The combination of antitumor drugs,exemestane and erlotinib,induced resistance mechanism in H358 and A549 non-small cell lung cancer (NSCLC) cell lines

Context: Estrogens in non-small-cell lung cancer (NSCLC) are important, and their interaction with epidermal growth factor receptor (EGFR) might be crucial.

Objective: This study investigates the effect of exemestane, an aromatase inhibitor, and erlotinib, an EGFR inhibitor, on human NSCLC cell lines; H23, H358 and A549.

Materials and methods: A cell proliferation assay was used for measuring cell number, apoptosis assay for detecting apoptosis and necrosis and immunoblotting for beclin-1 and Bcl-2 proteins detection. An immunofluorescence assay was used for EGFR localization. A migration assay and zymography were used for cell motility and metalloproteinases (MMPs) expression, respectively.

Results: Exemestane, erlotinib or their combination decreased cell proliferation and increased apoptosis. Exemestane’s half maximal inhibitory concentration (IC₅₀) was 50?μM for H23 and H358 cells and 20?μM for A549. The IC₅₀ of erlotinib was 25?μM for all cell lines. Apoptosis increase induced by exemestane was 58.0 (H23), 186.3 (H358) and 34.7% (A549) and by erlotinib was 16.7 (H23), 65.3 (H358) and 66.3% (A549). A synergy effect was observed only in H23 cells. Noteworthy, the combination of exemestane and erlotinib decreased beclin-1 protein levels (32.3?±?19.2%), an indicator of autophagy, in H23 cells. The combination of exemestane and erlotinib partially reversed the EGFR translocation to mitochondria and decreased MMP levels and migration.

Discussion and conclusions: The benefit from a dual targeting of aromatase and EGFR seems to be regulated by NSCLC cell content. The diverse responses of cells to agents might be influenced by the dominance of certain molecular pathways.     

Phase II study of mitonafide in non-small cell lung cancer (NSCLC)

Summary Background: The new intercalative agent Mitonafide was shown in early clinical trials to be toxic to the central nervous system when administered as a short intravenous infusion, but not when given as a 120-hour continuous infusion. Thus, clinical development in different tumor types was pursued using only this administration schedule, Patients and methods: Forty-nine patients with previously untreated non-small cell lung cancer(NSGLC) and at least one measurable site received Mitonafide as a 120-hour continuous (5 days) infusion every 3 weeks. The starting dose was 170 mg/m²/day × 5 in the first 26 patients and 200 mg/m²/day × 5 in the remainder. Patients were evaluated for toxicity after each course and for response every two courses and remained on treatment until excessive toxicity or disease progression were observed. A special test, the Mini-mental state, was used to assess patients' cognitive functions. Results: Of the 49 patients entered, 42 were evaluable for response and toxicity. Toxicity consisted mainly of myelosuppression and no neurologic side effects were observed. Only one patient presented a partial response. Conclusions: Although definitively safe with this schedule of administration, Mitonafide is not active in NSCLC.     

Targeted delivery of quercetin by biotinylated mixed micelles for non-small cell lung cancer treatment

Lung cancer is the leading cause of cancer death world-wide and its treatment remains a challenge in clinic, especially for non-small cell lung cancer (NSCLC). Thus, more effective therapeutic strategies are required for NSCLC treatment. Quercetin (Que) as a natural flavonoid compound has gained increasing interests due to its anticancer activity. However, poor water solubility, low bioavailability, short half-life, and weak tumor accumulation hinder in vivo applications and antitumor effects of Que. In this study, we developed Que-loaded mixed micelles (Que-MMICs) assembled from 1,2-distearoyl-sn-glycero-3-phosphoethanolamine–poly(ethylene glycol)–biotin (DSPE–PEG–biotin) and poly(ethylene glycol) methyl ether methacrylate–poly[2-(dimethylamino) ethyl acrylate]–polycaprolactone (PEGMA–PDMAEA–PCL) for NSCLC treatment. The results showed that Que was efficiently encapsulated into the mixed micelles and the encapsulation efficiency (EE) was up to 85.7%. Cellular uptake results showed that biotin conjugation significantly improved 1.2-fold internalization of the carrier compared to that of non-targeted mixed micelles. In vitro results demonstrated that Que-MMICs could improve cytotoxicity (IC₅₀ = 7.83 μg/mL) than Que-MICs (16.15 μg/mL) and free Que (44.22 μg/mL) to A549 cells, which efficiently induced apoptosis and arrested cell cycle. Furthermore, Que-MMICs showed satisfactory tumor targeting capability and antitumor efficacy possibly due to the combination of enhanced permeability and retention (EPR) and active targeting effect. Collectively, Que-MMICs demonstrated high accumulation at tumor site and exhibited superior anticancer activity in NSCLC bearing mice model.     

非小细胞肺癌患者紫杉醇3周方案给药后关键药代动力学参数Tc>0.05与临床毒性相关性研究

目的:对接受紫杉醇联合顺铂3周方案的非小细胞肺癌患者药代动力学差异进行研究,分析非小细胞肺癌患者中紫杉醇关键药代动力学参数Tc>0.05(血浓度0.05μM以上时间)与临床毒性的相关性,为紫杉醇的治疗药物监测提供依据。方法:应用纳米增强免疫比浊法检测非小细胞肺癌患者紫杉醇化疗后(24±6)h的血药浓度,采用群体药物动力学模型计算出Tc>0.05值,同时观察化疗后出现的不良反应,按照NCI-CTC v4.0毒副反应评价标准进行分级、结果:全组16例患者共进行32次化疗,有效周期29次(可测算出Tc>0.05),共发生不良反应25例次,包括骨髓抑制、胃肠道反应,无因不良反应而中断治疗者。将患者按照紫杉醇在体内暴露情况分为两组:正常暴露组和高暴露组。骨髓抑制的发生率在两组中存在显著差异(P=0.003),恶心呕吐的发生率无显著性差异(P=0.483)。对两组Ⅲ度以上骨髓抑制的发生率进行比较,发现两组间存在差异,但无统计学意义(P=0.052)。结论:通过测算紫杉醇Tc>0.05值可以了解紫杉醇在患者体内的暴露情况,从而判断患者发生骨髓抑制的风险。但由于本研究中样本量有限,尚需大样本的临床研究进一步证实上述结论。     

CD44v6在NSCLC组织中的表达及意义

目的:研究粘附分子(CD44v6)在非小细胞肺癌中的表达及其临床意义。方法:应用免疫组化(IHC)染色技术检测101例非小细胞肺癌及20例肺良性病变组织CD44v6的表达水平并结合临床资料进行综合分析。结果:在正常肺组织中CD44v6阳性率均在15%,经χ2检验,CD44v6在NSCLC组织和良性病变组织中,两组对比,差异具有显著性(P<0.01)。CD44v6的表达与组织学类型无关(P>0.05),CD44v6表达水平与NSCLC的TNM分期呈正相关,而与NSCLC组织分化程度无关,与淋巴结转移成正相关,CD44v6与原发灶大小无关,CD44v6的阳性表达与术后生存时间呈负相关(P<0.01)。结论:CD44v6在NSCLC中的阳性表达可在一定程度上预示NSCLC具有较强的侵袭和转移能力,CD44v6可作为预测NSCLC患者术后复发转移情况及临床预后的生物学参考指标。     

抗雌激素治疗对耐酪氨酸激酶抑制剂非小细胞肺癌细胞株抗增殖作用的机制探讨

目的 探讨氟维司群逆转吉非替尼耐药非小细胞肺癌细胞株耐药性的可能性及其机制.方法 分别用不同浓度的氟维司群和吉非替尼,单药以及联合对非小细胞肺癌细胞株H1975[含表皮生长因子受体(EGFR)L858R&T790m突变]、H1650(含EGFR Del E746-A750&PTEN De突变)、PC-9(含EGFR Del E746-A750突变)细胞进行干预后,采用MTT法检测细胞增殖变化,Western blot法检测EGFR、雌激素受体(ER)、磷酸化表皮生长因子受体(p-EGFR)的蛋白表达.结果①H1975、H1650、PC-9肺癌细胞中均有EGFR及ER表达;②吉非替尼及氟维司群联合用药较单药可明显抑制H1975、H1650、PC-9肺癌细胞的增殖(P<0.001);③H1975耐药细胞株内T790m突变型EGFR的磷酸化水平可以快速地被雌激素升高或被氟维司群抑制;④ 在酪氨酸激酶抑制剂(EGFR-TKI)获得性耐药的肺癌细胞株中雌激素、表皮生长因子(EGF)分别下调EGFR、ER的水平,氟维司群、吉非替尼分别上调EGFR、ER的水平.结论 采用EGFR的靶向治疗与抗雌激素治疗相结合的治疗方案提高EGFR和ER阳性的EGFR-TKI获得性耐药的NSCLC的治疗效果在理论上是可行的.     

晚期非小细胞肺癌3种化疗方案的成本--效果分析

目的从药物经济学角度,探讨比较不同治疗晚期非小细胞肺癌(NSCLC)的化疗方案的合理性,为临床治疗决策提供依据。方法运用药物经济学方法,对文献报道的MIP、TP、DP3种肺癌化疗方案进行成本-效果分析。结果MIP、TP、DP3种方案的C/E有效率(C/E生存期)分别为132.3(526.9)、734.2(2669.7)和576.4(2619.9)。结论从药物经济学的观点分析,3种方案中MIP方案费用效果比最佳,而TP方案费用最高,疗效最差,费用效果比最差。     

Chalcone-templated Hsp90 inhibitors and their effects on gefitinib resistance in non-small cell lung cancer (NSCLC)

The molecular chaperone Hsp90 has emerged as an attractive cancer therapeutic target due to its role in cellular homeostasis by modulating the stabilization and maturation of many oncogenic proteins. In this study, we designed and synthesized a series of Hsp90 inhibitors that hybridized NVP-AUY992 (2) and PU3 (3) in the chalcone scaffold using a structure-based approach. Our results indicate that compound 1g inhibited the proliferation of gefitinib-resistant non-small cell lung cancer (H1975) cells, downregulated the expression of client proteins of Hsp90 including EGFR, MET, Her2 and Akt, and up-regulated the expression of Hsp70. The compound 1g represents a new class of Hsp90 inhibitors with a chalcone structure. The design, synthesis, and evaluation of 1g are described herein.     

维生素C联合同步放化疗治疗III期非小细胞肺癌的临床疗效

目的 探究维生素 C联合同步放化疗（CCRT）对 III期非小细胞肺癌（NSCLC）患者近期疗效、生存质量及免疫球蛋白（IgA、IgM、IgG）和炎症因子水平的影响。方法 将2019年3月—2021年3月贵州省黔西南布依族苗族自治州人民医院收治的 60例 III期 NSCLC患者随机分为对照组（n＝30）与试验组（n＝30）。对照组患者采用 CCRT治疗。放疗方案：6MV-X 直线加速器扫描,每次 1.8～2.0 Gy,每周 5 次,总剂量 60～66 Gy。化疗方案：顺铂（50 mg·m^-2）第 1 天（d1）、d8、d29、d36静脉滴注,依托泊苷（50 mg·m^-2）d1～d5、d29～d33静脉滴注。36 d为 1个疗程,共治疗 1个疗程。试验组患者采用维生素 C联合 CCRT治疗,CRRT方案同对照组,CCRT治疗方案开始后即给予静脉输注维生素 C,每天 10 g,每周2次（间隔3 d）至放化疗结束,放化疗全程口服维生素C,每天4 g,连续治疗36 d。观察两组患者治疗后近期疗效、生存质量（KPS 评分）以及血清肿瘤坏死因子-α（TNF-α）、IgA、C 反应蛋白（CRP）、IgM、白细胞介素 6（IL-6）、IgG 水平。结果 试验组客观缓解率为 53.33%,略高于对照组的 46.67%,但差异无显著性（P＞0.05）。试验组化疗后生存质量稳定改善率 83.33% 显著高于对照组 56.67%（P＜0.05）。试验组总不良反应发生率 33.33%,显著低于对照组的 60.00%（P＜0.05）。治疗前,两组的 IgA、IgG、IgM 和 IL-6、CRP、TNF-α 水平比较,差异无统计学意义（P＞0.05）;治疗后,两组IgA、IgG、IgM水平均较本组治疗前显著升高（P＜0.05）,IL-6、CRP、TNF-α水平较本组治疗前降低（P＜0.05）;与对照组治疗后比较,试验组 IgA、IgG、IgM 水平显著升高（P＜0.05）,IL-6、CRP、TNF-α水平降低（P＜0.05）。结论 维生素C联合CCRT治疗III期NSCLC可显著降低不良反应发生率,改善生存质量及机体免疫功能。     

Antiproliferative activity of parthenolide against three human cancer cell lines and human umbilical vein endothelial cells

Parthenolide is a major sesquiterpene lactone derived from feverfew (Tanacetum parthenium) with known anti-inflammatory activity. Moreover, the anticancer potential of this compound was suggested. In this study, we determined the effect of parthenolide on proliferation of three human cancer cell lines: human lung carcinoma (A549), human medulloblastoma (TE671), human colon adenocarcinoma (HT-29) and human umbilical vein endothelial cells (HUVEC) in vitro. Cell proliferation was assessed by means of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The IC(50) value (the concentration of drug necessary to induce 50% inhibition) together with confidence limits was calculated. Parthenolide inhibited proliferation of all three types of cancer cells (A549, TE671, HT-29) and HUVEC with the following IC(50) values (in muM): 4.3, 6.5, 7.0 and 2.8, respectively. Thus, the antiproliferative potential of parthenolide was confirmed.     

安罗替尼联合白蛋白结合型紫杉醇三线治疗非小细胞肺癌的临床疗效和安全性#br#

目的　探讨安罗替尼联合白蛋白结合型紫杉醇（Nab-P）三线治疗非小细胞肺癌（NSCLC）患者的疗效和安全性。方法　选取79例NSCLC患者作为研究对象,按照治疗方案分为对照组（36例）和研究组（43例）,对照组采用Nab-P治疗,研究组采用安罗替尼联合Nab-P治疗。观察2组临床疗效,随访记录患者无进展生存期（PFS）、总生存期（OS）及不良反应;同时观察2组治疗前后血清血管内皮生长因子（VEGF）、癌胚抗原（CEA）和糖类抗原125（CA125）水平变化情况。结果　2组客观缓解率差异无统计学意义（16.28% vs. 8.33%,P＞0.05）;研究组疾病控制率高于对照组（76.74% vs. 52.78%,P＜0.05）。2组中位PFS分别为4个月和3个月（P＞0.05）;中位OS分别为9个月和7个月（P＜0.05）。治疗后,2组血清VEGF、CEA和CA125水平较治疗前均降低,且研究组低于对照组（P＜0.05）。治疗期间,2组不良反应发生率差异无统计学意义（P＞0.05）。结论　安罗替尼联合Nab-P三线治疗NSCLC具有较好的疗效,可有效降低血清VEGF、CEA和CA125水平,且安全性较好。     

Dose-finding study of paclitaxel and carboplatin in patients with advanced non-small cell lung cancer

This dose-finding study was designed to determine the maximum tolerated dose (MTD), efficacy and toxicity of combined paclitaxel and carboplatin in 35 previously untreated patients with advanced non-small cell lung cancer (NSCLC). Paclitaxel was given as a 3-h infusion at escalating dose levels (100-250 mg/m2) immediately followed by carboplatin as a 30-min infusion (325 or 350 mg/m2) every 3 weeks. The dose-limiting toxicity, paresthesia, occurred at the highest dose level, therefore the recommended dose was established one level below (paclitaxel 225 mg/m2 with carboplatin 325 mg/m2). Neutropenia was the most common hematotoxicity; dose dependency was not apparent. Two patients, at different dose levels, had febrile neutropenia. Thrombocytopenia was rare. Non-hematological toxicities grade 3 or higher included infection, anorexia, alopecia and paresthesia. One patient had a hypersensitivity reaction (transient hypotension). The overall response rate was 23% and median survival time was 7.5 months. Promising activity and acceptable toxicity supports the development of this combination as a useful chemotherapeutic option in advanced NSCLC.     

紫杉醇脂质体与紫杉醇治疗老年非小细胞肺癌的疗效观察

目的比较紫杉醇脂质体与普通紫杉醇治疗老年非小细胞肺癌的疗效及毒副反应。方法 68例老年非小细胞肺癌患者随机分为试验组和对照组。试验组33例给予紫杉醇脂质体135mg/m2,对照组35例给予紫杉醇135mg/m2,每21天为一周期,共2周期。观察两组的疗效及毒副反应情况。结果两组在疗效及血液学毒性、消化道反应的差异无统计学意义,但因溶媒产生的过敏反应方面,试验组明显低于对照组,有统计学意义。结论紫杉醇脂质体治疗老年非小细胞肺癌效果良好,两种紫杉醇疗效相当,但紫杉醇脂质体的过敏反应明显低于紫杉醇。     

组织蛋白酶-D和MMP-9在非小细胞肺癌中的表达意义

目的检测组织蛋白酶-D（Cath-D）和基质金属蛋白酶-9（MMP-9）在非小细胞肺癌中的表达,探讨Cath-D和MMP-9在非小细胞肺癌发生发展及转移中的作用。方法应用免疫组织化学法检测96例非小细胞肺癌组织及60例正常肺组织中Cath-D和MMP-9的表达情况,并将检测结果与临床病理特征进行综合分析。结果 Cath-D和MMP-9的表达主要定位于细胞质,Cath-D和MMP-9在非小细胞肺癌中表达的阳性率明显高于正常肺组织。非小细胞肺癌中Cath-D和MMP-9蛋白的表达与有无胸膜侵犯和淋巴结转移情况密切相关。结论 Cath-D和MMP-9在非小细胞肺癌中表达上调,联合检测Cath-D和MMP-9可能对判断肿瘤的预后有重要价值。     

小白菊内酯对人非小细胞性肺癌H1975细胞凋亡、侵袭与迁移的影响

目的探究小白菊内酯(parthenolide, PTL)对非小细胞性肺癌细胞株H1975凋亡、侵袭和迁移的影响及其可能机制。方法 MTT法和集落克隆实验检测PTL对H1975细胞增殖的影响;Annexin V-FITC/PI双染、流式细胞仪检测PTL对H1975细胞凋亡的影响;Transwell实验检测PTL对H1975细胞侵袭和迁移的影响;Western blot检测细胞凋亡、侵袭和迁移相关蛋白,以及PI3K/Akt信号通路相关蛋白的表达。结果 MTT和集落克隆实验结果显示,随着PTL浓度的增加,H1975细胞的增殖明显受到抑制,与对照组相比,差异均具有统计学意义(P<0.05);Annexin V-FITC/PI双染结果显示,PTL能明显诱导H1975细胞发生凋亡(P<0.01);Transwell实验结果显示,PTL能明显抑制H1975细胞侵袭和迁移(P<0.01);Western blot结果显示,PTL明显上调H1975细胞Bax蛋白表达,下调Bcl-2、HIF-1α、MMP-9、Akt、p-Akt(Ser473)蛋白的表达(P<0.05),同时使caspase-3蛋白发生裂解。结论 PTL能明显诱导非小细胞性肺癌H1975细胞发生凋亡,抑制其侵袭和迁移,作用机制可能与PTL抑制PI3K/Akt信号通路有关。     

Cremophor EL pharmacokinetics in a phase I study of paclitaxel (Taxol) and carboplatin in non-small cell lung cancer patients

The purpose of our study was to investigate the pharmacokinetics of Cremophor EL following administration of escalating doses of Taxol (paclitaxel dissolved in Cremophor EL/ethanol) to non-small cell lung cancer (NSCLC) patients. Patients with NSCLC stage IIIb or IV without prior chemotherapy treatment were eligible for treatment with paclitaxel and carboplatin in a dose-finding phase I study. The starting dose of paclitaxel was 100 mg/m2 and doses were escalated with steps of 25 mg/m2, which is equal to a starting dose of Cremophor EL of 8.3 ml/m2 with dose increments of 2.1 ml/m2. Carboplatin dosages were 300, 350 or 400 mg/m2. Pharmacokinetic sampling was performed during the first and the second course, and the samples were analyzed using a validated high-performance liquid chromatographic assay. A total of 39 patients were included in this pharmacokinetic part of the study. The doses of paclitaxel were escalated up to 250 mg/m2 (20.8 ml/m2 Cremophor EL). Pharmacokinetic analyses revealed a low elimination-rate of Cremophor EL (CI=37.8-134 ml/h/m2; t 1/2=34.4-61.5 h) and a volume of distribution similar to the volume of the central blood compartment (Vss=4.96-7.85 l). In addition, a dose-independent clearance of Cremophor EL was found indicating linear kinetics. Dose adjustment using the body surface area, however, resulted in a non-linear increase in systemic exposure. The use of body surface area in calculations of Cremophor EL should therefore be re-evaluated.     

Hyaluronic acid-modified didecyldimethylammonium bromide/ d-a-tocopheryl polyethylene glycol succinate mixed micelles for delivery of baohuoside I against non-small cell lung cancer: in vitro and in vivo evaluation

Baohuoside I is an effective but a poorly soluble antitumor drug. In this study, we prepared baohuoside I-loaded mixed micelles with didecyldimethylammonium bromide (DDAB) and d-a-tocopheryl polyethylene glycol succinate (TPGS) (DTBM) and active targeting mixed micelles (HDTBM) with hyaluronic acid (HA) as the targeting ligand on the surface of the mixed micelles. We performed a systematic comparative evaluation of the antiproliferative effect, cellular uptake, antitumor efficacy, and in vivo tumor targeting of these micelles using A549 cells. HDTBM showed improved cellular uptake and had a greater hypersensitizing effect on A549 cell lines than baohuoside I; half-maximal inhibitory concentration (IC₅₀) was 8.86 versus 20.42?μg/mL, respectively. Results of the antitumor efficacy study and the imaging study for in vivo targeting showed that the mixed-micelle formulation had higher antitumor efficacy and achieved effective and targeted drug delivery. Therefore, our results indicate that HA/baohuoside I-M may be used as a potential antitumor formulation.