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

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。

miR-519d在非小细胞肺癌脑转移中的作用及机制

目的探究miR-519d在非小细胞肺癌(NSCLC)脑转移(BM)中的作用及其机制。方法采用RT-PCR检测miR-519d在伴有BM的NSCLC患者和无BM的患者癌旁正常肺组织、NSCLC肺组织、NSCLC脑组织,人NSCLC细胞系(SK-MES-1、NCI-H226、NCIH1299和NCL-H820)、正常人支气管上皮细胞(HBECs)、人脑微血管内皮细胞(HBMECs)及正常人星形胶质细胞(NHA)中的表达。将过表达miR-519d慢病毒(OE-miR-519d-LV)或空载体慢病毒(NC-LV)转染至NSCLC细胞系NCI-H1299中,作为OE-miR-519d-LV组和NC-LV组;联合转染OE-miR-519d-LV与pcDNA3.1-HER3的细胞作为OE-miR-519d-LV+pcDNA3.1-HER3组。将OE-miR-519d-LV组和NC-LV组细胞经心脏注射至裸鼠体内,以构建伴BM的NSCLC动物模型,并按注射细胞的不同将小鼠分为Ctrl组与miR-519d组。使用生物发光成像系统观察NCI-H1299细胞在小鼠大脑中的转移情况。CCK-8法检测OE-miR-519d-LV组和NC-LV组中NCI-H1299细胞的增殖情况。采用HBMECs和NHA包被的Transwell小室,以建立体外血脑屏障(BBB)细胞模型,并检测OE-miR-519d-LV组和NC-LV组NCI-H1299细胞跨内皮细胞的迁移能力。Western blot检测各组细胞中上皮间质转化(EMT)相关蛋白MMP-2、MMP-9、E-cadherin、N-cadherin和Vimentin的表达。生物信息学技术分析miR-519d与HER3的潜在结合位点,双荧光素酶基因报告实验验证两者的靶向调控关系。结果与癌旁正常肺组织相比,miR-519d在NSCLC肺组织和NSCLC脑组织中的表达降低(P<0.05);与NSCLC肺组织相比,NSCLC脑组织中miR-519d表达降低(P<0.05)。与HBECs相比,miR-519d在NSCLC细胞系中的表达降低(P<0.05),且miR-519d在NCI-H1299、NCI-H226和NCL-H820细胞中的表达水平低于SK-MES-1细胞(P<0.05)。与Ctrl组裸鼠相比,miR-519d组裸鼠的BM发生率更低,生存时间更长,差异有统计学意义(P<0.05)。与NC-LV组细胞相比,OE-miR-519d-LV组NCI-H1299细胞增殖活力、跨内皮细胞迁移能力及MMP-2、MMP-9、N-cadherin和Vimentin蛋白表达降低(P<0.05),而E-cadherin蛋白表达升高(P<0.05)。miR-519d可靶向调控HER3蛋白表达。与OE-miR-519d-LV组相比,OE-miR-519d-LV+pcDNA3.1-HRE3组细胞的迁移能力升高(P<0.05),MMP-2、MMP-9、N-cadherin和Vimentin蛋白表升高(P<0.05),而E-cadherin表达降低(P<0.05)。结论miR-519d在伴有BM的NSCLC中表达水平降低,上调其表达可通过靶向调控HER3来抑制肿瘤的BM。

重组人血管内皮抑制素注射液联合GP化疗方案治疗晚期NSCLC的可行性

目的观察重组人血管内皮抑制素注射液联合GP化疗方案治疗晚期非小细胞肺癌(NSCLC)的可行性。方法回顾性选取2020年5月—2022年12月皖北煤电集团总医院肿瘤内科收治的晚期NSCLC患者80例,将采用GP化疗方案治疗者纳入GP化疗组(n=40),重组人血管内皮抑制素注射液联合GP化疗方案治疗者纳入联合用药组(n=40),2组均以3周为1个治疗周期,共治疗2个周期。比较2组患者近期疗效,用药前后实验室指标、生活质量评分,不良反应及1年存活率。结果联合用药组客观缓解率为85.00%,高于GP化疗组的52.50%(χ^(2)=9.833,P=0.002)。用药2个周期后,2组血清癌胚抗原、糖类抗原125水平低于用药前,凝血酶时间、活化部分凝血活酶时间短于用药前,凝血酶原时间长于用药前,血浆纤维蛋白原、D-二聚体水平高于用药前,且联合用药组各指标优于对照组(P<0.05或P<0.01);2组生理功能、躯体功能、精神状态、社会关系评分高于用药前,且联合用药组高于GP化疗组(P<0.01)。联合用药组与GP化疗组不良反应总发生率分别为22.50%、15.00%,组间比较差异无统计学意义(χ^(2)=0.738,P=0.390)。联合用药组1年存活率为65.00%,高于GP化疗组的42.50%(χ^(2)=4.073,P=0.044)。结论晚期NSCLC治疗中采用重组人血管内皮抑制素注射液联合化疗药物的可行性较高,可有效降低肿瘤标志物水平,延缓病情进展,提高患者生活质量及1年存活率,且具有一定安全性。

持续静脉泵注重组人血管内皮抑制素联合化疗一线治疗晚期非小细胞肺癌的随机、对照临床试验

目的探讨持续静脉泵注重组人血管内皮抑制素联合化疗一线治疗晚期非小细胞肺癌(Non-Small Cell Lung Cancer,NSCLC)的随机、对照临床试验。方法选取2019年3月—2022年9月期间雅安市人民医院接受治疗的150例晚期NSCLC患者为研究对象,根据随机数表法划分成两组,每组75例,化疗组给予常规一线化疗方案治疗,联合组在化疗组基础上加以持续静脉泵注重组人血管内皮抑制素治疗,分析两组治疗效果。结果治疗结束后,联合组的总有效率72.00高于化疗组的56.00%,差异有统计学意义(χ^(2)=4.167,P<0.05);联合组血清细胞角蛋白19片段、糖类抗原125、癌胚抗原均低于化疗组,差异有统计学意义(P均<0.05);联合组缺氧诱导因子-1α、血管内皮生长因子及血管生成素2、CD8^(+)均低于化疗组(P均<0.05);联合组CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)高于化疗组,CD8^(+)低于化疗组,差异有统计学意义(P均<0.05);两组不良反应发生率对比,差异无统计学意义(P>0.05)。结论持续静脉泵注重组人血管内皮抑制素联合化疗一线对晚期NSCLC治疗效果显著,可明显降低患者血清肿瘤标志物和血管内皮因子水平,改善机体免疫功能,以增强对体内肿瘤细胞活性的控制,进而提高临床疗效,且不增加不良反应。

参苓白术散联合EGFR-TKIs靶向治疗非小细胞肺癌临床观察

目的 参苓白术散联合人表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)靶向治疗非小细胞肺癌(NSCLC)的临床效果。方法 将患者随机分为对照组(29例)和联合组(30例)。对照组采用EGFR-TKIs靶向治疗,联合组在对照组基础上予口服参苓白术散。治疗9周,比较2组临床疗效、中医证候积分、免疫功能指标及不良反应发生率。结果 联合组临床疗效优于对照组(P<0.05)。治疗后,2组中医证候积分下降,联合组偏低(P<0.05);2组CD4^(+)、CD4^(+)/CD8^(+)比率升高,联合组偏高(P<0.05),CD8^(+)水平则降低,联合组偏低(P<0.05)。联合组不良反应总发生率低于对照组(P<0.05)。结论 参苓白术散联合EGFR-TKIs靶向治疗NSCLC疗效显著,可改善免疫功能,减轻不良反应。

自噬在α粒子辐射诱发人支气管上皮细胞恶性转化中的作用

目的:探讨自噬在α粒子辐射诱发永生化人支气管上皮细胞BEP2D恶性转化中的作用。方法:采用Western blot检测BEP2D细胞、α粒子作用BEP2D后的第24代细胞RH24以及α粒子作用BEP2D后的恶性转化细胞BERP35T-1(经鉴定为肺鳞癌细胞)内自噬相关蛋白LC3B-II、LC3B-I和P62的表达,透射电镜观察细胞内单位面积自噬小体的数量。分别用40和60μmol/L自噬抑制剂氯喹(CQ)及25 pmol/L自噬激活剂雷帕霉素(Rapa)作用BERP35T-1细胞,采用Western blot法检测细胞内LC3B和P62蛋白的表达情况,CCK-8法检测细胞的存活率,Transwell侵袭实验检测细胞的侵袭数,划痕愈合实验检测细胞划痕闭合率。结果:与BEP2D细胞相比,RH24和BERP35T-1细胞内LC3B-II/I蛋白比值增高(P<0.05或P<0.01),P62蛋白表达降低,自噬小体数量增多(P<0.01)。40和60μmol/L的CQ分别作用BERP35T-1细胞48 h后,细胞内LC3B-II/I蛋白比值和P62蛋白表达水平均升高,细胞的存活率、侵袭数和划痕闭合率均降低(均为P<0.01);25 pmol/L的Rapa作用BERP35T-1细胞48 h后,细胞内LC3B-II/I蛋白比值升高,P62蛋白表达减弱,细胞的存活率、侵袭数和划痕闭合率均升高(P<0.05或P<0.01)。结论:在α粒子辐射诱发BEP2D细胞恶性转化过程中,细胞自噬增强,可能由此提高细胞的增殖、侵袭和迁移能力,从而促进细胞恶性转化。

马钱苷抑制NF-κB通路对A549肺癌细胞增殖、凋亡、侵袭的影响

目的 探讨马钱苷对人非小细胞肺癌A549细胞增殖、凋亡和侵袭的影响及其作用机制。方法 不同浓度马钱苷处理A549细胞后,CCK-8实验检测细胞增殖抑制作用及半数抑制率(IC50);将A549细胞分为对照组、马钱苷低剂量组、马钱苷中剂量组、马钱苷高剂量组和Diprovocim组,显微镜下观察A549细胞形态,细胞划痕实验检测迁移能力,Transwell实验测定细胞的侵袭能力,流式细胞术检测细胞凋亡,Western Blot技术验证环氧合酶(COX)-2、基质金属蛋白酶(MMP)-2、MMP-9、磷酸化(p)-p65、p65蛋白表达。结果 马钱苷抑制A549细胞的增殖(P<0.05);与对照组相比,马钱苷低、中、高剂量组可见悬浮细胞、大部分细胞脱落、细胞皱缩变圆且丝状伪足减少,细胞迁移和侵袭能力以及COX-2、MMP-2、MMP-9、p-p65/p65蛋白表达水平显著下降,凋亡率显著提高(P<0.05);与马钱苷高剂量组相比,Diprovocim组细胞生长状态良好,细胞迁移和侵袭能力以及COX-2、MMP-2、MMP-9、p-p65/p65蛋白表达水平显著提高,凋亡率显著降低(P<0.05)。结论 马钱苷通过抑制核因子-κB通路抑制A549细胞的增殖和侵袭能力,促进细胞凋亡。