11 例（包括二次手术治疗、细胞因子治疗和多线分子靶向药物治疗）。通过基因转染技术获得成熟的DC疫苗,体外细胞培养技术获得CIK细胞,经淋巴引流区及静脉输注方式按疗程回输患者体内,评价其长期临床疗效及总体生存率。结果：29 例患者中位随访时间5 年（1～7 年）。治疗疗程≥2（2～23 个疗程）。疗效评价：完全缓解1 例（3.4%）,部分缓解9 例(31%),疾病稳定13 例(44.8%),疾病进展6 例(20.7%)。客观反应率34.4%,疾病控制率79.2%。疾病稳定1年以上19 例（65.5%）。1、3、5 年生存率分别为93.1%(27/29)、65.5%（20/29）和51.7%（15/29）。中位无进展生存期及中位生存期均未达到。治疗期间均未观察到3 级以上不良反应发生。结论：DC疫苗联合CIK细胞治疗转移性肾细胞癌疗效肯定,可获得良好的疾病控制率及长期生存,安全性可控,延长了晚期肾癌患者的生存期。