Ex Vivo Interferon Gamma Production by Peripheral Immune Cells Predicts Survival in Lung Adenocarcinoma

BackgroundLung cancer is one of the most lethal malignancies, with a 5-year survival rate < 20% in patients with stage IV lung cancer. Impaired host immunity is associated with lung cancer pathogenesis, and interferon gamma (IFN-γ) plays an important role in antitumor immune surveillance. We evaluated the clinical significance of ex vivo production of IFN-γ in patients with lung adenocarcinoma.Patients and MethodsWe reviewed the medical records of 109 treatment-naive patients with lung adenocarcinoma who had undergone IFN-γ releasing assay. Differences in the IFN-γ level in nil and mitogen tubes were defined as ex vivo IFN-γ production. Correlation analysis was performed to evaluate the correlation between ex vivo IFN-γ production, cancer staging, and Eastern Cooperative Oncology Group performance status. The optimal cutoff values of low and high ex vivo IFN-γ production were estimated using receiver operator characteristic curve analysis. Cox proportional hazard analyses were used to evaluate the prognostic factors of 1-year overall patient survival.ResultsEx vivo IFN-γ production correlated with N stage, M stage, cancer staging, and Eastern Cooperative Oncology Group performance status. Low ex vivo IFN-γ production (ex vivo IFN-γ production ≤ 7.79 IU/mL) was independently associated with 1-year overall survival (odds ratio = 3.289; 95% confidence interval, 1.573-6.872; P = .002). Additionally, low ex vivo IFN-γ production was an independent predictor of 1-year overall survival in patients with stage IV cancer (odds ratio = 3.156; 95% confidence interval, 1.473-6.760; P = .003).ConclusionEx vivo IFN-γ production before treatment might be a useful biomarker for predicting prognosis in patients with lung adenocarcinoma.     

人结肠高分化腺癌细胞系THC—8908的建立及其生物学特性

本文报道将手术切除人结肠腺癌肿瘤标本,接种于裸鼠皮下,获可传代移植瘤,再体外培养,建立可连续培养的结肠高分化腺癌细胞系THC-8908,已传至160代。经光学和电镜检查该细胞系具有上皮性恶性细胞特征,放免检测证实具有分泌CEA能力,并观察了其体外生长曲线、分裂指数、软琼脂上集落形成率,染色体分析众数63,基本为亚三倍体,裸鼠皮下移植形成恶性种瘤,与手术标本病理形态相一致,是进行癌变机理研究和实验性基因治疗的良好模型。     

顺铂联合辐射对人盲肠未分化腺癌细胞系（HCe—8693）DNA含量的影响

本文应用显微分光光度术研究了顺铂联合辐射对人盲肠未分化腺癌细胞系(HCe-8693)DNA含量的影响.结果表明,药物组及药物联合辐射组随着实验后细胞培养时间的延长,细胞核 DNA含量逐渐下降,特别是药物联合辐射组,DNA含量呈不可逆减少.     

人白细胞干扰素对子宫内膜腺癌细胞系的作用研究

观察人白细胞干扰素对我们建立的子宫内膜腺癌细胞系的细胞生和细胞周期的影响。方法将不同浓度：２０００ｕ／ｍｌ、５００ｕ／ｍｌ、１００ｕ／ｍｌ的ＩＦＮ－α与ＪＥＣ共同培养７天,检测ＪＥＣ细胞生长曲线,细胞的凝集素受体表达,并用流式细胞仪分析细胞周期的变化,结果和结论ＩＦＮ－α对ＪＥＣ细胞的增殖有抑制作用,浓度越大,抑制作用明显,但不影响凝集素受体的表达;ＩＦＮ－α可使ＪＥＣ细胞Ｇ１／Ｇ０期增加、Ｓ期增     

白藜芦醇对结肠癌细胞生长的影响及作用机制探讨

背景与目的:探讨白藜芦醇(Rev)对结肠癌细胞系HT-29生长的影响,并结合细胞内环氧酶-2(COX-2)以及凋亡相关蛋白Bcl-2和Bax的表达情况探讨白藜芦醇的作用机制.材料与方法:分别以含有0、12.5、25、50、100 μmol/L白藜芦醇(Rev)的完全培养液培养HT-29细胞.用MTT法检测HT-29细胞生长情况,用western blot法测定细胞内COX-2的含量,用免疫细胞化学法检测细胞内凋亡相关蛋白Bel-2和Bax的表达情况. 结果:与对照组相比,Rev能够抑制HT-29细胞的生长,且呈剂量和时间依赖性.以不同浓度的Rev作用HT-29细胞48 h后发现,Rev对细胞内Bel-2的表达水平没有明显影响,但50和100 μmol/L的Rev能够明显抑制细胞内COX-2的表达,并且100 μmol/L的Rev能够明显上调细胞内Bax的表达.结论:Rev能够抑制结肠癌细胞HT-29的增殖,且具有剂量和时间依赖性,其作用与Rev能够抑制细胞内COX-2的表达以及上调Bax的表达有关.     

人子宫内膜癌细胞系HECCL-1的建立及其生物学特性的初步观察

目的 :建立人子宫内膜癌细胞系 ,并研究其生物学特性。方法 :将传代稳定后的人子宫内膜癌裸小鼠移植瘤标本用贴壁培养法建立细胞系 ,用光学显微镜、电子显微镜观察细胞系的形态 ,研究细胞系的生长及生物学特性。结果 :成功建立 1株人子宫内膜癌细胞系 ,已传代 6 0代 ,命名为HECCL 1。其生物学特征为 :形态学具有腺上皮癌细胞的特点 ,细胞生长旺盛 ,DNA为非整倍体 ,连续传代后仍保留人类肿瘤染色体的特点 ,异种动物移植阳性 ,雌、孕激素受体表达均阴性。结论 :HECCL 1细胞系的建立 ,为开展人类子宫内膜癌的基础及临床研究提供了理想的实验工具。     

洛铂体外诱导人结肠癌细胞株LOVO细胞凋亡及其作用机制

[目的]探讨洛铂体外诱导人结肠癌细胞株LOVO细胞凋亡及其作用机制。[方法]采用四甲基偶氮唑盐(MTT)法测定不同浓度(500μM、1000μM、2000μM)的洛铂作用于LOVO细胞24h的生长抑制率;原位末端标记技术法(TUNEL)检测LOVO细胞凋亡指数,计算凋亡阈值;流式细胞技术检测细胞周期变化;分光光度法检测半胱氨酸天冬氨酸蛋白酶(caspase)-3、8、9的表达。[结果]洛铂体外能抑制人结肠癌细胞株LOVO细胞的增殖,且呈剂量依赖性(P<0.05);洛铂能诱导LOVO细胞发生凋亡,凋亡阈值为1000μM;流式细胞技术分析显示洛铂致LOVO细胞周期明显阻滞在S期;分光光度法提示洛铂作用后LOVO细胞caspase-3、8、9表达明显高于阴性对照组(P<0.05)。[结论]洛铂能诱导体外培养的LOVO细胞凋亡,其作用机制可能与洛铂诱导LOVO细胞S期阻滞及促进caspase-8和caspase-9表达,进而促进cas-pase-3表达有关。     

PKC抑制剂对人肺腺癌细胞的放射增敏作用

 目的　探讨PKC抑制剂对肺癌放射敏感性的影响; 方法　采用MTT法检测了人肺腺癌细胞株A549在PKC抑制剂白屈莱红碱(CH)处 理前后,2Gy照射的细胞存活分数(SF2),并采用流式细胞术,对其凋亡率以及p53、Bcl -2蛋白表达进行了检测;结果　CH处理后A549凋亡 率增高,SF2下降。p53在X线照射后表达增强,而CH处理组p53表达 呈相对抑制状态。Bcl-2在处理前后变化温和。结论　PKC抑制剂对A 549细胞株的放射敏感性有增强作用,这种作用可能与凋亡率升高有关,而p53可能与 PKC抑制剂诱导的凋亡无关。     

In Silico Identification and Validation of Cuproptosis-Related LncRNA Signature as a Novel Prognostic Model and Immune Function Analysis in Colon Adenocarcinoma

Background: Colon adenocarcinoma (COAD) is the most common subtype of colon cancer, and cuproptosis is a recently newly defined form of cell death that plays an important role in the development of several malignant cancers. However, studies of cuproptosis-related lncRNAs (CRLs) involved in regulating colon adenocarcinoma are limited. The purpose of this study is to develop a new prognostic CRLs signature of colon adenocarcinoma and explore its underlying biological mechanism. Methods: In this study, we downloaded RNA-seq profiles, clinical data and tumor mutational burden (TMB) data from the TCGA database, identified cuproptosis-associated lncRNAs using univariate Cox, lasso regression analysis and multivariate Cox analysis, and constructed a prognostic model with risk score based on these lncRNAs. COAD patients were divided into high- and low-risk subgroups based on the risk score. Cox regression was also used to test whether they were independent prognostic factors. The accuracy of this prognostic model was further validated by receiver operating characteristic curve (ROC), C-index and Nomogram. In addition, the lncRNA/miRNA/mRNA competing endogenous RNA (ceRNA) network and protein–protein interaction (PPI) network were constructed based on the weighted gene co-expression network analysis (WGCNA). Results: We constructed a prognostic model based on 15 cuproptosis-associated lncRNAs. The validation results showed that the risk score of the model (HR = 1.003, 95% CI = 1.001–1.004; p < 0.001) could serve as an independent prognostic factor with accurate and credible predictive power. The risk score had the highest AUC (0.793) among various factors such as risk score, stage, gender and age, also indicating that the model we constructed to predict patient survival was better than other clinical characteristics. Meanwhile, the possible biological mechanisms of colon adenocarcinoma were explored based on the lncRNA/miRNA/mRNA ceRNA network and PPI network constructed by WGCNA. Conclusion: The prognostic model based on 15 cuproptosis-related lncRNAs has accurate and reliable predictive power to effectively predict clinical outcomes in colon adenocarcinoma patients.     

Effect of Buthionine Sulfoximine on the Sensitivity to Doxorubicin of Parent and MDR Tumor Cell Lines

Summary

We have studied the interaction of glutathione-depleting concentrations of buthionine sulfoximine (BSO) with the antiproliferative activity of doxorubicin (DXR) in three tumor lines, the mouse B16 melanoma, Friend erythroleukemia and the human K562 leukemia, both as DXR-sensitive and -resistant (with typical multidrug resistance) variants. BSO significantly enhanced the DXR effects in the wild-type Friend and K562 leukemias, and especially in the drug-resistant subline of Friend leukemia. BSO did not modify DXR accumulation and retention in the latter clone. Moreover, neither BSO nor verapamil used alone completely reversed the resistance to DXR of this cell line; their combination was more efficient and increased its drug sensitivity to a level closer to that of the parental counterpart. These results seem to indicate that the status of glutathione and of the enzymes related to it contributes to the resistance of Friend leukemia to DXR. An interesting additional finding was that BSO significantly synergizes with the antiproliferative effects of vincristine in the drug-sensitive variants of Friend and K562 leukemias.     

肺腺癌细胞A549中人survivin基因核心启动子位置与功能初探

[目的]探讨人survivin核心启动子的位置及在A549细胞中的转录激活功能。[方法]在生物信息学分析基础上,构建了4个分别由survivin 5′上游-1436~ 64、-612~ 64、-414~ 64和-203~ 64片段驱动的报告载体,分别瞬时转染A549及HK-2细胞,观察其在两种细胞的表达活性并比较在A549细胞中表达的活性差异。[结果]生物信息学提示survivin TSS位于ATG上游-64bp处,启动子近端富含若干潜在Sp1、p53和HIF-1α等结合位点。4种报告载体在A549细胞均有活性,且不同长度的启动子活性存在统计学差异。而在HK-2细胞中,4种载体未见显著性差异。[结论]人survivin启动子的活性具有肿瘤特异性,核心启动子可能位于TSS上游-203bp至 64bp处。suvivin核心启动子内可能存在2处HIF-1α的结合位点。     

EGFR Antisense Oligonucleotides Encapsulated withNanoparticles Decrease EGFR,MAPK1 and STAT5 Expressionin a Human Colon Cancer Cell Line

Epidermal growth factor receptor (EGFR) is over-expressed in several human cancers. This would suggestthat inhibition of EGFR is a reasonable approach for cancer treatment. In this study we investigated EGFRblocking and its effects on the mediated signaling such as MAPK and STATb in HT29 cells. For this aim weused FITC-labeled EGFR antisense oligonucleotides encapsulated with PAMAM nanoparticles to inhibit EGFRexpression. Cellular uptake of antisense was investigated by fluorescence microscopy and flow cytometry analysis.The effect of EGFR antisense on the expression of EGFR in HT29 cells was examined by real time PCR andWestern blots, which showed that antisense encapsulated with PAMAM decreased the level of EGFR mRNAand protein. In addition, real time PCR results confirmed that EGFR inhibition had an effective role in thereduction of EGFR dependent downstream genes. In conclusion, EGFR antisense encapsulated with PAMAMnanoparticles down regulated EGFR and EGFR-mediated genes.     

人肺腺癌诺维本耐药细胞株的耐药机制

 研究人肺腺癌诺维本耐药细胞系Anip973/NVB的耐药机制。 方法 免疫组织化学法检测人肺腺癌诺维本耐药细胞系的P-糖蛋白（P-gp）、多药耐药相关蛋白(MRP)的表达,逆转录 聚合酶链反应（RT-PCR）法检测MDR1基因以及MRP3基因的转录。 结果 Anip973/NVB中MRP呈强阳性表达,而Anip973未见表达。Anip973/NVB细胞中MRP3的mRNA转录增加,未检测到MDR1的mRNA产物（Ｐ<0.05）。 结论 MRP3在人肺腺癌诺维本耐药细胞中高表达,说明诺维本的耐药机制可能与MRP3的表达相关。     

常规化疗药物连续刺激对人肺腺癌细胞株表达erbB-2，p53和MDR1的影响

目的:分析常规化疗药物连续刺激对肿瘤细胞株表达erbB-2,p53和MDR1的影响,为联合应用生物治疗提供实验基础.方法:人肺癌细胞株经ADM,VP-16,DDP单药及联合用药连续刺激,每种药物分2个剂量梯度.应用流式细胞术分别检测erbB-2,p53和MDR1的阳性细胞数及平均荧光强度,以此推算出不同药物在不同浓度下连续2～3次作用后细胞株以上各蛋白表达的细胞数、平均表达量、总表达量,同时设对照组.结果:随着培养时间的延长各项检测指标其阳性细胞的百分数、平均荧光强度及总荧光强度均呈下降趋势.高剂量各组erbB-2和MDR1以上各项指标基本上均随刺激次数的增加呈下降趋势,而低剂量各组的检测指标则有升有降;p53的表达无一定规律,但第三次刺激后药物组的表达均高于对照组.结论:化疗药物连续刺激后,特别是小剂量化疗后,erbB-2,p53和MDR1的表达可不同程度增高,提示临床上针对这些分子对病人实 施生物治疗时应考虑化疗药物的影响.     

MDR1基因下调逆转人白血病阿霉素耐药细胞株K562/ADM的耐药性

目的：探讨RNA干扰（RNAi）人MDR1基因对人白血病阿霉素耐药细胞株K562/ADM耐药性的影响。方法：应用针对人MDR1基因的RNAi质粒pENTRTM/U6-MDR1转染人白血病阿霉素耐药细胞株K562/ADM和亲本细胞株K562,48 h后实时荧光定量PCR检测MDR1 mRNA表达,流式细胞术检测P-gp蛋白表达和P-gp功能,MTT法检测细胞对ADM的耐药性。结果：与未转染细胞相比,K562/ADM耐药细胞pENTRTM/U6-MDR1组的MDR1 mRNA和P-gp蛋白表达和功能均显著下降（ P <0.05）,对阿霉素的耐药性显著降低（ P <0.05）。结论：MDR1基因下调可逆转人白血病阿霉素耐药细胞株对阿霉素的耐药性。     

塞来昔布对人结肠癌细胞VEGF-C表达的影响

[目的]探讨塞来昔布对体外培养的结肠癌细胞血管生长因子C（VEGF-C）表达的影响。[方法]以人结肠癌细胞HT29为研究对象,以不同浓度的塞来昔布（0、25、50、100μmol/L）进行处理。采用RT-PCR等方法检测四组细胞中VEGF-C的表达情况。[结果]塞来昔布可抑制人结肠癌HT29细胞VEGF-C的表达,抑制作用与塞来昔布的浓度相关。[结论]塞来昔布可能通过对肿瘤细胞中VEGF-C基因表达的抑制发挥其抗肿瘤作用。     

人结肠高分化腺癌裸小鼠移植瘤模型的建立及其生物学特性

将人结肠高分化腺癌手术标本，接种于裸小鼠体内建成移植瘤ＴＨＣＮ－８９０１，已传１４代，历时２年多，生长稳定，可移植性成功率高。组织学及超微结构形态观察，均保持了原病人肿瘤的结构，并且有分泌功能，染色体分析证实为人染色体，众数在６３～９４之间，该瘤株具有稳定产生ＣＥＡ的生物学特性，用＾１１１Ｉｎ－ＣＥＡＭｃＡｂ能使裸小鼠移植瘤清晰显像。此瘤株的建立为结肠癌癌变基础及实验性基因治疗研究，提供了较为理想