Highly Efficient Labeling of Human Lung Cancer Cells Using Cationic Poly-L-lysine-Assisted Magnetic Iron Oxide Nanoparticles

Cell labeling with magnetic iron oxide nanoparticles(IONPs)is increasingly a routine approach in the cellbased cancer treatment.However,cell labeling with magnetic IONPs and their leading effects on the biological properties of human lung carcinoma cells remain scarcely reported.Therefore,in the present study the magnetic c-Fe2O3nanoparticles(MNPs)were firstly synthesized and surface-modified with cationic poly-L-lysine(PLL)to construct the PLL-MNPs,which were then used to magnetically label human A549 lung cancer cells.Cell viability and proliferation were evaluated with propidium iodide/fluorescein diacetate double staining and standard 3-(4,5-dimethylthiazol-2-diphenyl-tetrazolium)bromide assay,and the cytoskeleton was immunocytochemically stained.The cell cycle of the PLL-MNPlabeled A549 lung cancer cells was analyzed using flow cytometry.Apoptotic cells were fluorescently analyzed with nuclear-specific staining after the PLL-MNP labeling.The results showed that the constructed PLL-MNPs efficiently magnetically labeled A549 lung cancer cells and that,at low concentrations,labeling did not affect cellular viability,proliferation capability,cell cycle,and apoptosis.Furthermore,the cytoskeleton in the treated cells was detected intact in comparison with the untreated counterparts.However,the results also showed that at high concentration(400 lg m L-1),the PLL-MNPs would slightly impair cell viability,proliferation,cell cycle,and apoptosis and disrupt the cytoskeleton in the treated A549 lung cancer cells.Therefore,the present results indicated that the PLL-MNPs at adequate concentrations can be efficiently used for labeling A549 lung cancer cells and could be considered as a feasible approach for magnetic targeted anti-cancer drug/gene delivery,targeted diagnosis,and therapy in lung cancer treatment.

Continuous intravenous infusion of recombinant human endostatin using infusion pump plus chemotherapy in non-small cell lung cancer

BACKGROUND Lung cancer is one of the deadliest cancers in the world with the highest incidence and mortality rate among all cancers.Non-small cell lung cancer(NSCLC)accounts for approximately 80%of primary lung cancer.However,efficacy and safety of the current regimens for NSCLC is unsatisfactory.Therefore,there has been an increasing urgency for development of potential therapeutic therapies for NSCLC.AIM To investigate the therapeutic outcomes and safety of continuous intravenous infusion of recombinant human endostatin(Rh-endostain)using an infusion pump in retreated advanced NSCLC.METHODS Patients with retreated advanced NSCLC who were admitted to Zhejiang Provincial People's Hospital from October 2017 to April 2019 were recruited.These patients received continuous intravenous infusion of Rh-endostain using an infusion pump.Objective response rate(ORR),clinical benefit rate(CBR),median progression-free survival(mPFS),and incidences of adverse events(AEs)were analyzed after treatment.RESULTS A total of 45 patients with retreated advanced NSCLC were included,and all of them were evaluated.In these patients,ORR was 22.2%,CBR was 84.4%,and mPFS was 5.3 mo.The following AEs were observed,decreased hemoglobin(34 cases,75.6%),nausea/vomiting(32 cases,71.1%),elevated transaminase(24 cases,53.3%),leukopenia(16 cases,35.6%),thrombocytopenia(14 cases,31.1%),and constipation(1 case,3.4%).None of the patients had leukopenia,nausea/vomiting,and constipation of grade III and above.CONCLUSION The patients showed improved adherence to 5-d continuous intravenous infusion of Rh-endostain using an infusion pump.Favorable efficacy and safety of this treatment regimen were achieved in retreated advanced NSCLC.

Alterations and Its Mechanisms of Wnt Signal Pathway in Human High-matastatatic Large Cell Lung Cancer Cell Line L9981 by Transfecting with Nm23-H1 Gene

Backgroud and Objective Tumor metastasis is not only the malignant marker and characteristics of lung cancer, but also the main cause of failure to cure and lose their life of the

ANTI-HUMAN LUNG GIANT CELL CANCER (PG) EFFECT OF HUMAN LAK CELLS IN VITRO AND IN NUDE MICE

Human LAK cells were prepared by culturing normal human peripheral blood mononuclear cells (PBMC) with or without rIL-2 and assayed for T cell surface markers as well as anti-tumor activity against PC in vitro and in nude mice. Although the percentages of T3, T4, and T8 positive cells in rIL-2-activated cells did not differ significantly from those of control cells in vitro, the former showed stronger cytotoxicity than control cells to PG tumor cells in vitro. In vivo, LAK cells completely inhibited the growth of PG tumor in nude mice, whereas PBMC control cells were to be of no effect. The anti-tumor effect of human LAK cells in nude mice may offer a useful model to study the role of human LAK cells against human tumor in vivo.

Effects of recombinant human endostatin on therapeutic effect, angiogenesis, tumor cell proliferation and migration in patients with non-small cell lung cancer

Objective:To investigate the effects of recombinant human endostatin on therapeutic effect, angiogenesis, tumor cell proliferation and migration in patients with non-small cell lung cancer.Methods: A total of 100 patients with non-small cell lung cancer treated in our hospital from September 2015 to March 2018 were selected as research subjects.They were randomly divided into observation group and control group, 50 cases in each group.The control group was treated with gemcitabine plus cisplatin, while the observation group was combined with Endostar treatment on the basis of the control group. Observed and compared the expression of the therapeutic effect[including cytokeratin 19 fragment (CYFRA21-1), carcinoembryonic antigen (CEA) and carbohydrate antigen (CA50)], angiogenesis[including vascular endothelial growth factor (VEGF) and hypoxia inducible factor (HIF-1 alpha)], tumor cell proliferation[including insulin-like growth factor (IGF-1) and insulin-like growth factor receptor -7 (IGFBP-7)] and migration[including high mobility group protein AT-hook2 (HMGA2) and high mobility group protein B1 (HMGB1)]in two groups.Results: The two groups showed significant changes in the therapeutic effect, angiogenesis, proliferation and migration of tumor cells. Before treatment, there was no significant difference in all levels between the two groups. After treatment, the levels of CYFRA21-1, CEA, CA50, VEGF, HIF-1a, IGF-1, HMGA2 and HMGB1 in the two groups were significantly lower than those before treatment, while the levels of IGFBP-7 were significantly higher than those before treatment. After treatment, the levels of CYFRA21-1, CEA, CA50, VEGF, HIF-1a, IGF-1, HMGA2 and HMGB1 in the observation group were significantly lower than those in the control group, while the levels of IGFBP-7 were significantly higher than those in the control group. Conclusions: Recombinant human endostatin can enhance the therapeutic effect of non-small cell lung cancer patients, reduce angiogenesis, inhibit tumor cell proliferation and migration.

The Difference of Gene Expression Prof ile in Human Large Cell Lung Cancer Cell Lines with Different Metastatic Potential

Background and Objective Lung cancer is the most lethal malignangy that threatens human heath and lives nowadays in the world, and meanwhile is also the one with worst

The Difference of the Copy Number Variation and Loss of Heterozygosity of Human Lung Large Cell Cancer Cell Line with Different Metastatic Potential

Background and Objective It has been proven that copy number gain/or loss (copy number variation CNV) in uences gene expression and result in phenotypic variation by

The Effects of Intracellular Glutathione Content on the Sensitivity of Methylseleninic Acid to Human High-metastatic Large Cell Lung Cancer Cell Line L9981

Background and Objective Lung cancer has the fastest increasing rate of morbidity and mortality all over the world and appears to be one of the most dangerous malignant tumors

Effects of Methylseleninic Acid on the Proliferation and Cell Cycle in Human High Metastatic Large Cell Lung Cancer Cell Line L9981 and Its Molecular Mechanism

Background and Objective Lung cancer is the rst killer of human being in the whole world. Recently, although many treatment strategies have been developed, the anti-cancer effects

Down-regulation and It's Molecular Biology of Nm-23H1 gene Transfection on Ras-to-MAPK Signal Transduction Pathway in Human High-metastatic Large Cell Lung Cancer Cell Line L9981

Backgroud and Objective Tumor metastasis is not only the malignant marker and characteristics of lung cancer, but also the key cause of failure to cure and lose their life of the patients

Influence of Nm23-H1 Gene Site Mutagenesis on Invasive And Metastatic Phenotype in Human High-Metastatic Large Cell Lung Cancer Cell Line L9981

Background and Objective Invasion and metastasis is not only the malignant phenotypes of lung cancer but also the main cause of death. To study and elucidate the molecular mechanism

蛇葡萄素对人肺癌GLC-82裸鼠移植瘤的抑制作用

目的 :为了进一步研究蛇葡萄素体内抗肿瘤作用 ,我们进行了人肺癌GLC 82裸小鼠移植瘤的抑瘤实验研究。方法 :人肺癌GLC 82细胞接种于裸鼠腋窝建立移植模型。荷瘤BALB/C裸鼠随机分成 6组 ,蛇葡萄素以 3个剂量腹腔给药。观察肿瘤体积、相对肿瘤体积、瘤重、相对肿瘤增殖率以及肿瘤生长曲线 ,以此评价蛇葡萄素的抗瘤作用。结果 :2 5 0mg/kg蛇葡萄素对裸鼠移植瘤的抑制率 ,二次实验结果分别为 35 5 % (P <0 0 1)和 37 1% (P<0 0 1) ,相对肿瘤增殖率则分别为 5 5 2 4 % (P <0 0 1)和 5 7 71% (P <0 0 5 )。结论 :蛇葡萄素对人肺癌GLC 82裸鼠移植瘤具有显著的抑制作用。

Brefeldin A通过内质网应激途径协同增强顺铂抗肺癌细胞GLC-82作用

已有研究证明,内质网应激(endoplasmic reticulum stress,ERS)启动的未折叠蛋白质反应(unfolded protein response,UPR)信号途径有助细胞存活;然而,长时间剧烈ERS可诱导细胞凋亡,因此,ERS是肿瘤治疗的新靶点。抗癌药物具有严重的毒副作用,而药物的协同作用是减少抗癌药物使用剂量、减少毒副作用的重要手段之一。本研究证明布雷菲德菌素A(brefeldin A,BFA)与顺铂(cis-dichlorodiamine platinum,CDDP)的协同抗肺癌细胞的作用。MTT试验显示,单独应用BFA和顺铂时,它们抑制肺癌细胞GLC-82生长的半数有效浓度(IC50)分别是100 ng/m L和4μg/m L;而采用半量的BFA和CDDP联合处理GLC-82细胞24 h后,其抑制生长作用进一步加强;等效线图解法及联合作用指数分析进一步证明二者具有协同作用。两药的这种协同作用进一步被形态学检查证明——与单独用药比较,除了细胞皱缩,出现了更多的染色质固缩,细胞质及细胞核裂解碎片,乃至形成凋亡小体,提示细胞凋亡的发生。实时定量PCR及蛋白质印迹证明,与单独用药比较,联合用药导致内质网应激标志分子GRP78、CHOP表达水平明显增加,提示有内质网应激通路激活。上述结果证明,BFA与CDDP联合用药可增强对肺癌细胞GLC-82的生长抑制作用,其协同机制可能与内质网应激通路的激活有关。我们的结果支持"ERS是肿瘤治疗新靶点"学说,对肺癌的新化疗方案有一定的指导意义。

当归挥发油对人肺腺癌GLC-82细胞增殖及凋亡的影响

目的:探讨当归挥发油对人肺腺癌GLC-82细胞增殖及凋亡的作用。方法:MTT比色法检测当归挥发油对GLC-82细胞的细胞毒作用;流式细胞术检测GLC-82细胞凋亡;吖啶橙/溴化乙锭(AO/BE)双荧光染色法观察GLC-82细胞凋亡。结果:当归挥发油在浓度6.25~200μg/m L范围内对GLC-82细胞的增殖具有浓度依赖性抑制作用,对GLC-82细胞的IC_(50)为113.03μg/m L。流式细胞术检测结果表明,当归挥发油可诱导GLC-82细胞凋亡、坏死,且细胞凋亡、坏死率随药物浓度增高而升高。AO/BE双荧光染色法结果显示,用药组随着药物浓度的升高,细胞结构固缩加重,凋亡、坏死细胞增多。结论:当归挥发油具有诱导GLC-82细胞凋亡的作用。

蛇葡萄素钠联合卡铂对人肺腺癌GLC-82细胞增殖及凋亡的影响

目的探讨蛇葡萄素钠(AMP-Na)单用及其与卡铂(CBP)合用对人肺腺癌GLC-82细胞增殖的抑制作用及可能的机制。方法四甲基偶氮唑盐(MTT)比色法测定两药单用及合用对GLC-82细胞的体外杀伤活性;透射电子显微镜(TEM)观察GLC-82细胞超微结构的变化;流式细胞仪(FCM)检测GLC-82细胞凋亡和caspase-3的表达。结果对于GLC-82细胞,AMP-Na与CBP合用,CBP的IC50由(17.10±4.78)mg·L-1降低到<3.12 mg·L-1(P<0.01),表明Amp-Na对卡铂抑制GLC-82细胞增殖的作用具有协同效应(CDI<1)。TEM及FCM检测结果表明,单用AMP-Na及与卡铂合用均可诱导GLC-82细胞凋亡、坏死,两药合用后坏死率由(2.56±0.41)%升高到(71.83±5.43)%(P<0.01)。AMP-Na单用及与卡铂合用作用于GLC-82细胞48 h后,caspase-3的表达均明显增高。结论AMP-Na和CBP具有协同诱导GLC-82细胞凋亡的作用,机制可能与激活细胞内caspase-3介导的信号转导通路相关。

当归挥发油对人肺腺癌GLC-82细胞增殖及细胞周期的影响

目的:探讨当归挥发油对人肺腺癌GLC-82细胞增殖及细胞周期的作用。方法:采用噻唑蓝(MTT)比色法检测当归挥发油对GLC-82细胞存活率的影响;倒置显微镜下观察细胞生长形态;流式细胞仪检测当归挥发油对GLC-82细胞周期的影响。结果:在6. 25～200μg/m L浓度范围内当归挥发油对GLC-82细胞增殖具有浓度依赖性抑制作用,当归挥发油对GLC-82细胞的I C50为113. 03μg/mL;倒置显微镜下观察发现阴性组细胞具有GLC-82细胞典型的形态特征,多边形,成片生长,随着当归挥发油浓度的升高,细胞数量减少,散在生长,形态发生变化,逐渐皱缩;经当归挥发油处理后GLC-82细胞表现出明显的G2期周期阻滞。结论:当归挥发油可抑制GLC-82细胞生长,引起细胞周期G2期的阻滞。

血管性血友病因子及整合素β_3与人肺癌细胞GLC-82黏附的初步研究

目的观察血管性血友病因子(vWF)及整合素β(3integrin β3)在人肺癌细胞株GLC-82的表达,人肺癌细胞株GLC-82与vWF黏附的关系,及vWF与integrinβ3之间的相互关系。方法应用免疫组织化学技术观察vWF及integrin β3在人肺癌细胞株GLC-82的表达;应用黏附实验、抗体抑制实验及MTT法,观察人肺癌细胞株GLC-82与vWF的黏附,及vWF与integrinβ3的相互关系。结果免疫组织化学结果显示GLC-82细胞均表达vWF及整合素β3;GLC-82细胞可以与vWF黏附,Anti-integrinβ3抑制后这种黏附降低,降低效果与Anti-vWF抑制黏附相同。结论GLC-82细胞表达vWF及integrinβ3,GLC-82细胞可以与vWF黏附,从而促进肿瘤细胞转移,vWF可能通过与integrinβ3结合发挥作用。

Inhibition Mechanism of Novel Pyrazolo[1,5-a]pyrazin-4(5H)-one Derivatives Against Proliferation of A549 and H322 Cancer Cells

Objective To explore the inhibition mechanism and safety of pyrazolo[1,5-a]pyrazin-4(5H)-one derivatives against proliferation of human lung cancer A549 cells, H322 cells, and human umbilical vein endothelial cell(HUVEC). Methods Cells were treated with 40 μmol/L of the ppo3 a, ppo3 b, ppo3 i, and 0.1% DMSO(control) for 48 hours, respectively. Apoptosis was determined by Hoechst 33258 staining assay in H322 and A549 cells. Cell cycle distribution was determined by flow cytometry analysis in A549 cell. LC3-II, p53, and heat shock protein(HSP) 70 protein levels were detected by Western blotting in A549 cells treated with ppo3 b for 48 hours. The morphology and viability of HUVEC were observed by inverted microscope and sulforhodamine B(SRB) assay. Results Ppo3 a, ppo3 b, and ppo3 i significantly induced apoptosis in H322 and A549 cells. A strong G1-phase arrest was concomitant with the growth inhibitory effect on A549 cells. Ppo3 b effectively elevated the p53 protein level, but significantly reduced the HSP70 protein level. There were no significantly inhibitory effect on the morphology and viability of HUVEC when treated with ppo3 a, ppo3 b, and ppo3 i. Conclusions ppo3 a, ppo3 b, and ppo3 i could inhibit H322 proliferation through apoptosis and inhibit A549 through apoptosis and G1-phase arrest. The protein p53 and HSP70 might involve in the inhibition effects. These derivatives might be a clue to find effective and safe drug for lung cancers.

Value of serum NGAL and HE4 content change for diagnosing lung cancer and evaluating disease progression

Objective: To study the value of serum NGAL and HE4 content change for diagnosing non-small cell lung cancer (NSCLC) and evaluating disease progression. Methods: Patients who were diagnosed with NSCLC and underwent surgical resection in the First Affiliated Hospital of Chengdu Medical College between February 2015 and March 2017 were selected as the NSCLC group, and healthy volunteers who received physical examination over the same period were selected as control group. The NGAL and HE4 contents in serum of NSCLC group and control group as well as the expression of proliferation, apoptosis and invasion genes in NSCLC lesion of NSCLC group were detected. Results: Serum NGAL and HE4 levels of NSCLC group were significantly higher than those of control group, serum NGAL and HE4 levels of patients with TNM II and TNM III NSCLC were significantly higher than those of patients with TNM I NSCLC, and serum NGAL and HE4 levels of patients with TNM III NSCLC were significantly higher than those of patients with TNM II NSCLC;c-myc, Survivin, CatL, Vimentin, Slug and Snail mRNA expression in NSCLC lesion were significantly higher than those in adjacent lesion and positively correlated with serum NGAL and HE4 levels while PTEN, LATS1 and E-cadherin mRNA expression were significantly lower than those in adjacent lesion and negatively correlated with serum NGAL and HE4 levels. Conclusion: Abnormally elevated NGAL and HE4 in the serum of NSCLC patients can evaluate the proliferation and invasion of cancer cells to a certain extent.

重组人血管内皮抑制素联合奥希替尼治疗晚期非小细胞肺癌的疗效及预后分析

目的:分析重组人血管内皮抑制素联合奥希替尼对晚期非小细胞肺癌(non-small cell lung cancer, NSCLC)患者疗效及预后的影响。方法:选择2020年06月至2022年02月医院就诊的晚期NSCLC患者92例,采用随机数字表法将其分成对照组与研究组,各46例。对照组接受奥希替尼治疗,研究组在对照组的基础上接受重组人血管内皮抑制素治疗。21 d为1个周期,2组均治疗3个周期评估效果。对比2组临床疗效、健康状况、生活质量、肿瘤标志物、肿瘤相关蛋白因子及不良反应,随访1年,记录2组无进展生存期(PFS)。结果:研究组客观缓解率(50.00%)高于对照组(26.09%)(P<0.05),研究组疾病控制率(76.09%)高于对照组(54.35%)(P<0.05)。治疗3个周期后,2组卡氏功能状态(KPS)评分、癌症治疗功能性量表(FACT-L)评分均升高(P<0.05),且研究组更高(P<0.05)。治疗3个周期后,2组血清细胞角蛋白19片段(CYFRA21-1)、鳞状细胞癌抗原(SCC)、糖类抗原50(CA50)水平均降低(P<0.05),且研究组更低(P<0.05)。治疗3个周期后,2组磷酸酶张力蛋白同源物(PTEN)相对表达量均升高(P<0.05),且研究组更高(P<0.05);治疗3个周期后,2组血黏蛋白(MUC1)相对表达量均降低(P<0.05),且研究组更低(P<0.05)。2组Ⅰ-Ⅳ级消化道反应、血小板下降、肝肾功能损伤、中性粒细胞减少不良反应总发生率比较差异无统计学意义(P>0.05)。随访1年,2组均失访1例,随访率为97.83%,研究组中位PFS为8.97(95%CI:6.13~11.35)个月,对照组中位PFS为6.53(95%CI:3.85~9.61)个月,研究组PFS曲线优于对照组(P<0.05)。结论:重组人血管内皮抑制素联合奥希替尼治疗晚期NSCLC患者疗效确切,可降低肿瘤标志物水平,改善患者生活质量,调节肿瘤相关蛋白因子表达,安全可靠,且可延长患者PFS。