Retracted Article: Structural characterization of Momordica charantia L. (Cucurbitaceae) oligopeptides and the detection of their capability in non-small cell lung cancer A549 cells: induction of apoptosis

Oligopeptides are rarely reported from Chinese herbal medicine because they are often present in very low concentrations in a complex matrix. Twenty-eight oligopeptides were recently identified by high-performance liquid chromatography and quadrupole-time-of-flight-mass spectrometry (HPLC-Q-TOF-MS) from Momordica charantia L. (Cucurbitaceae), and a septapeptide, FHGKGHE (Phe-His-Gly-Lys-Gly-His-Glu), named MCLO-12, showed the best anticancer activity against the non-small cell lung cancer A549 cell line in vitro, with an IC₅₀ value of 21.4 ± 2.21 mM. The anti-proliferative activity assay results showed that MCLO-12 induced apoptosis of A549 cells in a concentration-dependent manner. Treatment of the cells with MCLO-12 (10.7–42.8 mM mL⁻¹) caused strong intracellular reactive oxygen species (ROS) up-regulating activities and activated caspase expression. MCLO-12 also suppressed the Trx system and subsequently activated a number of Trx-dependent pathways, including the ASK1, MAPK-p38 and JNK pathways. Thus, our research provides a good reference point for anti-NSCLC research into oligopeptides.

MCLO-12 induced apoptosis by up-regulating the ROS, activating the caspases expressions, suppressing the Trx system and subsequently activating a number of Trx-dependent pathways.     

Retraction: Structural characterization of centipede oligopeptides and capability detection in human small cell lung carcinoma: inducing apoptosis

Retraction of ‘Structural characterization of centipede oligopeptides and capability detection in human small cell lung carcinoma: inducing apoptosis’ by JingQuan Zhao et al., RSC Adv., 2019, 9, 10927–10936, https://doi.org/10.1039/C8RA09018A.

The Royal Society of Chemistry hereby wholly retracts this RSC Advances article due to a significant amount of unattributed text overlap throughout the article, and particularly with ref. 1 in the Results and discussion section and ref. 2 in the Conclusion section.Jie Liu opposes the retraction. The other authors have been informed but have not responded to any correspondence regarding the retraction.Signed: Laura Fisher, Executive Editor, RSC AdvancesDate: 29^th March 2022     

Retracted Article: Structural characterization of centipede oligopeptides and capability detection in human small cell lung carcinoma: inducing apoptosis

Lung cancer is the most frequent cause of cancer deaths in the world, and smoking is considered as one of the major causes. Small cell lung carcinoma (SCLC) represents a highly malignant and particularly aggressive form, with properties of widespread metastases and poor prognosis. Herein, twenty-five Scolopendra subspinipes mutilans L. Koch Oligopeptides (SSMOs) were isolated and their structures were identified, and the anti-proliferative activity against lung cancer cell lines was evaluated. Results showed that SSMO-5 induced the production of reactive oxygen species (ROS) markedly in NCI-H446 cells. Furthermore, SSMO-5 decreased the mitochondrial membrane potential (MMP) and enhanced the mitochondria-related apoptosis. These results demonstrate that in NCI-H446 cells, the apoptotic and cytotoxic effects of SSMO-5 are mediated by the intrinsic mitochondria-mediated apoptotic pathway, which in turn causes the activation of caspases and increases Bax expression, while decreases Bcl-2 and Bcl-xL expressions and regulates the interaction of p53/MDM2. In conclusion, a ROS-mediated mitochondrial pathway plays an important role in the process of SSMO-5-induced apoptosis against SCLC.

SSMO-5 mediated the lung cancer cells apoptosis by activating the caspases and regulating the interaction of p53/MDM2.     

Vinculin表达对非小细胞肺癌A549细胞生物学特征的影响

目的通过体外实验探讨非小细胞肺癌(NSCLC)A549细胞系的Vinculin表达对其生物学特征的影响。方法购买人NSCLC A549细胞系,体外培养,分为对照组、空白载体组以及高表达组三组,高表达组A549细胞系转染Vinculin过表达载体,空白载体组A549细胞系转染空白载体。体外培养48 h后,应用RT-PCR检测各组A549细胞系Vinculin mRNA表达情况,应用MTT法检测细胞活性,Ki-67免疫荧光检测细胞增殖能力,Transwell培养体系Hoechst染色观察A549细胞系迁移侵袭能力。结果 RT-PCR检测结果显示,与对照组、空白载体组比较,高表达组Vinculin mRNA表达量明显增高(P<0.01);MTT法检测结果显示,与对照组、空白载体组比较,高表达组的OD值明显降低(P<0.01);Ki-67免疫荧光检测结果显示,与对照组、空白载体组比较,高表达组Ki-67⁺细胞数量明显减少(P<0.01);Transwell培养体系Hoeschst染色结果显示,与对照组、空白载体组比较,高表达组迁移和侵袭的细胞数量明显减少...     

Retracted Article: Structural characterization of ginseng cyclopeptides and detection of capability to induce apoptosis in gastrointestinal cancer cells

Gastrointestinal tumors are the most frequently diagnosed malignancy and the second highest contributor to cancer mortality. Cyclopeptides are rarely isolated from ginseng because they are often present at low concentrations in a complex matrix. In the current study, seven novel ginseng cyclopeptides (GCPs) were isolated and their anti-tumor potency was explored. Anti-proliferative test results show that the (GCP-1)∼[cyclo-(_L-Trp-_L-Glu-_L-Phe-_L-Thr)] peptide display the best anti-proliferative activity in gastric cancer SGC-7901 cells in vitro, with an IC₅₀ value of 37.8 ± 3.13 μM. Flow cytometry analysis shows that GCP-1 (7.56–189 μM) clearly induce early apoptosis and mitochondrial membrane potential collapse, and block the cells at the G0/G1 phase. A further study revealed that GCP-1 induces apoptosis by activating the caspases, suppressing the thioredoxin (Trx) system and subsequently activating a number of Trx-dependent pathways, including those involving apoptotic signal-regulating kinase 1 (ASK1) and mitogen-activated protein kinases (MAPKs). The cyclopeptides in ginseng are an important resource for the research and development of anti-neoplastic drugs.

Gastrointestinal tumors are the most frequently diagnosed malignancy and the second highest contributor to cancer mortality.     

Structural characterization and anticancer potency of centipede oligopeptides in human chondrosarcoma cancer: inducing apoptosis

Anticancer oligopeptides are rarely studied because they are often present in very low concentrations in a complex matrix. In the current study, twelve oligopeptides were isolated and the amino acid sequence identified from the centipede. MTT results indicated that Trp–Gly–His–Glu (CO-10) showed excellent anti-proliferative potency against chondrosarcoma cells in vitro. Further study showed that CO-10 induced SW1353 cells apoptosis and blocked cell cycle in the G0/G1 phase. Further, results demonstrate that the apoptotic and cytotoxic effects of CO-10 are mediated by the intrinsic mitochondria-mediated apoptotic pathway, which in turn causes the release of cytochrome c and the activation of caspases. This study will be important for the development of pharmaceutical anticancer peptides from natural products as anticancer agents against chondrosarcoma.

CO-10 induced cell apoptosis with mitochondria dysfunction and caspases, ASK1-MAPKs pathway activation.     

帕瑞昔布对非小细胞肺癌细胞株A549增殖和迁移的影响

目的 研究帕瑞昔布对非小细胞肺癌A549细胞增殖和迁移的影响,并探讨其可能机制.方法 采用随机数字表法将A549细胞随机分为四组：对照组（C组）,10 μmol/L帕瑞昔布（P1组）、40 μmol/L帕瑞昔布组（P2组）和160 μmol/L帕瑞昔布组（P3组）.C组细胞常规培养,P1、P2和P3组细胞分别用终浓度为10 μmol/L、40 μmol/L和160 μmol/L的帕瑞昔布处理A549细胞24 h.MTT法检测各组细胞的增殖情况,划痕试验检测各组细胞的迁移能力,Western blot法检测各组细胞p-AKT、survivin的表达情况.结果 与C组比较,P1、P2和P3组细胞的增殖抑制率依次增高（P＜0.05）,P1、P2和P3组细胞的迁移距离依次降低（P＜0.05）,P1、P2和P3组细胞p-AKT和survivin的表达水平依次降低（P＜0.05）,且呈剂量依赖性.结论 帕瑞昔布可以抑制A549细胞的增殖和迁移,其机制可能是抑制p-AKT和survivin的表达.     

IRE1α-TRAF2-ASK1 pathway is involved in CSTMP-induced apoptosis and ER stress in human non-small cell lung cancer A549 cells

BackgroundCSTMP, a Tetramethylpyrazine (TMP) analogue, is designed and synthesized based on the pharmacophores of TMP and resveratrol. Recent studies showed that CSTMP had strong protective effects in endothelial cells apoptosis by its anti-oxidant activity. However, the pharmacological function of CSTMP in cancer have not been elucidated to date. The objective of this study was to investigate the anti-cancer effect of CSTMP against human non-small cell lung cancer (NSCLC) A549 cells and the underlying mechanisms.MethodsThe cell proliferation and apoptosis were detected by MTT assay and flow cytometry. Caspases activity was determined spectrophotometricaly at 405 nm using a microtiter plate reader. Western blot and real-time PCR was used to assess the protein and mRNA expression. Immunoprecipitation was used to examine the protein–protein interactions.ResultsCSTMP inhibited the proliferation and induced cell cycle arrest and apoptosis of A549 cells. Caspase3, 8, 9 and PARP-1 activation, and Bax/Bcl-2 ratio analyses demonstrated that the anti-cancer effect of CSTMP in A549 cells was mediated by promoting caspase- and mitochondria-dependent apoptosis. Furthermore, CSTMP induced ER stress in A549 cells as evidenced by elevated levels of GRP78, GRP94, CHOP, IRE1α, TRAF2, p-ASK1 and p-JNK, activation of caspase12 and 4, and enhanced formation of an IRE1α-TRAF2-ASK1 complex. Knockdown of IRE1α by siRNA suppressed activation of IRE1α, TRAF2, p-ASK1 and p-JNK in CSTMP treated A549 cells. In addition, the effects of CSTMP on the formation of an IRE1α-TRAF2-ASK1 complex, caspase- and mitochondria-dependent apoptosis were also reversed by IRE1α siRNA in A549 cells.ConclusionsCollectively, we showed that CSTMP induced apoptosis of A549 cells were through IRE1α-TRAF2-ASK1 complex-mediated ER stress, JNK activation, and mitochondrial dysfunction. These insights on this novel compound CSTMP may provide a novel anti-cancer candidate for the treatment of NSCLC.     

新藤黄酸对人肺癌细胞株A549的增值和凋亡的影响

目的探讨新藤黄酸(GNA)对人肺癌细胞株A549的增值和凋亡的影响及其调控的可能机制。方法用不同浓度的GNA处理A549细胞,分为对照组和GNA组(GNA浓度分别为0.5、1.5、2.0mg/L),采用MTT、细胞划痕、Hoechst染色法观察细胞的增殖、迁移和凋亡状况;Western blot检测凋亡相关蛋白caspase3、caspase9、p53和NF-κB的表达变化。结果 GNA能明显抑制细胞的体外增殖和迁移且呈剂量依赖性;Hoechst染色显示GNA能诱导细胞凋亡,且随着浓度的升高凋亡现象越来越明显;Western blot显示GNA能上调促进凋亡的蛋白caspase3、caspase9、p53和下调NF-κB的表达(P0.05)。结论 GNA能明显抑制人肺癌细胞株A549的增殖和迁移并诱导凋亡。     

Afatinib triggers a Ni2+-resistant Ca2+ influx pathway in A549 non-small cell lung cancer cells

Afatinib is used to treat non-small cell lung cancer cells (NSCLC), and its mechanism involves irreversible inhibition of epidermal growth factor receptor (EGFR) tyrosine kinase. In this study, we examined if afatinib had cytotoxic action against NSCLC other than inhibition of tyrosine kinase. Afatinib (1–30 μM) caused apoptotic death in A549 NSCLC in a concentration-dependent manner. Afatinib triggered Ca²⁺ influx without causing Ca²⁺ release, and the Ca²⁺ influx was unaffected by sodium orthovanadate (SOV, an inhibitor of tyrosine phosphatase), suggesting that afatinib-triggered Ca²⁺ response was unrelated to its inhibition of tyrosine kinase. Addition of afatinib also promoted Mn²⁺ influx. Ca²⁺ influx triggered by afatinib was resistant to SKF96365 and ruthenium red (two general blockers of TRP channels) and, unexpectedly, Ni²⁺ (a non-specific Ca²⁺ channel blocker). Afatinib caused an increase in mitochondrial Ca²⁺ level, an initial mitochondrial hyperpolarization (4 h) and followed by mitochondrial potential collapse (24–48 h). Afatinib-induced cell death was slightly but significantly alleviated in low extracellular Ca²⁺ condition or under pharmacological block of mitochondrial permeability transition pore (MPTP) opening by cyclosporin A. Therefore, in addition to tyrosine kinase inhibition as a major anti-cancer mechanism of afatinib, stimulation of an atypical Ca²⁺ influx pathway, mitochondrial Ca²⁺ overload, and potential collapse in part contribute to afatinib-induced cell death.     

直流小电场对肺癌细胞凋亡的诱导作用

目的观察应用直流电疗法对肺癌细胞凋亡的影响,研究直流小电场对于肺癌细胞凋亡的诱导作用及其可能机制。方法应用体外直流小电场作用于肺腺癌细胞系A549,观察其对肺癌细胞凋亡和重要基因p53、Rb以及E2F1表达的影响。结果直流小电场对肺癌细胞系A549有明显的诱导作用,200mV/mm和250mV/mm组癌细胞大量脱落,电场组凋亡癌细胞数目随电场强度升高而增多(F=795.372,P=0.000)。电场组p53、Rb基因mRNA表达明显升高,E2F1基因mRNA表达明显下降。结论直流小电场对肺癌细胞系A549具有明显的凋亡诱导作用并可能与p53、Rb以及E2F1基因的表达相关。     

A humanized anti-IL-6 antibody (ALD518) in non-small cell lung cancer

INTRODUCTION: Inflammatory pathways may be an important contributor to morbidity and mortality associated with lung cancer. The oncogene-associated inflammatory microenvironment leads to production of inflammatory cytokines such as IL-6. IL-6 is associated with poor prognosis and correlates with debilitating lung-cancer-related symptoms such as fatigue, thromboembolism, cachexia and anemia. IL-6 has been implicated in resistance of lung cancer to EGF inhibitors. A mAb therapy targeting IL-6 may be an effective treatment for the inflammatory microenvironment in lung cancer. AREAS COVERED: An understanding of the inflammatory pathways involved in lung cancer, including the central role of IL-6, and how inflammation affects the course and treatment of lung cancer. The mAb ALD518, which targets IL-6, and its investigational development and use in advanced NSCLC. Preclinical and Phase I and II studies of ALD518 with a focus on NSCLC. How ALD518 could be used in NSCLC in the future. EXPERT OPINION: IL-6-mediated inflammation may contribute to NSCLC-related morbidity and mortality. In preclinical and Phase I and II trials ALD518 targeting IL-6 appears well tolerated and ameliorates NSCLC-related anemia and cachexia. Other clinical outcomes need further study, and may include effects on overall survival, hypercoagulability associated with lung cancer and decreased resistance to EGF-pathway inhibitors.     

活性氧在荆花牡荆素诱导人肺癌A549细胞凋亡中的作用

目的 探讨紫花牡荆素(CAS)诱导人肺腺癌A549细胞凋亡及其机制.方法 体外培养A549细胞.MTT法测定CAS对A549细胞增殖的抑制;Annexin V/PI双染色分析细胞凋亡率;H2DCFH-DA探针流式细胞术分析活性氧(ROS)生成.结果 CAS能抑制人肺癌A549细胞增殖,呈浓度依赖性.Annexin V/PI法检测结果显示10 μmol/L的CAS作用A549细胞12 h、24 h、48 h后,其凋亡率分别为22.39%、38.66%、64.82%.H2DCFH-DA探针流式细胞术分析表明,完全培养基组、溶媒(0.1% DMSO)组对A459细胞作用0 h;CAS(10 μmol/L)对A549细胞作用3 h、6 h、12 h;N-乙酰-L-半胱氨酸(NAC,10 mmol/L)+CAS(10 μmol/L)组对A549细胞作用6 h的ROS生成水平分别为8.47、15.26、66.2、74.1、82.2、67.3,随着CAS作用时间延长,细胞内ROS水平增加.NAC对细胞凋亡有抑制作用.结论 CAS可诱导A549细胞凋亡,其作用机制可能与提高细胞内ROS产生增加有关.     

外周血叶酸受体阳性循环肿瘤细胞检测在非小细胞肺癌筛查中的应用价值

目的观察非小细胞肺癌患者外周血叶酸受体阳性循环肿瘤细胞(circulating tumor cells,CTCs)水平变化,探讨外周血叶酸受体阳性CTCs在非小细胞肺癌筛查中的应用价值。方法非小细胞肺癌患者136例为肺癌组,肺部良性病变患者10例为良性病变组,健康志愿者54例为对照组。3组均采用以叶酸受体为靶点的免疫磁珠阴性富集+实时荧光定量PCR法检测外周血叶酸受体阳性CTCs水平;采用化学发光免疫分析法检测血清癌胚抗原(carcino-embryonic antigen,CEA)、糖链抗原(carbohydrate antigen,CA)125、CA724、细胞角蛋白19片段(cytokeratin 19fragment,CYFRA21-1)、神经元特异性烯醇化酶(neuron-specific enolase,NSE)水平。比较3组CTCs水平;比较不同临床特征非小细胞肺癌患者CTCs水平;绘制ROC曲线,评价CTCs及血清CEA、CA125、CA724、CYFRA21-1、NSE 5项指标联合诊断非小细胞肺癌的价值。结果肺癌组CTCs水平[11.21(8.58,15.30)FU/3mL]高于良性病变组[7.55(5.23,10.25)FU/3mL]和对照组[4.95(3.55,7.62)FU/3mL](P<0.05),良性病变组高于对照组(P<0.05)。TNM分期Ⅰ、Ⅱ、Ⅲ、Ⅳ期非小细胞肺癌患者CTCs水平[11.00(8.58,13.30)、13.25(10.48,16.88)、14.77(11.47,16.55)、17.89(17.07,19.22)FU/3mL]两两比较差异均有统计学意义(H=16.443,P<0.05);不同年龄、性别、肿瘤最大径、T分期、分化等级、病理类型非小细胞肺癌患者CTCs水平比较差异均无统计学意义(P>0.05)。ROC曲线分析结果显示,CTCs以8.70FU/3mL为最佳截断值,诊断非小细胞肺癌的AUC为0.953(95%CI:0.926~0.979,P<0.05),灵敏度为79.40%,特异度为98.10%,诊断效能优于CEA、CA125、CA724、CYFRA21-1、NSE联合检测。结论非小细胞肺癌患者外周血叶酸受体阳性CTCs水平升高,且增高程度与TNM分期有关;外周血叶酸受体阳性CTCs可用于非小细胞肺癌的早期筛查。     

Ipilimumab (MDX-010) in the treatment of non-small cell lung cancer

Respiratory syncytial virus (RSV) is a major cause of bronchiolitis and pneumonia in young children and the elderly. Despite its clinical importance, there is no licensed vaccine available at present. Vaccine development has been hampered by observations of increased pathology after RSV infection in infants vaccinated with formalin-inactivated RSV; incomplete immunity following natural infection; and the need to be effective during the neonatal period when levels of maternal antibody are high. Four categories of RSV vaccine carriers – live-attenuated RSVs, recombinant vectors expressing the protective antigens of RSV, DNA vaccines and subunit vaccines – have been evaluated in animal models and/or clinical trials. So far, studies with live-attenuated virus vaccines highlight the need to improve immunogenicity whilst maintaining a suitable level of attenuation. Studies with recombinant vectors, DNA and subunit vaccines illustrate the pivotal nature of the vaccine carrier in determining the balance between immune-mediated protection against infection and the induction of immune-mediated pulmonary pathology.     

Ipilimumab (MDX-010) in the treatment of non-small cell lung cancer

INTRODUCTION: Non-small cell lung cancer (NSCLC) is one of the main causes of cancer-related deaths worldwide. Although new therapies have become available, innovative treatments are still needed for advanced disease. Ipilimumab , a monoclonal antibody targeting cytotoxic T-lymphocyte antigen 4 (CTLA-4), enhances the immune response against the tumor mass and has been proven effective against malignant melanoma. AREAS COVERED: The authors explored the role of ipilimumab in NSCLC using a literature review. The clinical trials involving ipilimumab for lung cancer have shown progression-free survival (PFS) benefits. The use of ipilimumab is related to unusual adverse events resulting from increased or excessive immune activity. Because ipilimumab shows unique response patterns, more suitable criteria known as immune-related response criteria (ir-RC), different from RECIST and WHO criteria, are required. EXPERT OPINION: Although NSCLC is not known as an immunogenic-mediated malignancy, in the past few years, the authors have observed an increasing interest in the development of therapies able to modulate the immune response including vaccines and non-specific immunoregulatory drugs (such as ipilimumab). Ipilimumab may become a new, powerful strategy for the management of NSCLC patients. Further investigation is needed to confirm the optimal treatment schedule and determine the potential predictors of response to the CTLA-4 blockade.     

Detection of the mesenchymal-to-epithelial transition of invasive non-small cell lung cancer cells by their membrane undulation spectra

A cancer cell changes its state from being epithelial- to mesenchymal-like in a dynamic manner during tumor progression. For example, it is well known that mesenchymal-to-epithelial transition (MET) is essential for cancer cells to regain the capability of seeding on and then invading secondary/tertiary regions. However, there is no fast yet reliable method for detecting this transition. Here, we showed that membrane undulation of invasive cancer cells could be used as a novel marker for MET detection, both in invasive model cell lines and repopulated circulating tumor cells (rCTCs) from non-small cell lung cancer (NSCLC) patients. Specifically, using atomic force microscopy (AFM), it was found that the surface oscillation spectra of different cancer cells, after undergoing MET, all exhibited two distinct peaks from 0.001 to 0.007 Hz that are absent in the spectra before MET. In addition, by adopting the long short-term memory (LSTM) based recurrent neural network learning algorithm, we showed that the positions of recorded membrane undulation peaks can be used to predict the occurrence of MET in invasive NSCLC cells with high accuracy (>90% for model cell lines and >80% for rCTCs when benchmarking against the conventional bio-marker vimentin). These findings demonstrate the potential of our approach in achieving rapid MET detection with a much reduced cell sample size as well as quantifying changes in the mesenchymal level of tumor cells.

The membrane undulation spectra of cancer cells, measured by atomic force microscope, can be used to detect their transition from being mesenchymal- to epithelial-like.