sincalide and Carcinoma--Non-Small-Cell-Lung

Epidermal growth factor receptor (EGFR) gene mutations are the most common driver mutations in non-small cell lung cancer (NSCLC). To prolong the survival of the patients, EGFR tyrosine kinase inhibitors (TKIs) resistance in NSCLC is a major challenge that needs to be addressed urgently, and this study focuses on investigating the mechanism of cigarette smoke (CS) induced Gefitinib resistance in NSCLC.. PC-9 and A549 cells were cultured in vitro and treated with 1 µmol/L Gefitinib for 4 h and 10% cigarette smoke extract (CSE) for 48 h. Western blot was used to detect Sirtuin 3 (Sirt3) and superoxide dismutase 2 (SOD2) protein expressions; DCFH-DA probe was used to detect intracellular reactive oxygen species (ROS); CCK-8 kit was used to detect cell activity, and EdU was used to detect cell proliferation ability. Sirt3 overexpression plasmid (OV-Sirt3) was transfected in PC-9 and A549 cells and treated with 1 µmol/L Gefitinib for 4 h and 10% CSE for 48 h after N-acetylcysteine (NAC) action. The expressions of Sirt3 and SOD2 were detected by Western blot; the ROS level in the cells was detected by DCFH-DA probe, and the cell activity was detected by CCK-8.. CSE induced an increase in the 50% inhibitory concentration (IC50) of both PC-9 and A549 cells to Gefitinib (P<0.01) and enhanced the proliferation of PC-9 and A549 cells, suggesting that CS induced Gefitinib resistance in NSCLC. ROS was involved in CSE-induced Gefitinib resistance (P<0.05). CSE induced low expressions of Sirt3 and SOD2 (P<0.01), and Sirt3/SOD2 was associated with poor prognosis in lung cancer patients (P<0.05). OV-Sirt3 in PC-9 and A549 cells reversed CSE-induced Gefitinib resistance (P<0.05) and significantly reduced ROS production. NAC reversed CSE-induced Gefitinib resistance in PC-9 and A549 cells (P<0.05).. The ROS/Sirt3/SOD2 pathway is involved in CS-induced Gefitinib resistance in NSCLC.. 【中文题目：烟草烟雾通过ROS/Sirt3/SOD2通路
诱导NSCLC细胞吉非替尼耐药】 【中文摘要：背景与目的 表皮生长因子受体（epidermal growth factor receptor, EGFR）基因突变是非小细胞肺癌（non-small cell lung cancer, NSCLC）最常见的驱动基因突变。为延长患者生存时间，NSCLC EGFR酪氨酸激酶抑制剂（tyrosine kinase inhibitors, TKIs）耐药是目前急需解决的重大难题。本研究主要探究烟草烟雾（cigarette smoke, CS）诱导NSCLC发生吉非替尼耐药的机制。方法 体外培养PC-9、A549细胞，分别经1 µmol/L吉非替尼处理4 h、10%CS萃取物（CS extract, CSE）处理48 h。Western blot检测沉默蛋白3（Sirtuin 3, Sirt3）、超氧化物歧化酶2（superoxide dismutase 2, SOD2）蛋白表达，使用DCFH-DA探针检测细胞内活性氧（reactive oxygen species, ROS）水平，CCK-8试剂盒检测细胞活性，EdU检测细胞增殖能力。Sirt3过表达质粒（OV-Sirt3）转染于PC-9和A549细胞中、N-乙酰半胱氨酸乙酯（N-acetylcysteine, NAC）作用于细胞后分别经1 µmol/L吉非替尼处理4 h和10%CSE处理48 h。Western blot检测Sirt3、SOD2蛋白表达，DCFH-DA探针检测细胞中的ROS水平，CCK-8检测细胞活性。结果 CSE均可促使PC-9、A549细胞对吉非替尼的半数抑制浓度（50% inhibitory concentration, IC50）提高（P<0.01），并且增强PC-9和A549细胞的增殖能力，提示CS可诱导NSCLC吉非替尼耐药；ROS参与CSE诱导的吉非替尼耐药（P<0.05）；CSE诱导Sirt3、SOD2低表达（P<0.01），且Sirt3/SOD2与肺癌患者不良预后有关（P<0.05）；OV-Sirt3的PC-9、A549细胞可逆转CSE诱导的吉非替尼耐药（P<0.05）且显著降低ROS生成；NAC可逆转CSE诱导的PC-9、A549细胞吉非替尼耐药（P<0.05）。结论 ROS/Sirt3/SOD2通路参与了CS诱导的NSCLC吉非替尼耐药。
】 【中文关键词：肺肿瘤；烟草烟雾；ROS；Sirt3/SOD2；吉非替尼耐药】.

Topics: Antineoplastic Agents; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cigarette Smoking; Drug Resistance, Neoplasm; ErbB Receptors; Gefitinib; Humans; Lung Neoplasms; Reactive Oxygen Species; Sincalide; Sirtuin 3

MiR-19b-3p has been reported in several types of human cancer. Nevertheless, the expression profile and biological functions of miR-19b-3p remain unclear in non-small cell lung cancer (NSCLC). The expression level of miR-19b-3p was evaluated in NSCLC tissues and cell lines using qRT-PCR. Survival analysis was performed using Kaplan-Meier curves, while the prognostic significance of miR-19b-3p was analyzed using Cox regression analysis in 80 NSCLC patients. The effects of miR-19b-3p on cell proliferation and invasion capacities were analyzed using CCK-8, crystal violet, and transwell assays. Target genes of miR-19b-3p were assessed using luciferase reporter assay, qRT-PCR, Western blot and rescue experiments. MiR-19b-3p was found to be upregulated in human NSCLC tissues and cell lines. The expression of miR-19b-3p was observed to be closely associated with TNM stage and metastasis. High expression of miR-19b-3p was found to be capable of predicting poor clinical prognosis in NSCLC patients. Whilst overexpression of miR-19b-3p was demonstrated to promote the proliferation and invasion of NSCLC cells, knockdown of miR-19b-3p showed an opposite inhibitory effect. Bioinformatics analysis and luciferase reporter assays confirmed that HOXA9 is a direct target of miR-19b-3p. Functional assays demonstrated that NSCLC cell proliferation and invasion were promoted by miR-19b-3p via negative regulation of HOXA9. Finally, overexpression of HOXA9 was shown to partially reverse the tumor promoting effect of miR-19b-3p. This study indicates that miR-19b-3p is a crucial prognostic biomarker of NSCLC, and that targeting of the miR-19b-3p/HOXA9 axis may be a promising strategy in NSCLC therapy.

Topics: Biomarkers; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Movement; Cell Proliferation; Gene Expression Regulation, Neoplastic; Gentian Violet; Humans; Lung Neoplasms; MicroRNAs; Prognosis; Sincalide

Lung squamous cell carcinoma (LSCC) is a prevalent and progressive subtype of lung cancer. This study aimed to substantiate the regulatory effect of the PAK2/SOX2/DEK axis on the LSCC development. LSCC tissues (n = 83) and adjacent normal tissues were collected and SOX2 expression was determined by qRT-PCR and Western blotting. Correlation between SOX2 expression and the prognosis of LSCC patients was then explored utilizing Kaplan-Meier analysis. Co-immunoprecipitation and glutathione-S-transferase pull-down assays were conducted to validate the binding of SOX2 to DEK. Gain- and loss- of function assays were then performed on LSCC cells, with CCK-8 and Transwell assays applied to detect the malignant behaviors of cells. A mouse xenograft model of LSCC was further established for in vivo validation. The expression levels of SOX2, PAK2 and DEK were up-regulated in LSCC tissues and cells. SOX2 overexpression was correlated with poor prognosis of LSCC patients. Knockdown of SOX2 weakened the viability and the migratory and invasive potential of LSCC cells. Further, PAK2 directly interacted with SOX2. PAK2 overexpression accelerated the malignant phenotypes of LSCC cells through interplay with SOX2. Moreover, SOX2 activated the expression of DEK, and silencing DEK attenuated the malignant behaviors of LSCC cells. In conclusion, PAK2 could bind to the transcription factor SOX2 and thus activate the expression of DEK, thereby driving the malignant phenotypes of LSCC cells both in vivo and in vitro.

Topics: Animals; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Chromosomal Proteins, Non-Histone; Gene Expression Regulation, Neoplastic; Glutathione; Humans; Laryngeal Neoplasms; Lung; Lung Neoplasms; Mice; MicroRNAs; Oncogene Proteins; p21-Activated Kinases; Poly-ADP-Ribose Binding Proteins; Sincalide; SOXB1 Transcription Factors

Circular RNAs (circRNAs) are crucial for the pathogenesis of nonsmall lung cancer (NSCLC). Here, we set out to unravel the precise function of circRNA CD226 (circCD226) in NSCLC pathogenesis. The exosomes from serum specimens were observed by transmission electron microscopy. CircCD226, miR-1224-3p and high mobility group AT-hook 2 (HMGA2) were quantified by qRT-PCR, western blot and immunohistochemistry. Actinomycin D and Ribonuclease (RNase) R treatments and subcellular localization assay were used for circCD226 characterization. Cell viability, proliferation, migration, invasion and sphere formation abilities were gauged by CCK-8, EDU, wound-healing, transwell and sphere formation assays, respectively. Directed relationships among circCD226, miR-1224-3p and HMGA2 were examined by RNA pull-down, dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. The abundance of circCD226 was elevated in serum exosomes, tissues and cells of NSCLC. NSCLC serum exosomes enhanced NSCLC cell proliferation, migration, invasion and stemness. Loss of circCD226 impeded cell proliferation, migration, invasion and stemness in vitro , as well as tumor growth in vivo . Mechanistically, circCD226 sponged miR-1224-3p, and miR-1224-3p targeted HMGA2. CircCD226 involved the posttranscriptional regulation of HMGA2 through miR-1224-3p. Moreover, the miR-1224-3p/HMGA2 axis was identified as a functionally downstream effector of circCD226 in regulating NSCLC cell behaviors. Our study identifies circCD226 as a potential driver in NSCLC development depending on the regulation of miR-1224-3p/HMGA2 axis.

Topics: Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Movement; Cell Proliferation; Dactinomycin; Humans; Lung Neoplasms; MicroRNAs; Ribonucleases; RNA, Circular; Sincalide

Although our previous genome-wide association study (GWAS) has identified chromosome 2q33.1 as a susceptibility locus for non-small cell lung cancer (NSCLC), the causal variants remain unclear. The aims of this study were to identify the causal variants in 2q33.1 and to explore their biological functions in NSCLC.. CCK-8, colony formation, EdU incorporation, Transwell, and quantitative real-time polymerase chain reaction assays were applied to examine variant function. The tumor xenograft model was used to examine variant function. The missense variant rs3769823 (A > G), which caused the substitution of lysine with arginine at amino acid 14 in caspase-8 (caspase-8K14R), was identified as a potential causal candidate in 2q33.1. Compared with the wild type caspase-8 (caspase-8WT) group, the caspase-8K14R group had higher expression of caspase-8 and cleaved caspase-8. Caspase-8K14R inhibited the proliferation and metastasis of human lung cancer cell lines. These results suggested that rs3769823 is the causal variant in chromosome 2q33.1 and is involved in an apoptosis pathway, leading to a decreased risk of NSCLC.

Topics: Apoptosis; Arginine; Carcinoma, Non-Small-Cell Lung; Caspase 8; Caspases; Genome-Wide Association Study; Humans; Ligands; Lung Neoplasms; Lysine; Sincalide; TNF-Related Apoptosis-Inducing Ligand

Recent literature have indicated that the malignancy of cancer cells is modulated by hsa_circ_0000423 (named circPPP1R12A) through the way of translating protein. Herein, we investigated the role and latent mechanisms of circPPP1R12A in Non-Small Cell Lung Cancer (NSCLC).. CircPPP1R12A expression was measured by qRT-PCR. The malignancy of NSCLC was determined by CCK-8, TUNEL assay, Wound healing, Transwell and Western blotting assays. The underlying mechanisms of circPPP1R12A were confirmed by Western blotting and qRT-PCR assays.. CircPPP1R12A expression in NSCLC tissues was higher than that of neighboring normal tissues. CircPPP1R12A showed an upregulated expression in NSCLC cells. Upregulation of circPPP1R12A could promote the cell viability of NSCLC cells and reduce the apoptosis of NSCLC cells, while it could not promote cell invasion and migration. The reduction of cell viability and apoptosis was discovered in NSCLC cells with the silencing of circPPP1R12A, but circPPP1R12A knockdown does not inhibit cell invasion and migration. There was something interesting that circPPP1R12A encoding protein circPPP1R12A-73aa was found in NSCLC cells. Mutations in circPPP1R12a-73AA might disrupt the function of circPPP1ra-73AA in A549 and H1299 cells. Next, we found that circPPP1R12A caused the increased growth of NSCLC cells by activating AKT signaling pathway.. In summary, our study proved that NSCLC cell proliferation was promoted by circPPP1R12A-73aa translated from circPPP1R12A through the AKT pathway, which could throw some light on the understanding of the mechanism of NSCLC.

Topics: A549 Cells; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Movement; Cell Proliferation; Gene Expression Regulation, Neoplastic; Humans; Lung Neoplasms; MicroRNAs; Proto-Oncogene Proteins c-akt; Signal Transduction; Sincalide

This study aimed to identify the molecular mechanism underlying NEAT1 regulation of non-small-cell lung cancer (NSCLC) autophagy and apoptosis. The expression levels of NEAT1, miR-128-3p, and ADAM28 in NSCLC tissues and cells were examined using qRT-PCR. The relationships between NEAT1, miR-128-3p, and ADAM28 expression levels and prognosis of NSCLC patients were investigated using the Kaplan-Meier analysis method. The interactions between NEAT1, miR-128-3p, and ADAM28 were confirmed by dual-luciferase reporter assay or FISH assay. Cell proliferation and apoptosis were detected by CCK-8 and flow cytometry assays, respectively. Apoptosis and autophagy-related proteins Bax, cleaved caspase-3, cleaved caspase-9, Bcl-2, LC3, Beclin-1, and p62 were analyzed by western blotting. Finally, an in vivo NSCLC mouse xenograft model was established to confirm the in vitro data. We showed NEAT1 and ADAM28 expression levels were upregulated, while the miR-128-3p level was downregulated in NSCLC tissues and cells. NSCLC patients with high NEAT1 expression levels, low miR-128-3p levels, or high ADAM28 levels had significantly reduced overall survival times. Silencing of NEAT1 inhibited NSCLC cell autophagy and promoted apoptosis by sponging miR-128-3p. MiR-128-3p directly targeted ADAM28 and suppressed ADAM28 expression, which led to deactivation of the JAK2/STAT3 signaling pathway. Furthermore, ADAM28 overexpression attenuated miR-128-3p overexpression-induced apoptosis and autophagy inhibition in NSCLC by increasing the phosphorylation of JAK2 and STAT3. NEAT1 depletion or miR-128-3p overexpression inhibited autophagy and promoted apoptosis in vivo by suppressing ADAM28. In other word, silencing NEAT1 inhibited autophagy and then promoted NSCLC cell apoptosis by deactivating the JAK2/STAT3 signaling pathway through regulation of miR-128-3p/ADAM28 axis.

Topics: ADAM Proteins; Animals; Apoptosis; Autophagy; bcl-2-Associated X Protein; Beclin-1; Carcinoma, Non-Small-Cell Lung; Caspase 3; Caspase 9; Cell Line, Tumor; Cell Proliferation; Humans; Lung Neoplasms; Mice; MicroRNAs; Proto-Oncogene Proteins c-bcl-2; RNA, Long Noncoding; Sincalide

Non-small cell lung carcinoma (NSCLC) is a subtype of lung cancer with unfavorable outcome. Autophagy, a mechanism responsible for cellular component degradation, has been recorded to play either a positive or negative regulatory role in apoptosis. Tectonin Beta-Propeller Repeat Containing 1 (TECPR1) is recognized relevant to autophagy. This study aimed to investigate the molecular mechanisms through which TECPR1 regulates NSCLC cell apoptosis.. Analysis of TECPR1 expression in the subcategories of NSCLC was conducted using GEPIA. Survival analysis for NSCLC patients was performed with Kaplan-Meier's plotter. The interaction between ATG5 and TECPR1 was predicted by STRING and validated through co-immunoprecipitation. NSCLC cells were transfected with short hairpin RNA against ATG5 and/or ATG5/TECPR1 overexpression plasmids, followed by viability and apoptosis assay using CCK-8 and flow cytometry. Expressions of TECPR1, ATG5, LC3-II/LC3-I, P62, B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) in NSCLC cells with or without transfection were assessed by qRT-PCR and/or Western blot.. TECPR1 was low-expressed in LUAD and LUSC samples as well as NSCLC cells. Higher TECPR1 expression was associated with better outcomes. TECPR1 overexpression and ATG5 overexpression both decreased viability, promoted apoptosis, upregulated Bax and LC3-II/LC3-I, and downregulated P62 and Bcl-2. TECPR1 could form a complex with ATG5 in NSCLC cells. ATG5 was upregulated by TECPR1 overexpression and could positively modulate TECPR1 expression. ATG5 knockdown induced effect oppositely to TECPR1 overexpression, and this effect reversed the TECPR1 overexpression-induced effect and vice versa.. TECPR1 induces NSCLC cell apoptosis via ATG5 upregulation-induced autophagy promotion.

Topics: Apoptosis; Autophagy; Autophagy-Related Protein 5; bcl-2-Associated X Protein; Carcinoma, Non-Small-Cell Lung; Humans; Lung Neoplasms; Membrane Proteins; RNA, Small Interfering; Sincalide; Up-Regulation

Non-small cell lung cancer (NSCLC) results in high mortality and has gained increasing attention. C-Phycocyanin (C-PC) has been identified as a potential therapeutic inhibitor for NSCLC, but its underlying mechanism remains obscure. The gene expression of the long noncoding RNA neighbour of BRCAI RNA 2 (NBR2) in NSCLC cells was evaluated by quantitative reverse transcription-PCR. The cell capacity for proliferation and migration was examined by EdU and wound-healing assays. Furthermore, the viability and apoptosis of cells was measured with CCK-8 and annexin V/PI, respectively. Next, the protein level of activation of adenosine monophosphate- activated protein kinase and the rapamycin kinase (mTOR) signalling pathway-associated molecules was evaluated by western blotting. H292 cells were pre-treated with C-PC or transfected with plasmids encoding NBR2 or the shNBR2 plasmid, to over-express or knock down NBR2 expression, respectively. NBR2 expression was robustly down-regulated in NSCLC cell lines compared with a normal cell line (BEAS-2B). NBR2 over-expression inhibited migration and promoted apoptosis of H292 cells. Treatment of H292 cells with C-PC enhanced NBR2 levels in a dose- and time-dependent manner. Downregulation of NBR2 in H292 cells inhibited the activity of C-PC on cell proliferation, viability and clone formation. Further mechanistic investigation showed that the down-regulation of NBR2 abolished the modulatory effects of C-PC on the AMPK/mTOR signalling pathway. In conclusion, C-PC inhibits H292 cell growth by enhancing the NBR2/AMPK signalling pathway.

Topics: Adenosine Monophosphate; AMP-Activated Protein Kinases; Annexin A5; Apoptosis; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Movement; Cell Proliferation; Humans; Lung Neoplasms; Phycocyanin; RNA, Long Noncoding; Sincalide; Sirolimus; TOR Serine-Threonine Kinases

Cancer-associated fibroblasts (CAFs) within the tumor microenvironment are key players in tumorigenesis and tumor development. Nevertheless, the regulatory mechanisms of CAFs on lung squamous cell carcinoma- (LUSC-) associated remain poorly elucidated.. The microarray dataset GSE22874, containing 30 specimens of primary culture of normal fibroblasts (NFs) and 8 specimens of cancer-associated fibroblasts (CAFs) samples derived from LUSC, was retrieved from the Gene Expression Omnibus (GEO) database and then calculated by using the R language (limma package) to identify differentially expressed genes (DEGs). CAF-conditioned medium (CAF-CM) was collected and used to culture LUSC cells, followed by assessment of cell proliferation, apoptosis, and oxidative stress levels by using CCK-8, annexin V-FITC/PI double staining and ELISA assays. Subsequently, COL10A1 was knocked down in CAFs to assess the role of COL10A1 in CAF regulation of LUSC behavior. Bioinformatics online analysis and MeRIP were applied to predict and test the m. Elevated expression of COL10A1 was found in LUSC-derived CAFs by GSE22874 dataset analysis. We discovered that COL10A1 and METTL3 was expressed in both LUSC cells and matched CAFs, while COL10A1 expression was prominently higher in CAFs than in LUSC cells. CAF-CM memorably encouraged LUSC cell proliferation and suppressed apoptosis-induced oxidative stress, which was reversed by interfering with COL10A1 expression in CAFs, suggesting that COL10A1 might be secreted by CAFs into the culture medium to exert its effects inside LUSC cells. Global m. COL10A1 secreted by CAFs could facilitate LUSC cell proliferation and repress apoptosis-induced oxidative stress, and the mechanism was due to elevated expression mediated by METTL3 promoting its mRNA m

Topics: Animals; Apoptosis; Cancer-Associated Fibroblasts; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Collagen Type X; Culture Media, Conditioned; Fibroblasts; Lung; Lung Neoplasms; Methylation; Methyltransferases; Mice; Mice, Nude; Oxidative Stress; RNA, Messenger; Sincalide

Increasing evidence indicated that miR-2682-5p acted as a tumor suppressor in various cancers. The current study aimed to investigate the biological function of exosomal miR-2682-5p in non-small cell lung cancer (NSCLC).. The expression of miR-2682-5p in NSCLC tissues and adjacent non-tumor tissues, NSCLC cell lines and human embryonic lung fibroblast, as well as serum and serum exosomes of NSCLC patients and healthy donors was detected by RT-qPCR. The effects of miR-2682-5p on the viability, migration, and apoptosis of NSCLC cells were detected by CCK-8, Transwell, and flow cytometry assays. Dual-luciferase reporter gene and RNA immunoprecipitation assays were used to evalutate the relationship between miR-2682-5p and HDAC1.. Low expressed miR-2682-5p was found in tumor tissues, cell lines, serum, and serum exosomes of NSCLC patients. MiR-2682-5p overexpression suppressed NSCLC cell viability and migration and promoted apoptosis, while miR-2682-5p knockdown showed the opposite results. Furthermore, exosomes from healthy donor serum inhibited NSCLC cell viability and migration and promoted apoptosis. Dual-luciferase reporter gene and RNA immunoprecipitation assays verified that HDAC1 was a target of miR-2682-5p. HDAC1 overexpression abolished the effects of miR-2682-5p mimic on NSCLC cell viability, migration, and apoptosis. Chromatin immunoprecipitation assay indicated that HDAC1 bound to the promoter region of ADH1A. Upregulation of ADH1A counteracted the effects of HDAC1 overexpression on NSCLC cell viability, migration, and apoptosis.. Taken together, exosomal miR-2682-5p inhibited NSCLC cell viability and migration and promoted apoptosis by the HDAC1/ADH1A axis, and this result might provide a novel therapeutic target for NSCLC.

Topics: Alcohol Dehydrogenase; Carcinoma, Non-Small-Cell Lung; Cell Line; Cell Survival; Down-Regulation; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Gene Silencing; Histone Deacetylase 1; Humans; Lung Neoplasms; MicroRNAs; Sincalide; Up-Regulation

Ultrasound-targeted microbubble destruction (UTMD) is a novel adjuvant tumor therapeutic method by enhancing exogenous gene transfection to target tissues. This study aims to investigate the role of microRNA-492 (miR-492) in non-small cell lung cancer (NSCLC) and further analyze the effects of UTMD-mediated miR-492 inhibitor on tumorigenesis.. The expression of miR-492 was detected by qRT-PCR. Co-transfection of microbubbles and miR-492 inhibitor with Lipofectamine 3000 was performed to achieve UTMD-mediated miR-492 inhibition in NSCLC cells. CCK-8 and Transwell assay were used to determine NSCLC cell proliferation, and the migration and invasion.. High expression of miR-492 was associated with poor prognosis in NSCLC patients. miR-492 inhibitor suppressed tumor cell proliferation, migration and invasion, and UTMD not only increased the transfection efficiency of miR-492 inhibitor, but also enhance the inhibitory effects on cell biological behaviors.. The results showed that the expression level of miR-492 was up-regulated in NSCLC tissue samples and cells. Silencing of miR-492 inhibited NSCLC cell proliferation, migration and invasion, and UTMD-mediated miR-492 inhibitor could promote more significant inhibition, which indicated that UTMD-mediated miR-492 inhibitor might provide a novel strategy for the treatment of NSCLC.KEY MESSAGESmiR-492 inhibitor inhibited cell proliferation, migration and invasion.UTMD-mediated miR-492 inhibitor can promote more significant inhibition.UTMD-mediated miR-492 inhibitor provide a new strategy for NSCLC.

Topics: Aged; Aged, 80 and over; Carcinoma, Non-Small-Cell Lung; Cell Proliferation; Cell Transformation, Neoplastic; Female; Gene Expression; Humans; Lipids; Lung Neoplasms; Male; Microbubbles; MicroRNAs; Middle Aged; Real-Time Polymerase Chain Reaction; Sincalide; Transfection; Treatment Outcome; Ultrasonics; Ultrasonography

Treatment with cisplatin (DDP) is one of the standard therapies used to treat non-small-cell lung cancer (NSCLC) and fundamentally causes resistance in cancer cells, which eventually poses as an obstacle to the efficacy of chemotherapy in NSCLC. Efforts are on all over the world to explore a sensitizer of NSCLC to DDP. Here, we studied the effect of IL-7 on the resistance of NSCLC to chemotherapy. We observed that IL-7 treatment significantly enhanced DDP-induced effects in A549 and A549/DDP cells (DDP-resistant cells), including decreased cell viability and proliferation, as well as increased cell apoptosis and S arrest, indicating that IL-7 treatment resensitized DDP-resistant NSCLC cells to DDP. Subsequently, IL-7 enhanced the sensitivity of PI3K/AKT signaling and expressions of ABCG2 to DDP. By inhibiting IL-7 signaling via IL-7R knockdown or activating PI3K/AKT signaling via PI3K activation, the resensitization to DDP by IL-7 was abrogated, and the expression levels of ABCG2, p-PI3K, and p-AKT were found to be significantly higher. In vivo results also confirmed that IL-7 only in combination with DDP could remarkably induce tumor regression with reduced levels of ABCG2 in tumorous tissues. These findings indicate that IL-7, apart from its adjuvant effect, could overcome multidrug resistance of DDP to restore its chemotherapy sensitivity.

Topics: A549 Cells; Animals; Apoptosis; ATP Binding Cassette Transporter, Subfamily G, Member 2; Blotting, Western; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cisplatin; Female; Fluorescent Antibody Technique; Gene Expression Regulation, Neoplastic; Humans; Interleukin-7; Mice, Inbred BALB C; Signal Transduction; Sincalide

Mutant EGFR Non-small cell lung cancer has benefit from gefitinib, but it has limited effect for wild-type EGFR tumors. Shikonin, a natural naphthoquinone isolated from a traditional Chinese medicine, the plant Lithospermum erythrorhizon (zicao), not only can inhibit the tumor growth, but also overcome cancer drug resistance. Our aim is to investigate whether shikonin can enhance antitumor effect of gefitinib in EGFR wild-type lung cancer cells in vitro and in vivo.. CCK-8 was used to determine the proliferation of EGFR wild-type non-small cell lung cancer. Apoptosis and cell cycle were detected by flow cytometry. PKM2, STAT3, p-STAT3 and cyclinD1 were detected by Western blot. A549 tumor model was established to observe the antitumor effect of shikonin combination with gefitinib in vivo.. The results showed that combination of shikonin with gefitinib exhibited synergistic antitumor effect in vitro and in vivo. Its potential molecular mechanisms may be associated with inhibition of PKM2/STAT3/cyclinD1.. These results provide a promising therapeutic approach for the treatment of wild-type EGFR non-small cell lung cancer.

Topics: A549 Cells; Animals; Antineoplastic Agents; Antineoplastic Agents, Phytogenic; Carcinoma, Non-Small-Cell Lung; Carrier Proteins; Cell Line, Tumor; Cell Survival; Cyclin D1; Drug Synergism; ErbB Receptors; Gefitinib; Humans; Immunohistochemistry; Lung Neoplasms; Membrane Proteins; Mice; Mice, Nude; Naphthoquinones; Quinazolines; Signal Transduction; Sincalide; STAT3 Transcription Factor; Thyroid Hormone-Binding Proteins; Thyroid Hormones