NCI-H460 Human Lung Cancer Cells Research Articles

Activation of PI3K/Akt and ERK signaling pathways antagonized sinomenine-induced lung cancer cell apoptosis

Sinomenine (SIN) is a bioactive component derived from a Chinese medicinal plant. Our previous studies demonstrated that SIN has cytotoxic effects on human lung cancer cells. However, the antitumor molecular mechanisms of SIN have yet to be elucidated in detail. In the present study, we further explored the effects of SIN on NCI-H460 human lung cancer cell viability and apoptosis and investigated the regulation and function of PI3K/Akt and ERK signaling pathways during SIN-induced apoptosis in various lung cancer cell lines. NCI-H460 cells were incubated with 200 µg/ml SIN for the indicated times (0, 24, 48 or 72 h). Cell viability was assessed by MTT assay. Akt, p-Akt, ERK1/2 and p-ERK1/2 protein levels were detected by western blotting, respectively. Two different selective inhibitors (LY294002 for the PI3K pathway; PD98059 for the MEK/ERK pathway) were used to characterize the relative roles of PI3K/Akt and ERK in SIN-induced apoptosis. Apoptosis was determined by flow cytometry. SIN inhibited the proliferation of NCI-H460 cells in a time-dependent manner, which was accompanied with significant activation of pAkt and pERK. LY294002 and PD98059 both significantly increased SIN-induced apoptosis in NCI-H460, NCI-H226 and NCI-H522 cells. Our findings suggest that the activation of the PI3K/Akt and ERK signaling pathways antagonize SIN-induced lung cancer cell apoptosis and molecules that inhibit these pathways should potentiate the effects of SIN. This study represents a significant step forward in our understanding of the signal transduction pathways associated with the apoptosis elicited by SIN.

Induction of DNA damage by deguelin is mediated through reducing DNA repair genes in human non-small cell lung cancer NCI-H460 cells

It has been shown that deguelin, one of the compounds of rotenoids from flavonoid family, induced cytotoxic effects through induction of cell cycle arrest and apoptosis in many types of human cancer cell lines, but deguelin-affected DNA damage and repair gene expression (mRNA) are not clarified yet. We investigated the effects of deguelin on DNA damage and associated gene expression in human lung cancer NCI-H460 cells in vitro. DNA damage was assayed by using the comet assay and DNA gel electrophoresis and the results indicated that NCI-H460 cells treated with 0, 50, 250 and 500 nM deguelin led to a longer DNA migration smear based on the single cell electrophoresis and DNA fragmentation occurred based on the examination of DNA gel electrophoresis. DNA damage and repair gene expression (mRNA) were evaluated by using real-time PCR assay and the results indicated that 50 and 250 nM deguelin for a 24-h exposure in NCI-H460 cells, decreased the gene levels of breast cancer 1, early onset (BRCA1), DNA-dependent serine/threonine protein kinase (DNA-PK), O6-methylguanine-DNA methyltransferase (MGMT), p53, ataxia telangiectasia mutated (ATM) and ataxia-telangiectasia and Rad3-related (ATR) mRNA expressions. Collectively, the present study showed that deguelin caused DNA damage and inhibited DNA damage and repair gene expressions, which might be due to deguelin-inhibited cell growth in vitro.

Inhibition of cell motility and invasion by HangAmDan-B in NCI-H460 human non-small cell lung cancer cells

Correlation between inhibition of cell motility and anti-invasive activity by the water extract of HangAmDan-B (HAD-B), a crude extract of eight Korean medicinal animals and plants, in NCI-H460 human non-small cell lung cancer (NSCLC) cells was investigated. Within the concentrations that were not cytotoxic, HAD-B induced significant concentration-dependent inhibition of cell motility and invasiveness of NCI-H460 cells. Treatment with HAD-B resulted in dose-dependent inhibition of the activities of matrix metallo-proteinase (MMP)-2 and MMP-9, and this was correlated with a decrease in expression of their mRNA and proteins, and upregulation of tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 expression. Anti-invasive activity of HAD-B was also found to be associated with increased tightness of the tight junction (TJ), as demonstrated by an increase in transepithelial electrical resistance. In addition, the present results indicated that treatment with HAD-B resulted in repression of the levels of claudin family members, which are major components of TJs that play a key role in control and selectivity of para-cellular transport. Although further studies are needed, findings from the present study indicate that TJs and MMPs are critical targets of HAD-B-induced anti-invasiveness in NCI-H460 NSCLC cells.

Lipid-Soluble Ginseng Extract Induces Apoptosis and G0/G1 Cell Cycle Arrest in NCI-H460 Human Lung Cancer Cells

This study was performed to elucidate the anticancer mechanism of a lipid-soluble ginseng extract (LSGE) by analyzing induction of apoptosis and arrest of cell cycle progression using the NCI-H460 human lung cancer cell line. Proliferation of NCI-H460 cells was potently inhibited by LSGE in a dose-dependent manner. The cell cycle arrest at the G0/G1 phase in NCI-H460 cells was induced by LSGE. The percentage of G0/G1 phase cells significantly increased, while that of S phase cells decreased after treatment with LSGE. The expression levels of cyclin-dependent kinase2 (CDK2), CDK4, CDK6, cyclin D3 and cyclin E related to G0/G1 cells progression were also altered by LSGE. In addition, LSGE-induced cell death occurred through apoptosis, which was accompanied by increasing the activity of caspases including caspase-8, caspase-9 and caspase-3. Consistent with enhancement of caspase activity, LSGE increased protein levels of cleaved caspase-3, caspase-8, caspase-9, and poly-ADP-ribose polymerase (PARP). These apoptotic effects of LSGE were inhibited by the pan-caspase inhibitor Z-VAD-fmk. These findings indicate that LSGE inhibits NCI-H460 human lung cancer cell growth by cell cycle arrest at the G0/G1 phase and induction of caspase-mediated apoptosis.

α-Tomatine Suppresses Invasion and Migration of Human Non-Small Cell Lung Cancer NCI-H460 Cells Through Inactivating FAK/PI3K/Akt Signaling Pathway and Reducing Binding Activity of NF-κB

α-Tomatine, isolated from Lycopersicon esculentum Linn., is a naturally occurring steroidal glycoalkaloid in immature green tomatoes. Some reports demonstrated that α-tomatine had various anticarcinogenic properties. The purpose of this study is to investigate the anti-metastatic effect of α-tomatine in NCI-H460 human non-small cell lung cancer cells. First, the results showed that α-tomatine significantly suppressed the abilities of the adhesion, invasion, and migration of NCI-H460 cells under non-cytotoxic concentrations. Molecular data also showed α-tomatine could inhibit the activation of focal adhesion kinase (FAK) and phosphatidylinositol 3-kinase (PI3K)/Akt signal involve in the downregulation the enzyme activities, protein and messenger RNA levels of matrix metalloproteinase-7 (MMP-7). Next, α-tomatine also strongly inhibited the degradation of inhibitor of kappaBα (IκBα) and the nuclear levels of nuclear factor kappa B (NF-κB). Also, a dose-dependent inhibition on the binding ability of NF-κB by α-tomatine treatment was further observed. Furthermore, α-tomatine significantly decreased the levels of phospho-Akt and MMP-7 in Akt1-cDNA-transfected cells concomitantly with a marked reduction on cell invasion and migration. Presented results indicated α-tomatine might be further application for treating cancer metastasis.

The G protein-coupled receptor T-cell death-associated gene 8 (TDAG8) facilitates tumor development by serving as an extracellular pH sensor

Tumors often are associated with a low extracellular pH, which induces a variety of cellular events. However, the mechanisms by which tumor cells recognize and react to the acidic environment have not been fully elucidated. T-cell death-associated gene 8 (TDAG8) is an extracellular pH-sensing G protein-coupled receptor that is overexpressed in various tumors and tumor cell lines. In this report, we show that TDAG8 on the surface of tumor cells facilitates tumor development by sensing the acidic environment. Overexpression of TDAG8 in mouse Lewis lung carcinoma (LLC) cells enhanced tumor development in animal models and rendered LLC cells resistant to acidic culture conditions by increasing activation of protein kinase A and extracellular signal-regulated kinase in vitro. Moreover, shRNA-mediated knockdown of endogenous TDAG8 in NCI-H460 human non-small cell lung cancer cells reduced cell survival in an acidic environment in vitro as well as tumor development in vivo. Microarray analyses of tumor-containing lung tissues of mice injected with TDAG8-expressing LLC cells revealed up-regulation of genes related to cell growth and glycolysis. These results support the hypothesis that TDAG8 enhances tumor development by promoting adaptation to the acidic environment to enhance cell survival/proliferation. TDAG8 may represent a therapeutic target for arresting tumor growth.

Antitumor Effect of Skin of Venenum Bufonis in a NCI-H460 Tumor Regression Model

This experimental study was performed to investigate the antitumor effect of skin of venenum bufonis (SVB) in NCI-H460 human lung cancer cell xenografted nude mice. NCI-H460 cell lines were cultured and then xenografted into nude mice. Mice were divided into four groups: SVB (0.25 mg/kg) given orally, SVB (0.25 mg/kg) interperitoneally, SVB (0.5 mg/kg) interperitoneally, and the untreated group. Mice were raised and treated for 28 days. Body weight and tumor weight and volumes were measured daily. Absolute organ weight, microhistological observations and biochemical blood analyses were performed on the final day of the study following the sacrificing of these animals. Tumor inhibition rate, mean survival time and percent increase in life span were also calculated. Tumor size decreased in all SVB treated mice. Increasing the dose of SVB attenuated the inhibition rate seen on the 11th day of this experiment. Furthermore, tumor weight and volume in the mice treated with the highest dose of SVB were the smallest. Mice treated with high-dose intraperitoneal SVB gained weight and had significantly smaller spleens compared with untreated mice. Mean survival time and percent increase in life span in the low-dose intraperitoneal SVB treatment group were higher than those of other groups. Biochemical blood analysis revealed that phosphatase and urea nitrogen levels were decreased significantly in 0.25 mg/kg SVB orally treated mice ( p < 0.01). Blood level calcium and alanine transaminase significantly decreased with intraperitoneal SVB 0.5 mg/kg ( p < 0.05). The findings of this in vivo study suggest that SVB may have potential as a tumor growth inhibitor. Further research that overcomes the limitations of this study to determine the antitumor mechanism of SVB is still required.

Cytotoxic effect of a polymer–copper(II) complex containing 2,2′-bipyridyl ligand on human lung cancer cells

The cytotoxic activity of the polymer–copper(II) complex, [Cu(bpy)2(BPEI)]Cl2·4H2O (bpy = 2,2′-bipyridine, BPEI = branched polyethyleneimine) on NCI-H460 human lung cancer cells was determined adopting MTT assay and specific staining techniques. Many of the cultured NCI-H460 cancer cells treated with this complex suffered loss of viability and death mostly through apoptosis and/or necrosis as evidenced by the changes in the nuclear morphology and cytoplasmic features.

Anticancer effect of mountain ginseng Pharmacopuncture to the nude mouse of lung carcinoma induced by NCI-H460 human non-small cell lung cancer cells

Objectives : This study was performed to examine the anticancer effect of mountain ginseng Pharmacopuncture(MGP) to the nude mouse of lung carcinoma induced by NCI-H460 human nonsmall lung cancer cells. Methods : Human lung cancer (NCI-H460) cells were cultured and applied to evaluate anti-tumor activity in nude mice. After confirmed tumor growth in mice, MGP was treated per 0.1ml/kg dose to intraperitoneal and intravenous injection everyday for four weeks. And checked the changes in body weights, tumor volume, mean survival time and percent, increase in life span, histo-pathological findings, organ weights, and blood chemistry levels. Results : The results of in vivo study showed that MGP may have potential as growth inhibitor of solid tumor induced NCI-H460 without marked side effects. MGP inhibited dosage-dependently the growth of NCI-H460 cell-transplanted solid tumor compared with the control group. And mean survival time of MGP treated group was prolonged comparing with control group. Generally the group of intravenous injection is more effective than intraperitoneal injection. Conclusion : These results were suggested that MGP may be a useful anticancer agent for therapy of human lung cancer. And follow study need for the certain evidence.

Curcumin inhibits human lung large cell carcinoma cancer tumour growth in a murine xenograft model

Curcumin can decrease viable cells through the induction of apoptosis in human lung cancer NCI-H460 cells in vitro. However, there are no reports that curcumin can inhibit cancer cells in vivo. In this study, NCI-H460 lung tumour cells were implanted directly into nude mice and divided randomly into four groups to be treated with vehicle, curcumin (30 mg/kg of body weight), curcumin (45 mg/kg of body weight) and doxorubicin (8 mg/kg of body weight). Each agent was injected once every 4 days intraperitoneally (i.p.), with treatment starting 4 weeks after inoculation with the NCI-H460 cells. Treatment with 30 mg/kg and 45 mg/kg of curcumin or with 8 mg/kg of doxorubicin resulted in a reduction in tumour incidence, size and weight compared with the control group. The findings indicate that curcumin can inhibit tumour growth in a NCI-H460 xenograft animal model in vivo.

Cytotoxic Coumarins from the Bark of Mammea siamensis

A new geranylated coumarin, (E)-4-(1-hydroxypropyl)-5,7-dihydroxy-6-(3,7-dimethyl-2,6-octadienyl)-8-(3-methyl-1-oxobutyl)coumarin (named surangin D), was isolated from the bark of Mammea siamensis collected in Vietnam, along with four known coumarins, surangins B and C, and theraphins B and C, and seven xanthones, 1,7-dihydroxyxanthone, 7-hydroxy-1-methoxyxanthone, 1,7-dimethoxyxanthone, 1,7-dimethoxy-6-hydroxyxanthone, 1,6,7-trihydroxyxanthone, 1,3,7-trihydroxyxanthone, and 1,7-dihydroxy-3-methoxyxanthone. Their structures were determined by spectroscopic methods (mainly 1D- and 2D-NMR) and preparation of methylated derivatives. The four coumarins, surangins C and D and theraphins B and C, were tested for inhibition of cell proliferation in DLD-1 (colon cancer), MCF-7 (breast adenocarcinoma), HeLa (human cervical cancer) and NCI-H460 (human lung cancer) cell lines using the sulforhodamine B (SRB) assay. In all four cell lines, theraphin C showed the strongest activity (IC₅₀ in the range of 1.6-5.7 µM). Testing the anti-proliferative effect of the methylated derivatives showed reduced cellular effects of all derivatives, indicating that the number and position of free hydroxyl groups were very important for the anti-proliferative effect.

Effects of sinomenine on proliferation and apoptosis in human lung cancer cell line NCI-H460 in vitro

Sinomenine (SIN) is an herbal ingredient isolated from Sinomenium acutum Rehd. et Wils. We evaluated the effects of SIN on the growth and apoptosis of human lung cancer NCI-H460 cells. SIN inhibited the proliferation of the cells in a dose- and time-dependent manner. Apoptosis was confirmed by flow cytometry, the TUNEL assay and transmission electron microscopy. SIN-induced apoptosis was accompanied by the collapse of mitochondrial membrane potential, the release of cytochrome C and the activation of caspase-9 and -3. SIN also increased Bax and decreased Bcl-2 protein levels in the NCI-H460 cells. Data analysis suggested that SIN exerted a significant apoptotic effect on NCI-H460 cells through the mitochondrial pathway. SIN is therefore a promising chemopreventive candidate for lung carcinoma.

난담반의 인체폐암세포주 NCI-H460에 대한 세포자살유도 효능

Background : Anticancer effects of herbal medicine have been reported in various types of cancer, but the systematic approaches to explain molecular mechanism(s) are not established yet. Objective : The purpose of this study is to investigate the apoptotic cell death by Egg White combined Chalcanthite in NCI-H460 human lung cancer cells. Methods : Inhibitory effects were estimated by the MTT-assay. Cancer cells were stained with DAPI and showed condensed and fragmented nuclei. The expression of cleaved caspase-3, bcl-2, and bax was detected by western blotting. To establish a basis of understanding for anti-cancer mechanism, whole proteins have been obtained from NCI-H460 harvested at 24 hrs after the treatment of Egg White combined Chalcanthite, protein expression has been profiled by 2DE-based proteomic approach. Results : NCI-H460 human lung cancer cells were treated by three samples of IS3, IS4 and IS5. IS4 inhibited most effectively the growth of NCI-H460 human lung cancer cells. The expression of cleaved caspase-3 increased in IS4 in a concentration-dependent manner. Various changes of the protein expression have been monitored, and most frequent dysregulation was found in Vimentin, Lamin-A/C. Conclusion : Egg White combined-Chacanthite inhibited the growth of NCI-H460 human lung cancer cells by inducing the apoptotic cell death via caspase-3 activation. Based upon the present findings, the further study will focus on monitoring various cancer survival factors after artificial regulation of the proteins identified, and it would be the basis for the understanding of the Chacabthite anticancer effect(s) at the molecular level.

The transcription factor ZBP‐89 suppresses p16 expression through a histone modification mechanism to affect cell senescence

The transcription factor ZBP-89 has been implicated in the induction of growth arrest and apoptosis. In this article, we demonstrate that ZBP-89 was able to restrain senescence in NCI-H460 human lung cancer cells, through epigenetically regulating p(16INK4a) expression. Specifically, our results indicate that knockdown of ZBP-89 by RNA interference stimulated cellular senescence in NCI-H460 cells, as judged by the senescence-associated beta-galactosidase activity assay and senescence-associated heterochromatin foci assay, and this process could be reversed by RNA interference-mediated p16(INK4a) silencing. We also show that histone deacetylase (HDAC) 3 and HDAC4 inhibited p16(INK4a) promoter activity in a dose-dependent manner. Furthermore, chromatin immunoprecipitation assays verified that HDAC3 was recruited to the p16(INK4a) promoter by ZBP-89 through an epigenetic mechanism involving histone acetylation modification. Moreover, immunofluorescence and coimmunoprecipitation assays revealed that ZBP-89 and HDAC3 formed a complex. These data suggest that ZBP-89 and HDAC3, but not HDAC4, can work coordinately to restrain cell senescence by downregulating p16(INK4a) expression through an epigenetic modification of histones.

Effect of Compound K, a Metabolite of Ginseng Saponin, Combined with γ-Ray Radiation in Human Lung Cancer Cells in Vitro and in Vivo

Pretreatment of NCI-H460 human lung cancer cells with compound K produced by intestinal bacteria enhances gamma-ray radiation-induced cell death. Increases in apoptosis induced by combined treatment are made apparent in the observation of nuclear fragmentation, loss of mitochondrial membrane potential (Deltapsi), and activation of caspase 3. Apoptosis induced by compound K and gamma-ray radiation is associated with reactive oxygen species (ROS) generation. Furthermore, compound K, in combination with gamma-ray radiation, has an enhanced effect in the regression of NCI-H460 tumor xenografts of nude mice. These results suggest that compound K has possible application for cancer therapy when used in combination with gamma-ray radiation.

Antitumor activity of cytokine-induced killer cells against human lung cancer

Lung cancer is the leading cause of cancer-related death among men and women in the world. Despite the aggressive treatment with surgery, radiation and chemotherapy, the long term survival for lung cancer patients remains low. In this study, the anti-tumor activity of cytokine-induced killer (CIK) cells against human lung cancer was evaluated in vitro and in vivo. Although CD3+CD56+ CIK cells were rare in fresh human peripheral blood mononuclear cells, they could expand more than 1000-fold on day 14 in the presence of anti-CD3 antibody plus IL-2. At an effector-target cell ratio of 30:1, CIK cells destroyed 98% of NCI-H460 human lung cancer cells, which was determined by the 51Cr-release assay. In addition, CIK cells at doses of 3 and 30 million cells per mouse inhibited 57% and 77% of NCI-H460 tumor growth in nude mouse xenograft assay, respectively. This study suggests that CD3+CD56+ CIK cells may be used as an adoptive immunotherapy for patients with lung cancer.

Saxatilin Suppresses Tumor-induced Angiogenesis by Regulating VEGF Expression in NCI-H460 Human Lung Cancer Cells

Tumor growth and metastasis are dependent on angiogenesis, and endothelial cell invasion and migration are apparent means of regulating tumor progression. We report here that saxatilin, a snake venom-derived disintegrin, suppresses the angiogenesis-inducing properties of NCI-H460 human lung cancer cells. Culture supernatants of NCI-H460 cells are able to induce human umbilical vascular endothelial cell (HUVEC) invasion and tube formation. However, treatment of the cancer cells with saxatilin resulted in reduced angiogenic activity of the culture supernatant. This suppressed angiogenic property was found to be associated with the level of vascular endothelial growth factor (VEGF) in the culture supernatant. Further experimental evidence indicated that saxatilin inhibits VEGF production in NCI-H460 cells by affecting hypoxia induced factor-1 alpha (HIF-1 alpha) expression via the Akt pathway.

Neurotoxic meroditerpenoids from the tropical marine brown alga Stypopodium flabelliforme.

Brine shrimp toxicity and TLC analysis guided the isolation of five new and biologically active meroditerpenoids [2beta,3alpha-epitaondiol (1), flabellinol (2), flabellinone (3), stypotriolaldehyde (4), and stypohydroperoxide (5)] along with five known compounds from the marine brown alga Stypopodium flabelliforme collected in Papua New Guinea. The planar structures of compounds 1-5 were determined by extensive spectroscopic analysis (1D and 2D NMR, LRMS, HRMS, IR, and UV), while relative configuration was determined by 1D and 2D NOE experiments. X-ray crystallography confirmed the relative configuration of 2beta,3alpha-epitaondiol (1), and the modified Mosher's ester method was used to establish its absolute configuration. All of the new metabolites were moderately toxic to murine neuro-2a cells (LC50 2-25 microM), and three [2beta,3alpha-epitaondiol (1), flabellinol (2), and flabellinone (3)] possessed potent sodium channel blocking activity. Stypotriolaldehyde (4) had a biphasic effect on the concentration of intracellular Ca2+ in rat cerebellar granule neurons (CGN). The previously known compound, stypoldione (6), also modulated intracellular calcium concentration and was cytotoxic in CGN. Metabolites 2beta,3alpha-epitaondiol (1), flabellinol (2), and flabellinone (3) displayed moderate cytotoxicity to the NCI-H460 human lung cancer cell line.

In vitro Cytotoxicity of Extracts from Brazilian Asteraceae

Aqueous and organic extracts of Asteraceae (Compositae) collected from the State of Rio Grande do Sul, Brazil, have been tested in vitro for cytotoxic activity against human solid tumour cell lines. Twenty-five species, 125 extracts in total, were screened against HT29 human colon adenocarcinoma cells and NCI-H460 human non-small cell lung cancer cells. Twenty-five extracts from 11 species demonstrated cytotoxicity at 100µg/ml against one or both of the cell lines tested. Further analysis was performed on the active extracts using three cell lines HT29, NCI-H460, and U373 human glioblastoma cells, to determine the IC 50 and the degree of tumour cell line selectivity. Extracts from Baccharis coridifolia, Baccharis ochracea, Eupatorium macrocephalum, Eupatorium pedunculosum and Stenachaenium riedelii all produced IC 50 values below 5µg/ml. Comparison of the IC 50 results between cell lines identified that Baccharis coridifolia, Baccharis ochracea, Eupatorium laevigatum and Pluchea sagittalis extracts produced differential sensitivity across the panel of three cell lines. These species are currently under further investigation with the ultimate objective of isolation and identification of the active principles responsible for the anti-proliferative activity.

C-erbB-2/neu overexpression enhances metastatic potential of human lung cancer cells by induction of metastasis-associated properties: Yu D, Wang S-S, Dulski KM, Tsai C-M, Nicolson GL, Hung M-C Department of Tumor Biology, Texas Univ. M. D. Anderson Can. Ctr., Box 79, 1515 Holcombe Blvd., Houston, TX 77030. Cancer Res 1994; 54:3260–3266

Previously, we and others have reported high levels of expression of the c-erbB-2/neu gene in non-small cell lung cancer cell lines and primary tumors. We have also found that expression of c-erbB-2/neu-encoded p185neu was correlated with lymph node metastasis in lung squamous cell carcinomas. To investigate the potential role of the c-erbB-2/neu gene in lung cancer metastasis systematically, we introduced the human c-erbB-2/neu gene into very low p185neu-expressing NCI-H460 human non-small cell lung cancer cells and then examined the experimental metastatic potentials among the parental NCI-H460 cells and stable transfectants with increased expression of p185neu. Compared with the parental NCI-H460 cells, the NCI-H460 transfectants overexpressing p185neu produced significantly more pulmonary and extrapulmonary metastatic tumors in nude mice. The changes in experimental metastatic potential in vivo were accompanied by increased invasiveness in vitro. In addition, important steps in the invasion and metastasis process, such as secretion of basement membrane-degradative enzymes and migration through reconstituted basement membrane (Matrigel), were also increased in the NCI-H460 transfectants overexpressing p185neu. Moreover, scanning electron microscopy revealed that the p185neu-overexpressing NCI-H460 transfectants had significantly more microvilli and membrane protrusions than the parental cells, correlating with the increased invasive properties of these cells. The results demonstrate that overexpression of p185neu can enhance the experimental metastatic potential of NCI-H460 human lung cancer cells by promoting invasion and the other steps in the metastatic cascade.