肿瘤坏死因子的调节性T细胞与肝癌发生的相关性

目的 探讨肿瘤坏死因子(tumor necrosis factor,TNF)的调节性T细胞与肝癌发生的相关性.方法 选取124例肝癌患者作为肝癌组,选择同期健康查体者124例作为对照组.检测2组血清TNF-α含量与肝功能,采用流式细胞仪检测外周血CD4+调节性T细胞比例并进行相关性分析.结果 肝癌组的血清TNF-α含量...     

Associations Between Three Common MicroRNA Polymorphisms and Hepatocellular Carcinoma Risk in Chinese

Aim: Associations between polymorphisms in miR-146aG>C, miR-196a2C>T and miR-499A>G and riskof HCC, and interaction with HBV infection in a Chinese population, were the target of the present research.Methods: The duplex polymerase-chain-reaction with confronting-two-pair primers (PCR-RFLP) was performedto determine the genotypes of the miR-146aG>C, miR-196a2C>T and miR-499A>G genotypes. Associations ofpolymorphisms with the risk of HCC were estimated by conditional logistic regression analysis. Results: Drinking,family history of cancer, HBsAg and HCV were risk factors for HCC. Multivariate regression analyses showedthat subjects carrying the miR-196a2 CC genotype had significantly increased risk of HCC, with an adjustedOR (95% CI) of 2.18 (1.23-3.80). In addition, cases carrying the miR-196a2 C allele had a 1.64-fold increase inthe risk for HCC (95%CI=1.03-2.49). The miR-196a2 CT and TT genotypes greatly significantly increased therisk of HCC in subjects with HBV infection, with adjusted ORs (95% CI) of 2.02 (1.12-3.68) and 2.69 (1.28-5.71),respectively. Conclusion: Our results demonstrate that miR-196a2 CC genotype and C allele have an importantrole in HCC risk in Chinese, especially in patients with HBV infection.     

抗原负载树突状细胞介导肝癌患者肿瘤杀伤细胞功能状态研究

目的:探讨肝癌细胞抗原负载树突状细胞激活肝癌患者外周血细胞毒性T细胞(CTL)、肿瘤浸润淋巴细胞(TI1)的抗肿瘤作用.方法:以细胞计数、间接荧光表型测定分析肝癌患者DC功能状态;MTT法测定肝癌细胞抗原负载DC介导肝癌患者外周血CTL、TIL对肝癌细胞的杀伤活性.结果:肝癌患者DC表达CD1a、CD80、CD83和HLA-DR等分子水平,DC介导的CTL、TIL对肝癌细胞的杀伤活性明显低于健康对照组(P＜0.05,P＜0.01).结论:肝癌患者存在DC功能缺陷,致使其介导的CTL、TIL对肝癌细胞的杀伤作用明显降低.     

肝细胞肝癌对机体细胞免疫的影响

目的　探讨原发性肝细胞肝癌(HCC )对机体细胞免疫机能的影响。方法　采用血常规检测,应用荧光标记抗体及三色激光流式细胞仪对5 1例肝细胞肝癌患者和17例对照样本术前、术后外周血的细胞免疫指标进行检测。检测项目包括:T辅助淋巴细胞(THL) ,细胞毒T淋巴细胞(CTL) ,NK细胞,单核细胞,并进行手术前后的对比分析。结果　原发性肝细胞肝癌(HCC)术后THL、CTL、NK和单核细胞百分比、单核细胞绝对值的平均值较术前分别增高0 .91% ,8.5 8% ,8.85 % ,3 1.45 % ,66.5 5 % ,手术前后有显著性差异(P <0 .0 5或P <0 .0 1)。良性对照组的术前与术后上述各项指标比较,均无显著性差异(P >0 .0 5 )。结论　肝细胞肝癌(HCC)对机体的细胞免疫产生明显抑制作用,在肿瘤切除之后,这种抑制能力随之减弱,细胞免疫得到恢复。而良性对照组对机体细胞免疫的抑制不显著。综合分析细胞免疫的变化情况,对判断HCC的预后及肿瘤复发有重要的意义。     

Relationship Between GSTT1 Gene Polymorphism and Hepatocellular Carcinoma in Patients from China

Objective: The results from studies on associations of the glutathione S-transferase T1 (GSTT1) genepolymorphism and hepatocellular carcinoma (HCC) risk in Chinese populations are still conflicting. This metaanalysiswas performed to evaluate the relationship in detail. Methods: Eligible reports were recruited into thismeta-analysis from the databases of PubMed, Embase, Cochrane Library and CBM-disc (China BiologicalMedicine Database). Results were expressed with odds ratios (OR) for dichotomous data, and 95% confidenceintervals (CI) were also calculated. Results: Eighteen investigations were identified for the analysis of associationbetween polymorphic deletion of GSTT1 and HCC, consisting of 2,693 patients with HCC and 4,696 controls.Null genotype of GSTT1 was associated with HCC susceptibility in Chinese (OR=1.53, 95%CI: 1.28-1.82;P﹤0.00001). Conclusion: The GSTT1 null genotype is associated with HCC susceptibility in Chinese.     

肝细胞癌与肝炎后肝硬化中多种肿瘤标志物的比较

目的探讨多种肿瘤标志物在肝细胞癌和肝炎后肝硬化中的表达情况。方法采用多种肿瘤标志物蛋白芯片检测系统测定85例肝细胞癌患者和92例肝炎后肝硬化患者血清中肿瘤标志物（CA199、CEA、CA242、FER、AFP、CA125、CA153）的水平,并比较各指标在两组间的差异情况。结果 7项肿瘤指标在肝细胞癌和肝炎后肝硬化中都有不同程度的阳性表达,且CA242和AFP在两组肝病间的比较有明显的统计学意义。结论多种肿瘤标志物的联合检测对肝细胞癌的诊断有较高的临床参考价值,适合于无明显症状的门诊患者的筛查和肿瘤高危人群的普查。     

肝癌热休克蛋白—抗原肽复合物的构建及诱导CTL反应的初步研究

目的：研究体外构建热休克蛋白-抗原肽复合的方法,观察其在体外的抗肿瘤作用。方法：用经43℃热处理1小时的人肝细胞悬液,经过裂解液裂解,60%-80%饱和硫酸铵沉淀,用SephadexG-100柱制备,取分子量为70KD组份,Westen blot进行性质鉴定。应用多肽解离液处理该组份,SephadexG-25柱过滤获得未结合多肽的蛋白分子,使其在体外与肝癌抗原肽SLIVHLNEV结合,构建成热休克蛋白-抗原肽复合物。应用此复合物树突状细胞,激活同源外周血T淋巴细胞产生肿瘤特异性杀伤T淋巴细胞（CTL）,应用MTT法检测其对T2细胞及肿瘤细胞的杀伤活性。结果：所得蛋白经电泳及Western blot进行蛋白分子量及性质鉴定为热休克蛋白70。SephadexG-25柱双分离法证实应用上述方法成功构建了肝癌热休克蛋白70-抗原肽复合物,用该复合物负荷树突状细胞在体外可以诱导出较强的肝癌抗原肽物特异性CTL,可以杀伤负荷有该肽的T2细胞及递呈该肽的肿瘤细胞系。结论：体外构建的热休克蛋白-抗原肽复合物可以增强抗原肽诱导CTL反应能力,热休克蛋白是良好的T细胞免疫佐剂,有可能在肿瘤疫苗治疗中发挥重要作用。     

Increased expression of immunosuppressive molecules on intratumoral and circulating regulatory T cells in non-small-cell lung cancer patients

The expression patterns of immunosuppressive molecules on regulatory T (Treg) cells have not been elucidated in non-small-cell lung cancer (NSCLC) patients. In this study, a total of 88 patients including 53 patients with NSCLC, 17 patients with lung non-malignant diseases, and 18 healthy volunteers were enrolled. Increased number of total CD4⁺CD25⁺FoxP3⁺ Treg cells and elevated expressions on the surface of several inhibitory molecules including CTLA-4, LAG-3 and PD-1 have been observed in the peripheral blood of NSCLC patients. We found that intratumoral Treg cells from NSCLC patients express the highest levels of co-inhibitory molecules compared to Treg cells isolated from tumor adjacent tissues or from peripheral blood of cancer patients, which is in consistent with the enhanced immunosuppressive function of these co-inhibitory molecules. Moreover, the number of Treg cells and their functional surface molecules increased during the progression of lung cancer. Elevated plasma levels of TGF-β and IL-10 in NSCLC patients were also observed in NSCLC patients compared to that in healthy volunteers. Our findings further support the role of Treg cells in the tumor microenvironments in NSCLC patients.     

Pathology and DNA Cytophotometry of Small Hepatocellular Carcinoma with a Nodule-in-nodule Appearance--Evidence for Stepwise Progression of Hepatocellular Carcinoma

Among 92 surgically resected human hepatocellular carcinomas(HCCs) 3cm in diameter, 23 tumors (25%) grossly showed a nodule-in-noduleappearance indicating stepwise progression of HCC. The centralnodules showed destructive growth of HCC, which was less differentiatedthan the surrounding area histologically, while the surroundingarea showed growth of very-welldifferentiated HCC with no, oronly minimal, destruction of the underlying liver structures.Previously, we proposed the term "early HCC" for HCCs withoutdefinite destructive growth, and the nodule-in-nodule lesionsdescribed here are considered to be in the transitional stagefrom early HCC to advanced HCC, and are therefore named earlyadvanced HCC (eAd HCC). DNA cytophotometry was performed in12 cases of eAd HCC, in which the early HCC component, advancedHCC component and non-tumorous liver tissue showing chronichepatitis or cirrhosis were analyzed separately. The early HCCcomponent showed a diploid pattern in eight of the 12 cases.The advanced HCC component showed an aneuploid peak in sevencases, and in three of these the peak was detected only in theadvanced HCC component and not in the early HCC component. Themean nuclear DNA content was significantly increased from nontumorousliver to the early HCC component and from the early HCC componentto the advanced HCC component. Polyploid cells containing morethan 4.8C DNA were exceptional in non-tumorous liver, but weredetectable in the early HCC component and increased in numberin the advanced HCC component. These findings suggest that DNAinstability may have an important role to play in the subclonalprogression of human HCC.     

Increased Frequency of Foxp3+ Regulatory T Cells in Mice with Hepatocellular Carcinoma

The CD4+CD25+ regulatory T cell (Treg) is a special kind of T cell subset. Studies have showed that Tregcells are involved in a number of physiological processes and pathologic conditions such as autoimmune diseases,transplantation tolerance and cancer. Tregs with unique capacity for immune inhibition can impair anti-tumourimmunity and help tumor cells to escape from immune surveillance. The aim of our study was to investigatewhether Tregs are involved in hepatocellular carcinoma (HCC). A BABL/C mouse with HCC in situ model wasestablished to evaluate the Treg existence in carcinoma tissues and the changes of Tregs in spleen using flowcytometry and immunohistochemistry methods. Granzyme B expression in carcinoma tissues was analyzedby immunohistochemistry to investigate the tumor local immune status.The proportion of CD4+CD25+/CD4+spleen lymphocytes of tumor bearing mice (18.8%±1.26%) was found to be significantly higher than that innormal mice (9.99%±1.90%) (P<0.01 ). Immunohistochemistry of spleen tissue also confirmed that there wasan increase in Treg in tumor-bearing mice, while in carcinomas it showed Treg cells to be present in tumorinfiltrating lymphocyte areas while Granzyme B was rarely observed. Anti-tumour immunity was suppressed,and this might be associated with the increase of Tregs. Our observations suggest that the CD4+CD25+Treg/CD4+ proportion in spleen lymphocytes can be a sensitive index to evaluate the change of Tregs in hepatocellularcarcinoma mice and the Treg may be a promising therapeutic target for cancer.     

Adoptive Transfer of Autologous Invariant Natural Killer T Cells as Immunotherapy for Advanced Hepatocellular Carcinoma: A Phase I Clinical Trial

Lessons Learned

• Administration of autologous invariant natural killer T (iNKT) cells was safe and well-tolerated in patients with hepatocellular carcinoma (Barcelona Clinic Liver Cancer stage B/C).
• Expanded iNKT cells produced T-helper 1–like responses with possible antitumor activity.
• No severe adverse events were observed in any of the enrolled patients, including one patient who received 10¹⁰ in vitro–expanded autologous iNKT cells as a single infusion.

     

A Case of Hepatocellular Carcinoma with Tumor Thrombi in the Epiploic Vein

A case of hepatocellular carcinoma (HCC) with tumor thrombiin the vein in the major omentum is reported. A 54-year-oldman underwent a second operation for recurrent HCC. Intraoperativeultrasonography revealed a tumor thrombus in the vein of theadhering omentum. Part of the antero-inferior area of the liveradjacent to the tumor was resected with the major omentum. Sinceit is characteristic of HCC often to form tumor thrombi in theveins, it is not surprising that the tumor budded into the epiploicvein. This is apparently the first report of tumor thrombi ofHCC in the epiploic vein.     

细胞浆内Connexin32蛋白增强肝癌细胞Li-7 的运动和侵袭能力*

目的：探讨肝癌细胞中间隙连接蛋白Connexin 32对肝癌细胞运动和侵袭能力的影响。方法：利用逆病毒感染的方法在肝癌细胞系Li-7 中建立Connexin 32蛋白表达可通过doxycycline 调控的Li-7 Tet-off Cx32稳定克隆,运用体外transwell migration and invasion assay方法验证Connexin 32表达与肝癌细胞运动和侵袭能力的相关性。结果：建立稳定整合了Connexin 32cDNA和调控序列Tet-off 的肝癌细胞Li-7 亚克隆,Western-blot结果显示外源性Connexin 32蛋白表达于细胞浆中,亚细胞定位分析证实Connexin 32蛋白定位于高尔基复合体内,此细胞浆内Connexin 32蛋白过表达显著增强肝癌细胞的体外运动和侵袭能力。结论：肝癌细胞浆内异位表达的Connexin 32蛋白发挥促进肿瘤进展的作用。     

TIL细胞与LAK细胞体外杀伤活力的比较

应用改良ＬＤＨ释放法，对ＴＩＬ细胞与ＬＡＫ细胞的前体及培养激活的细胞进行杀伤活力的检测，结果显示无论前体细胞或培养激活的细胞，对自体瘤细胞及Ｋ５６２细胞的杀伤活性均以ＴＩＬ细胞为高，表明ＴＩＬ细胞对肿瘤的特异识别及杀伤力均较ＬＡＫ细胞为强。     

肝细胞癌中NY-SAR-35 NY-TLU-57和NY-ESO-1基因的表达及意义

目的:检测三种癌-睾丸抗原NY-SAR-35、NY-TLU-57和NY-ESO-1基因mRNA在广西地区肝细胞癌(Hepatocellularcarcinoma,HCC)中的表达,探讨其作为HCC特异性免疫治疗靶抗原的可能性。方法:利用逆转录-聚合酶链反应(RT-PCR)方法,检测NY-SAR-35、NY-TLU-57和NY-ESO-1基因在63例HCC组织、56例HCC癌旁组织和4例正常肝组织中的表达,随机选择4例RT-PCR阳性产物直接进行DNA序列测定,并将其表达结果与临床病理指标进行统计学分析。结果:所检测的三种基因在4例正常肝组织中无表达;在63例HCC组织中,NY-SAR-35表达率为38.1%(24/63),NY-TLU-57为4.8%(3/63),NY-ESO-1为23.8%(15/63);在56例HCC癌旁组织中,NY-TLU-57和NY-ESO-1无表达,NY-SAR-35的表达率为26.8%(15/56)。基因表达与临床病理指标关系的分析显示,这三种癌-睾丸抗原的表达均与所分析的临床病理指标无关(P>0.05)。结论:癌-睾丸抗原NY-SAR-35、NY-TLU-57和NY-ESO-1基因可特异性的表达于HCC中,其中NY-SAR-35、NY-ESO-1具有一定的表达频率,提示它们有可能作为HCC特异性免疫治疗的靶抗原。     

抑癌基因甲基化谱分析在肝细胞癌诊断中的价值

目的 探讨beclin 1、RASSFA-1、p16、DAPK等抑癌基因启动子异常甲基化及其联合检测在肝癌早期诊断中的价值。方法 采用甲基化特异性PCR（methylation specific PCR,MSP）法,检测37例肝癌组织和相应的癌旁组织中beclin 1、RASSFA-1、p16和DAPK 4 种基因启动子区甲基化状态。通过与AFP的单一ROC曲线分析及其逐步Logistic回归的ROC曲线下面积,对单一及组合方式的敏感度、特异性、Youden指数和阳性似然比/阴性似然比进行比较分析。结果 肝癌组织中beclin 1、RASSFA-1、p16和DAPK 基因启动子区甲基化检出率分别为5.4%（2/37）、94.6%（35/37）、 73.0%（27/37 ）和 35.1% （13/37）,其中相应的癌旁组织RASSFA-1、p16和DAPK基因启动子区甲基化检出率与肝癌组织比较差异有统计学意义（P<0.05）,RASSFA-1基因启动子区甲基化和AFP肿瘤标志物联合检测可显著提高肝癌检测的敏感度（95.0%）和特异性（97.3%）,曲线下面积最大（0.903）,并获得高于单一基因甲基化的敏感度、Youden指数、阳性似然比/阴性似然比。结论 RASSFA-1基因启动子区甲基化和AFP肿瘤标志物联合检测,可能是早期筛查和诊断肝细胞癌的有效指标。     

Tumor Infiltrating Lymphocytes are Prognostic Factors and Can Be Markers of Sensitivity to Chemoradiotherapy in Head and Neck Squamous Cell Carcinoma

Background: Tumor-infiltrating lymphocytes (TILs) are assessed by the ratio of the area of lymphocytes infiltrating the stroma. TILs are important in breast cancer and malignant melanoma and are being established as a marker of prognosis and sensitivity to chemotherapy. This has resulted in various therapies being developed in fields such as breast cancer. However, the evaluation of TILs in head and neck squamous cell carcinoma (HNSCC) is not progressing, and the prognosis is still poor. Thus, investigating whether or not the evaluation of TILs is also effective in HNSCC and prognoses can be predicted with just biopsy samples alone is required. Methods: This study included 153 patients who were diagnosed with HNSCC between January 2010 and December 2019, underwent treatment, and could be followed up thereafter at our institution. Results: TILs, overall survival (OS), and progression-free survival (PFS) were evaluated in all patients, the chemoradiotherapy arm, and the surgery arm. The cut-off value for TILs was 50%. In all patients, OS was 69.8% and 40.2% (P = 0.01) and PFS was 58.4% and 31.6% (P = 0.003) in the high and low TIL groups, respectively. Multivariate analyses revealed that TILs independently predicted prognosis. In the chemoradiotherapy arm, OS was 70.8% and 31.6% (P = 0.012) and PFS was 63.4% and 20.3% (P = 0.001) in the high and low TIL groups, respectively. No significant differences were noted in the surgery arm. Conclusions: In HNSCC, TILs can be used as a prognosis predictor and chemoradiotherapy biomarker. Assessments can be performed just with hematoxylin–eosin staining and is very simple. This will greatly contribute to report personalized therapy progress. Further evaluations and, thus, prospective clinical multicenter trials are needed to use TILs in clinical practice for HNSCC.     

A Case of Fibrolamellar Hepatocellular Carcinoma

Fibrolamellar hepatocellular carcinoma is an uncommon histologic subtype of hepatocellular carcinoma, comprising 3.5% of all hepatocellular carcinomas in one large study (Brandt et al 1988). It usually occurs in young people, the average age being 18 years (Farhi et al 1983). It is not associated with cirrhosis and the serum alphafetoprotein is usually normal (Craig et al 1980). Fibrolamellar hepatocellular carcinoma has a more favourable prognosis than the usual hepatocellular carcinoma, with an average survival of 32 months from onset of symptoms, compared to 6 months for the usual type. The resectability rate and 5 year survival figures are also much better (Craig et al 1980). Hence distinguishing this histological subtype is important for surgical management and survival prognosis. This report describes a full radiological evaluation of such a case, covering the wide spectrum of relevant imaging modalities.     

肝细胞癌、鼻咽癌患者谷胱甘肽硫转移酶M1和T1基因型分布

目的 探讨高危区肝细胞癌和鼻咽癌患者谷胱甘肽硫转移酶M1 (GSTM1) 及T1 (GSTT1)基因多态性的分布。方法 应用PCR技术检测181例肝细胞癌、126例鼻咽癌患者和641例对照组人体GSTM1和GSTT1基因型。结果 GSTM1空白基因型(null)在肝癌组、鼻咽癌组与对照组频率分别为65.2%、61.9%和47.6%,病例组与对照组比较,差异有统计学意义（P＜0.01)。GSTT1空白基因型(null)在肝癌组、鼻咽癌组与对照组频率分别为57.5%、62.7%和43.1%,病例组与对照组比较,差异有统计学意义（P＜0.001)。结论 在肝细胞癌、鼻咽癌高发区解毒酶基因GSTM1和GSTT1呈多态性分布,二者的null基因型均增加患肝细胞癌、鼻咽癌的风险。     

Heterogeneity of Proliferative Activity in Nodule-in-Nodule Lesions of Small Hepatocellular Carcinoma

Surgically resected small hepatocellular carcinomas showing "nodule-in-nodule'formation were analyzed in terms of cell proliferative activity. The analysis was achieved by successful immunohistochemical demonstration of proliferating cell nuclear antigen in formalin-fixed paraffin-embedded tissue sections. Eight nodules (up to 3 cm in diameter) examined were either atypical adenomatous hyperplasia or hepatocellular carcinoma of low histologic grade, containing a discrete inner nodular area composed of obvious hepatocellular carcinoma of higher histologic grade. In all cases, the proliferating cell nuclear antigen labeling index of the latter area was much higher than that of the former, which in turn was slightly higher than that of the non-cancerous liver of the patient in 6 cases. The data presented here provide supporting evidence that the successive emergence and expansion of a more rapidly proliferating subclone within a nodule result in the stepwise progression of malignancy of human hepatocellular carcinoma.