DC-CIK联合化疗治疗晚期非小细胞肺癌的临床疗效

目的:研究DC-CIK联合化疗治疗晚期非小细胞肺癌的疗效及安全性。方法:选取2008年8月至2010年1月就诊于山西省肿瘤医院的50例Ⅲ~Ⅳ晚期非小细胞肺癌患者,采用DC-CIK联合化疗[多西他赛(docetaxel)+顺铂(cisplatin)]为联合治疗组;选取临床资料相近的同期进行单纯化疗(多西他赛+顺铂)的50例Ⅲ~Ⅳ晚期非小细胞肺癌患者为单纯化疗组,比较两组患者治疗后的免疫功能、近期疗效、1年生存率、生活质量,并观察DC-CIK细胞治疗的安全性。结果:成功培养患者的DC-CIK细胞,其中的CD3+CD8+、CD3+CD56+细胞比例较培养前显著提高(P<0.05)。联合治疗组患者治疗后外周血各T细胞亚群均无明显变化,IFN-γ水平显著升高(P<0.05);单纯化疗组患者治疗后外周血CD3+CD4+、CD3+CD8+、CD3-CD56+细胞比例下降(P<0.05),IL-2、TNF-α水平明显降低(P<0.05)。联合治疗组患者的DCR为78.0%,显著高于单纯化疗组的56.0%(P<0.05);联合治疗组患者1年生存率为50.0%,与单纯化疗组44.0%的差别无统计学意义(P>0.05)。联合治疗组患者的不良反应(包括骨髓抑制、恶心呕吐、周围神经毒性)明显轻于单纯化疗组(P<0.05),联合治疗组患者治疗后体力、食欲较单纯化疗组改善明显。结论:与单纯化疗相比,DC-CIK联合化疗治疗晚期非小细胞肺癌安全、有效,可以提高缓解率,延长生存期,改善患者的生活质量。     

DC - CIK 治疗 43 例晚期非小细胞肺癌的近期疗效观察简

目的：探讨树突状细胞（dendritic cell,DC）共培养细胞因子诱导的杀伤细胞（cytokine-induced killer,CIK）治疗晚期非小细胞肺癌（non-small cell lung cancer,NSCLC）患者的近期疗效。方法：将43例确诊的晚期非小细胞肺癌患者给予DC-CIK治疗,观察治疗前后外周血中 T细胞亚群及细胞因子的变化,卡氏评分（Karnofsky,KPS）和临床疗效,并观察其不良反应。结果：患者治疗后CD3＋、CD4＋、CD56＋和CD4＋/CD8＋的比例较治疗前增加（P＜0.05）,差异有统计学意义。免疫治疗增加了细胞因子IL-2、IL-12、IFN-γ和TNF-α的水平（P＜0.05）。治疗后疾病控制率为53.49%,KPS评分总提高率为83.72%。结论：DC-CIK治疗能提高患者的免疫功能及生活质量,有望成为非小细胞肺癌有效的过继免疫治疗方法。     

晚期非小细胞肺癌患者DC-CIK治疗前的免疫状态与预后的关系

目的探讨晚期非小细胞肺癌(NSCLC)患者树突状细胞-细胞因子诱导的杀伤细胞(dendritic cellcytokine induced killers,DC-CIK)治疗前外周血CD4+CD25+CD127-调节性T细胞(regulatory T cells,TRegs)、CD3+CD56+细胞因子诱导的杀伤细胞(cytokine induced killer cells,CIKs)、CD4+/CD8+T细胞比值与预后的关系。方法采用流式细胞仪技术检测45例晚期NSCLC患者DC-CIK治疗前、后外周血CD4+CD25+CD127-TRegs、CD3+CD56+CIKs、CD4+和CD8+T细胞,并与患者临床特征进行相关分析;采用Cox回归模型分析治疗前CD4+CD25+CD127-TRegs、CD3+CD56+CIKs、CD4+/CD8+T细胞比值对患者预后的影响。结果晚期NSCLC患者DC-CIK治疗前、后比较,治疗后患者外周血CD4+T细胞减少,CD8+T细胞增多,CD4+/CD8+T细胞比值下降,差异均有统计学意义(均P<0.05);原发病灶瘤体最长直径越大,患者治疗前外周血TRegs越多,CIKs越少(均P<0.05);生存分析显示,治疗前TRegs越多,患者预后越差,生存期越短,且TRegs>7×109/L影响患者的生存(P<0.05)。结论晚期NSCLC患者外周血TRegs是判断预后的独立预测指标。     

DC-CIK细胞过继免疫治疗联合XELOX方案化疗对进展期结直肠癌患者的疗效分析

目的探讨树突状细胞-细胞因子诱导杀伤细胞(DC-CIK)细胞过继免疫治疗联合奥沙利铂+卡培他滨(XELOX)方案化疗对进展期结直肠癌(CRC)患者的疗效。方法将104例进展期CRC患者按随机数字表法分为观察组和对照组各52例,观察组行DC-CIK联合化疗,对照组行化疗。检测患者治疗前后血清糖类抗原242(CA242)、糖类抗原19-9(CA19-9)、癌胚抗原(CEA)、T淋巴细胞亚群[自然杀伤性T(NKT)细胞、自然杀伤(NK)细胞、CD4^+、CD3^+、CD8^+]及白细胞介素-6(IL-6)、白细胞介素-2(IL-2)、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)水平,评价患者近期疗效。结果观察组患者的缓解率(RR)为61.54%(32/52),疾病控制率(DCR)为75.00%(39/52),分别高于对照组的40.38%(21/52)和55.77%(29/52),差异均有统计学意义(P<0.05)。治疗后,两组患者血清CEA水平均低于治疗前,差异均有统计学意义(P<0.05);治疗后,观察组患者血清IL-2、IFN-γ、TNF-α、CD4^+、CD3^+、CD8^+及NKT细胞水平均高于对照组,差异均有统计学意义(P<0.05)。结论 CRC患者行DC-CIK细胞过继免疫治疗联合XELOX方案化疗可改善机体免疫功能,弥补单纯化疗所造成的免疫抑制,为安全有效的疗法。     

中晚期非小细胞肺癌同步放化疗与树突状细胞联合细胞因子活化杀伤细胞生物治疗的临床效果

目的探讨同步放化疗与树突状细胞-细胞因子活化杀伤细胞(DC-CIK)生物治疗在中晚期非小细胞肺癌(NSCLC)患者中的临床效果。方法 86例中晚期NSCLC患者随机分为观察组与对照组,每组43例,观察组给予DC-CIK与同步放化疗治疗,对照组给予单纯同步放化疗治疗,对比两组患者的临床疗效。结果观察组患者治疗有效率(RR)为90.7%(39/43),对照组为81.4%(35/43),差异有统计学意义(P>0.05)。观察组患者治疗后CD3~+、CD4~+/CD8~+、自然杀伤细胞(NK)水平均显著高于治疗前(P<0.01),对照组均显著低于治疗前(P<0.01)。观察组患者治疗后的CD3~+、CD4~+/CD8~+、NK水平显著高于对照组(P<0.01);观察组患者发热发生率显著高于对照组(P<0.01),粒细胞减少、放射性肺炎发生率显著低于对照组(P<0.05),两组患者胃肠道反应以及放射性食管炎发生率的差异无统计学意义(P>0.05)。结论 DC-CIK与同步放化疗联合用于中晚期NSCLC患者的治疗,可在获得与同步放化疗相同的近期疗效基础上,有效提高机体的抗肿瘤能力,减少粒细胞减少及放射性损伤等不良反应,值得临床推广。     

多西紫杉醇联合GP化疗方案对晚期非小细胞肺癌患者炎性因子、免疫功能及临床症状的影响

目的 探讨多西紫杉醇联合吉西他滨+顺铂(GP)化疗方案对晚期非小细胞肺癌患者炎性因子、免疫功能及临床症状的影响.方法 回顾性分析40例晚期非小细胞肺癌患者的临床资料,所有患者均接受多西紫杉醇联合GP化疗方案,化疗2个周期后,比较化疗前后患者的血清炎性因子、免疫功能及临床症状.结果 化疗2个周期后,患者的血清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、C-反应蛋白(CRP)浓度明显低于化疗前(P﹤0.01);CD3+、CD4+、CD4/CD8水平高于化疗前,CD8+水平低于化疗前,差异均有统计学意义(P﹤0.05);癌因性疲乏、局部疼痛及食欲不振评分均明显低于化疗前(P﹤0.01).结论 多西紫杉醇联合GP化疗方案能有效缓解晚期非小细胞肺癌患者的临床症状,可能与降低血清炎性因子水平、改善免疫功能等因素有关.     

晚期非小细胞肺癌患者外周血CD4~+ CD25~+ FOXP3~+调节性T细胞的表达及临床意义

目的探讨晚期非小细胞肺癌患者外周血CD4~+ CD25~+ FOXP3~+ 调节性T(Treg)细胞的表达及其临床意义。方法采用免疫荧光术及流式细胞仪检测50例晚期非小细胞肺癌患者及50例健康对照组外周血中CD4~+ CD25~+ FOXP3~+ Treg细胞、CD4~+ CD25~+ Treg细胞、CD4~+ T细胞和CD4~+ CTLA-4~+ T细胞的表达。结果晚期非小细胞肺癌患者外周血中CD4~+ CD25~+ FOXP3~+ Treg细胞、CD4~+ CD25~+ Treg细胞和CD4~+ CTLA-4~+ T细胞的比例均高于健康对照组(均P<0.05),CD4~+ T细胞的比例均低于健康对照组(均P<0.05)。结论晚期非小细胞肺癌患者外周血CD4~+ CD25~+ FOXP3~+ Treg细胞比例高于健康对照者,可能与肺癌患者的免疫抑制和肿瘤进展相关。     

DC-CIK联合放化疗对小细胞肺癌患者免疫功能的影响

目的：探讨细胞因子诱导的杀伤细胞（CIK）、树突状细胞（DC）免疫治疗联合放化疗对小细胞肺癌患者外周血淋巴细胞亚群的影响及疗效。方法收集60例小细胞肺癌患者资料,其中32例患者采用DC-CIK联合放化疗（化疗方案为依托泊苷+顺铂/依托泊苷+卡铂/伊立替康+顺铂,放疗方案为适形调强放疗技术）为联合治疗组;28例进行放化疗的患者为对照组。治疗结束2周后采用流式细胞仪检测外周血CD3+、CD3+CD4+、CD3+CD8+、NK细胞、Treg细胞及相关因子IFN-γ、IL-2、IL-10、TGF-β1的变化,并评价疗效。结果联合治疗组IFN-γ治疗后较治疗前升高,IL-10、TGF-β1较治疗前降低（P﹤0.05）;联合治疗组患者外周血CD3+CD8+、CD3+CD4+、NK细胞治疗后水平高于对照组（P﹤0.05）;而Treg细胞、IL-10、TGF-β1治疗后水平低于对照组（P﹤0.05）。对照组患者外周血CD3+、CD3+CD4+、CD3+CD8+、NK细胞治疗后较治疗前下降,差异有统计学意义（P﹤0.05）;细胞因子在治疗前后无变化。虽然联合组疾病控制率较对照组高,但差异无统计学意义（87.50%vs 71.43%,P﹥0.05）。结论自体DC-CIK治疗可以改善小细胞肺癌联合放化疗患者的细胞免疫功能,是否能改善生存需进一步研究。     

转移性肾癌自体DC-CIK细胞免疫治疗疗效观察

目的:观察自体树突状细胞（dendritic cell,DC）激活的细胞因子诱导的杀伤（cytokine-induced killer,CIK）细胞治疗转移性肾癌的临床效果及安全性。方法:32例转移性肾癌患者,采集外周血单个核细胞（peripheral blood mononuclear cell,PBMC）,体外制备成DC-CIK细胞,12天后连续三天回输,输注结束后次日开始下一个疗程,共计3个疗程。观察治疗前、治疗后6个月外周血中T细胞亚群及细胞因子IL-2、IFN-γ、IL-4及IL-10的变化,并统计治疗3年后生存率以及生活质量评分（KPS）,观察不良反应。结果:治疗前后比较CD4+无显著差异,CD8+显著降低,CD3+、CD56+以及CD4+/CD8+有显著差异（P<0.05）。两组治疗后6个月 IL-2、IFN-γ与治疗前比较均升高（P<0.05）。IL-4、IL-10与治疗前比较均降低（P<0.05）。治疗前、治疗后2周患者WBC、ALT、AST以及BUN、Cr比较无显著性差别（P>0.05）。3年生存率90.63%,治疗后1年生活质量评分平均升高24.7,显著高于治疗前的11.8(P<0.05)。不良反应轻微。结论:自体DC-CIK细胞输注后可显著提升转移性肾癌（RCC）患者免疫水平,安全性良好,可作为晚期RCC患者可选择的治疗方法。     

力尔凡联合NP方案治疗非小细胞肺癌疗效观察

目的 评价力尔凡在晚期非小细胞肺癌化疗中的临床应用价值.方法 晚期非小细胞肺癌60例,随机分为治疗组(NP方案加力尔凡)和对照组(单用NP方案).分别对患者的临床疗效、外周血T细胞哑群CD3+、CD4+、CD8+、CD4+/CD8+、LTT、外周WBC、毒副反应等指标进行比较.结果 近期疗效显示,加用力尔凡的治疗组中缓解率(RR)为63.3%,对照组RR为43.3%(P<0.01).化疗组与对照组比较,T细胞亚群CD3+、CD4+、CD4+/CD8+、LTT均有明显提高,CD8+降低,差异有统计学意义.治疗组外周WBC减少病例少于对照组,并且程度较轻.治疗组患者发生恶心、呕吐、神经毒性、静脉炎、脱发的毒副反应均有所减少,但发热病例增多.结论 力尔凡与化疗联合应用能提高晚期非小细胞肺癌的免疫功能,对化疗所致的WBC减少有保护作用,能减轻化疗引起的多种毒副反应.     

桥接整合因子1通过AKT-mTOR通路诱导非小细胞肺癌H1975细胞周期阻滞

目的：研究桥接整合因子1（bridging intergrator 1,Bin1）基因过表达后对非小细胞肺癌细胞株H1975细胞周期的影响及其作用机制。方法：构建携带Bin1基因的CMV-MCS-GFP-SV40-Neomycin-Bin1质粒,并转染H1975细胞（Bin1+组）,另设置空白质粒转染组（Bin1-组）及空白对照组（Ctrl组）,利用RT-PCR和Western blotting分别检测3组细胞中Bin1在mRNA和蛋白质水平的表达情况。流式细胞术检测不同处理组H1975细胞周期的变化,Western boltting分别检测各组中AKT、mTOR磷酸化水平及细胞周期相关蛋白（周期蛋白D1、CDK4、Rb）的表达情况。结果：与Bin1-组、Ctrl组比较,Bin1+组H1975细胞中Bin1在mRNA、蛋白水平表达明显上调（均P<0.05）; H1975细胞阻滞在G1期\[（60.53±1.89）% vs（46.14±1.56）%、（47.33±2.07）%,均P<0.05\]; Bin1+组H1975细胞内p-AKT、p-mTOR表达下调（均P<0.05）,AKT、mTOR表达变化无统计学差异（P>0.05）;周期蛋白D1、CDK4的表达量均明显下调(P<0.05),Rb表达量明显增加（P<0.05）。结论：Bin1基因在H1975细胞株过表达后明显诱导细胞周期阻滞,其机制可能是通过抑制AKT-mTOR通路及其细胞周期相关蛋白实现的。     

Radiation sensitivity of merkel cell carcinoma cell lines

: Merkel cell carcinoma (MCC), being a small cell carcinoma, would be expected to be sensitive to radiation. Clinical analysis of patients at our center, especially those with macroscopic disease, would suggest the response is quite variable. We have recently established a number of MCC cell lines from patients prior to radiotherapy, and for the first time are in a position to determine their sensitivity under controlled conditions.     

Mechanisms of kaposis-sarcoma cell supernatant-induced vascular cell invasion

Kaposi's sarcoma (KS) is a highly angiogenic lesion frequently associated with acquired immune deficiency syndrome. Histologically the lesions appear to contain proliferative 'spindle shaped' cells with a mixed smooth muscle-endothelial-fibroblastic histotype and a conspicuous neovascularization, derived from host cell recruitment. Media conditioned by cultured KS cells (KS-CM) have angiogenic properties. KS-CM is able to promote endothelial and smooth muscle cell migration and invasion. The mechanisms of this KS-CM activity are still unknown. We hypothesize that KS-CM contains numerous factors with different roles in inducing the neo angiogenic process. We show that AIDS-IST-KS cell supernatants induce gelatinase A production and plasminogen activator (PA) up-regulation in vascular cells. KS-CM activity in vivo is heparin dependent. Also bFGF alone, a heparin dependent factor, alone can induce endothelial and smooth muscle cell invasion, MMP-2 production and PA activity. However, antibodies to bFGF do not block KS-CM activity and do not reduce the effect on PA up-regulation. This evidence suggests that heparin-binding factors other than bFGF may be present. Chromatography of KS-CM on heparin-sepharose demonstrates the presence of two heparin-binding fractions with chemotactic and gelatinase A inducing activity. The flow through was also active. KS-CM absorption on heparin-sepharose beads did not modify its induction of PA activity, further evidence for the presence of non heparin-binding factors as well.     

NDRG2 suppresses the proliferation of clear cell renal cell carcinoma cell A-498

Background

Recently, the anti-tumor activity of N-myc downstream-regulated gene 2 (NDRG2) was shown decreased expression in clear cell renal cell carcinoma (CCRCC), but the role of the down-expression of NDRG2 has not been described.

Methods

The NDRG2 recombinant adenovirus plasmid was constructed. The proliferation rate and NDRG2 expression of cell infected with recombinant plasmid were mesured by MTT, Flow cytometry analysis and western blot.

Results

The CCRCC cell A-498 re-expressed NDRG2 when infected by NDRG2 recombinant adenovirus and significantly decreased the proliferation rate. Fluorescence activated cell sorter analysis showed that 25.00% of cells expressed NDRG2 were in S-phase compared to 40.67% of control cells, whereas 62.08% of cells expressed NDRG2 were in G1-phase compared to 54.39% of control cells (P < 0.05). In addition, there were much more apoptotic cells in NDRG2-expressing cells than in the controls (P < 0.05). Moreover, upregulation of NDRG2 protein was associated with a reduction in cyclin D1, cyclin E, whereas cyclinD2, cyclinD3 and cdk2 were not affected examined by western blot. Furthermore, we found that p53 could upregulate NDRG2 expression in A-498 cell.

Conclusions

We found that NDRG2 can inhibit the proliferation of the renal carcinoma cells and induce arrest at G1 phase. p53 can up-regulate the expression of NDRG2. Our results showed that NDRG2 may function as a tumor suppressor in CCRCC.     

Inhibition of S-adenosylhomocysteine hydrolase decreases cell mobility and cell proliferation through cell cycle arrest

S-adenosylhomocysteine hydrolase (AHCY) hydrolyzes S-adenosylhomocysteine to adenosine and l-homocysteine, and it is already known that inhibition of AHCY decreased cell proliferation by G2/M arrest in MCF7 cells. However, the previous study has not indicated what mechanism the cell cycle arrest is induced by. In this study, we aimed to investigate the different cell cycle mechanisms in both p53 wild-typed MCF7 and p53 mutant-typed MCF7-ADR by suppressing AHCY. We extensively proved that AHCY knockdown has an anti-proliferative effect by using the WST-1 assay, BrdU assay, and cell cytometry analysis and an anti-invasive, migration effect by wound-healing assay and trans-well analysis. Our study showed that down-regulation of AHCY effectively suppressed cell proliferation by regulating the MEK/ERK signaling pathway and through cell cycle arrests. The cell cycle arrest occurred at the G2/M checkpoint by inhibiting degradation of cyclinB1 and phosphorylation of CDC2 in MCF7 cells and at the G1 phase by inhibiting cyclinD1 and CDK6 in MCF7-ADR cells. Finally, we determined that AHCY regulates the expression of ATM kinase that phosphorylates p53 and affects to arrest of G2/M phase in MCF7 cells. The findings of this study significantly suggest that AHCY is an important regulator of cell proliferation through different mechanism in between MCF7 and MCF7-ADR cells as p53 status.     

Landscape of immune cell infiltration in clear cell renal cell carcinoma to aid immunotherapy

The tumor microenvironment, comprised of tumor cells and tumor-infiltrating immune cells, is closely associated with the clinical outcome of clear cell renal cell carcinoma (ccRCC) patients. However, the landscape of immune infiltration in ccRCC has not been fully elucidated. Herein, we applied multiple computational methods and various datasets to reveal the immune infiltrative landscape of ccRCC patients. The tumor immune infiltration (TII) levels of 525 ccRCC patients using a single-sample gene were examined and further categorized into immune infiltration subgroups. The TII score was characterized by distinct clinical traits and showed a significant divergence based on gender, grade, and stage. A high TII score was associated with the ERBB signaling pathway, the TGF-β signaling pathway, and the MTOR signaling pathway, as well as a better prognosis. Furthermore, patients with high TII scores exhibited greater sensitivity to pazopanib. The low TII score was characterized by a high immune infiltration level of CD8⁺ T cells, T follicular helper cells, and regulatory T cells (Tregs). Moreover, the immune check point genes, including CTLA-4, LAG3, PD-1, and IDO1, presented a high expression level in the low TII score group. Patients in the high TII score group demonstrated significant therapeutic advantages and clinical benefits. The findings in this study have the potential to assist in the strategic design of immunotherapeutic treatments for ccRCC.     

Characterisation of four merkel cell carcinoma adherent cell lines

We have previously described the establishment of a number of cell lines from Merkel cell carcinoma (MCC), also known as small cell cancer of the skin or neuroendocrine carcinoma of the skin. These cells, all of which grew as suspension cultures, were found to resemble small cell lung cancer (SCLC) lines types 1, 2 and 3 by their morphology and growth characteristics. We now report 4 more MCC cell lines which resemble the SCLC type 4 cell lines in that they grow as adherent monolayers. These MCC lines would belong to the variant subgroup as they no longer express most neuroendocrine markers, grow at low cell density and have population doubling times of 1–5 days in contrast to the MCC suspension lines which have doubling times of 6–12 days. MCC 14/1 and MCC 14/2 were established from the same metastatic node and would appear to represent 2 clones of the tumour which differ in morphology, histochemical markers and DNA content. We present details of the morphology, DNA content and immunohistochemistry of these 4 lines and com-pare their growth patterns with those of SCLC and MCC lines which grow in suspension. © 1995 Wiley-Liss, Inc.