Growth Of A549 Research Articles

Mechanisms for inhibition effects of polypeptide extract from scorpion venom (PESV) on proliferation of A549 cell lines in vitro

To investigate the mechanisms for inhibition effects of PESV on proliferation of non-small cell lung cancer cell line A549. MTT was used to observe cell growth and proliferation of A549 at different concentrations of PESV. Flow cytometry (FCM) was applied to analyze cell cycle distribution. Immunocytochemistry and western blot assay was recruited to detect the expression of VEGF, HIF-1alpha, PTEN after the intervention of PESV. A549 cells may be arrested mainly in G0/G1 phase and cell proliferation was significantly inhibited (P < 0.01) after PESV intervention in a certain range of concentration. PESV can significantly reduce the expression of HIF-1alpha,VEGF and increase the expression of PTEN. PESV can block cell cycle and inhibit angiogenesis directly to inhibit cell proliferation of non-small cell lung cancer cell line A549 mainly through reducing the expression of HIF-1alpha, VEGF and increasing the expression of PTEN.

Synthesis of ferrocenyl pyrazole-containing chiral aminoethanol derivatives and their inhibition against A549 and H322 lung cancer cells

A series of novel ethyl 3-ferrocenyl-1-(2-hydroxy-3-(phenylamino)propyl)-1H-pyrazole-5-carboxylate derivatives with optical activity (4) was synthesized by microwave-assisted reaction of substituted aniline and ethyl 3-ferrocenyl-1-(oxiran-2-ylmethyl)-1H-pyrazole-5-carboxylate that was prepared from ethyl 3-ferrocenyl-1H-pyrazole-5-carboxylate and (R)- or (S)-oxiran-2-ylmethyl 4-methylbenzenesulfonate. Structures of the compounds were characterized by means of IR, 1H NMR and mass spectroscopy. Preliminary biological evaluation showed that all of the compounds could suppress the growth of A549 and H322 lung cancer cells. Among all of the tested compounds 4a, 4b and 4d were more effective and might perform their action through cell cycle arrest. Moreover, although the inhibition differences between R and S enantiomers are mostly not so significant, (R)-4b displayed more effective inhibition than (S)-4b.

Abstract 4082: Global miRNA expression changes induced by HDAC-42 and 5-azaCdr in lung cancer cells

Abstract Introduction: Epigenetic alterations are universally seen in lung and other cancers. These include DNA methylation and histone modifications, both pharmacologically targetable abnormalities by DNA methyltransferase (DNMT) inhibitors and histone deacetylase (HDAC) inhibitors. Despite preclinical evidence, epigenetic modifiers have, to date, been ineffective in the treatment of most solid tumors. MicroRNAs are small non-coding RNAs that impact RNA and protein expression through imperfect base pairing in the 3′UTRs of transcribed genes. We investigated the ability of epigenetic drugs to affect the miRNA expression from four lung cancer cell lines, A549, H1299, H719, H520 (representing adeno, large cell, small cell and squamous carcinomas respectively). Materials and Methods: MTT was used to evaluate cell growth, while Western immunoblot and immunofluorescence was used to detect histone H3 acetylation. Annexin-V by flow cytometry was used for apoptosis detection. NanoString was used for the global profiling of miRNAs and corresponding statistical methods was performed by the Biostatistical Core of OSU-CCC. Results: We evaluated the impact of 6 HDAC inhibitors on cell growth of A549 and H1299 cell lines. HDAC-42 was among the most potent drugs and inhibited growth at nM concentrations. Both HDAC-42 and 5-azaCdr increased apoptosis in all 4 cell lines. The most significant synergy between HDAC-42 and 5-azaCdr was in H719 in which apoptosis increased from 4.9 to 38.0% at 500 nM. As expected, synergistic histone H3 acetylation was also seen. Following 72 hour treatment, RNA was harvested and global miRNA expression by NanoString showed that the expression patterns of miRNAs are different among the 4 cell lines after treatment with 200 nM HDAC-42 and 3 µM 5-azaCdr. 5-azaCdr caused more miRNA difference than HDAC-42 (19/3 differential expressed miRNAs in 5-azaCdr treated and HDAC-42 treated H520 cells, 21/6 in H719, 29/9 in H1299, 9/8 in A549). We found that some miRNA changes were common across cell lines in the HDAC-42 treated group, (increased miR129-3p in HDAC-42 treated H1299, H719 and H520 and increased miR148a in HDAC-42 treated H1299, A549 and H719 cells). We also found that 5-azaCdr induced changes in miRNA expression patterns were similar between H520, H719 and H1299 (16 out of 20 miRNAs in H520, 16 out of 21 in H719, 11 out of 29 in H1299). The synergistic activity of 5-azaCdr and HDAC-42 was observed with several individual miRNAs like miR-129-3p and miR-515-5p. Conclusion: Both HDAC-42 and 5-azaCdr could regulate the expression of certain miRNAs in lung cancer cells. The effect of HDAC-42 and 5-azaCdr on four different lung cancer cell lines showed distinct miRNA expression patterns. The role of several candidate miRNAs such as miR-129-3p, miR-515-5p, miR-148a and miR-200c related to acetylation and methylation requires further study. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4082. doi:1538-7445.AM2012-4082

Abstract LB-243: Cetuximab and Artesunate synergistically inhibit the invasion and distant metastasis of non-small cell lung cancer

Abstract Cancer is a complex disease and multi step process evolving from malfunctions of molecules and their complex networks which regulate this process. The blockage of the cancer signalling network is crucial in the cancer treatment. In some cases, single drug therapy might not be comprehensive enough to inhibit the activity of secondary signals, feedback regulations and resistance to particular molecules, due to their abundant expression or activity. An understanding of these complex phenomena in cancer therapy for the benefit of patients is essential, especially since it bears the chance to establish novel concepts of combination drug therapy. In the present study, we have tried to find a compound combination to overcome resistance towards Cetuximab, which blocks ligand binding to the epidermal growth factor receptor (EGFR), in non-small cell lung cancer (NSCLC). The anti-malarial drug Artesunate (ART) has been shown to have a profound cytotoxic action against cancer cell lines, and we recently showed its anti-metastatic action in NSCLC. Initial screening experiments in NSCLC cell lines showed that Cetuximab sensitivity is positively correlated with the E-cadherin expression, whereas ART sensitivity was negatively associated with EGFR expression. A549 (high E-cadherin and EGFR, sensitive to Cetuximab alone and not sensitive to ART alone) and H1299 (no E-cadherin and medium EGFR, resistant to Cetuximab alone and sensitive to ART alone) were chosen as model cell lines for this study. Cell proliferation and colony formation assays were performed either with Cetuximab/ART alone or with the combination of both. Interestingly, Cetuximab alone was not able to reduce cell proliferation and colony formation significantly in comparison to ART. In contrast to that combination of both of the drugs able to reduce these phenomena significantly even more than ART alone (p≤0.05). Combination isobologram analysis for ART and Cetuximab showed a significantly reduced proliferation rate in both the tested cell lines. In vivo, chorioallantoic membrane (CAM) metastasis assays revealed that combinatorial treatment of the cells with both drugs significantly reduced primary tumor growth of A549 (p=0.01) and H1299 (p=0.02) cells, in contrast to the treatment by Cetuximab alone. Our findings suggest that ART is resensitizing Cetuximab resistant NSCLC cells for treatment with this antibody. Currently, our study will be completed by experiments to identified major molecules and signalling cascades mediating compound interaction and re-sensitization. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-243. doi:1538-7445.AM2012-LB-243

Cytotoxic triterpene saponins from the leaves of Aralia elata

Phytochemical investigation of the leaves of Aralia elata has led to the isolation of four new compounds, 3-O-β-d-glucopyranosyl (1→3)-β-d-glucopyranosyl (1→3)-β-d-glucopyranosyl oleanolic acid (1), 3-O-[β-d-glucopyranosyl (1→3)-β-d-glucopyranosyl (1→3)]-[β-d-glucopyranosyl (1→2)]-β-d-glucopyranosyl hederagenin 28-O-β-d-glucopyranoside (2), 3-O-{[β-d-glucopyranosyl (1→2)]-[β-d-glucopyranosyl (1→3)-β-d-glucopyranosyl (1→3)]-β-d-glucopyranosyl} oleanolic acid 28-O-β-d-glucopyranosyl ester (3) and 3-O-[β-d-glucopyranosyl (1→2)]-[β-d-glucopyranosyl (1→3)]-β-d-glucopyranosyl caulophyllogenin (4) and two known compounds, 3-O-[β-d-glucopyranosyl (1→3)-α-l-arabinopyranosyl]-echinocystic acid (5) and 3-O-α-l-arabinopyranosyl echinocystic acid (6). The structural determination was accomplished with spectroscopic analysis, in particular 13C-NMR, 2D-NMR and HR-ESI-MS techniques. Compounds 1–6 were tested for their inhibition of the growth of HL60, A549 and DU145 cancer cells. Compound 1 showed significant cytotoxic activity against HL60 and A549 cancer cells with IC50 values of 6.99μM and 7.93μM respectively. In addition, compounds 5 and 6 showed significant cytotoxic activity against HL60 cancer cells with IC50 values of 5.75μM and 7.51μM, respectively.

Cytotoxic Triterpene Saponins of Aralia elata

The genus Aralia (Araliaceae) consists of more than 30 species, mainly distributed in Asia, some located in the northern part of America. There are about 30 species grown in China [1]. Aralias elata, which is a shrub widely distributed in northeastern China, has various biological activities, such as anti-cancer, anti-inflammatory, liver-protecting, antioxidative, antiviral and antidiabetic activities [2]. The whole plant has been used as traditional folk medicines for the treatment of neurasthenia, rheumatoid arthritis, diabetes mellitus, gastrospasm, constipation and hepatitis [3]. Flavonoids and saponins [4] had been reported from this plant. We have done a good deal of chemical investigations on the buds and root bark of Aralia elata, but very little research has been done on the leaves until now. Chemical investigation of the leaves of Aralia elata has led to the isolation of five new compounds (Congmunoside L-P, 1-5), together with eleven known triterpene saponins (6–16). Structural determination was accomplished by spectroscopic analysis, particularly 13C-NMR, 2D-NMR and TOF-MS techniques. Compounds 1-16 all had been tested for their ability to inhibit the growth of HL-60, A549 and DU145 cancer cells through MTT, and compounds 6 and 8, 10, 12, 14 show significant cytotoxic activities.

Naringenin derivative diethyl (5,4′-dihydroxy flavanone-7-yl) phosphate inhibits cell growth and induces apoptosis in A549 human lung cancer cells

Anti-cancer effects of naringenin derivative diethyl (5,4′-dihydroxy flavanone-7-yl) phosphate were evaluated in human lung cancer cells. The effect of diethyl (5,4′-dihydroxy flavanone-7-yl) phosphate (dEdHF-7-p) on A549 cell viability was measured using MTS assay and cell counting. Morphological changes were detected using phase-contrast microscopy. Apoptosis was analyzed using Hoechst staining. The influence of dEdHF-7-p on cell cycle distribution was determined using propidium iodide (PI) staining, and protein expression was determined by Western blot analysis. A newly synthesized naringenin derivative dEdHF-7-p suppressed cell growth of A549 though mechanisms including inhibition of cell cycle and increased apoptosis. Apoptotic and cell cycle modulators were changed by dEdHF-7-p in A549 cells; cyclins, ppRB, and antiapoptotic factor Bcl-2 were down-regulated, whereas apoptotic factor Bax and cyclin-dependent kinase inhibitors p21 and p53 were enhanced, thereby releasing cytochrome c into the cytosol of dEdHF-7-p -treated-A549 cells. dEdHF-7-p treatment processed caspases-3/-8/-9 and cleavage of poly ADP-ribose polymerase. The dEdHF-7-p treatment enhanced Fas expression and decreased expression of cell survival factors such as PI3K and p-Akt in a dose-dependent manner. Taken together, dEdHF-7-p induces apoptosis by inhibiting the PI3K/Akt survival signaling pathway and modulating mitochondria-emanated intrinsic and Fas extrinsic pathways in A549 cells.

Prolonged sulforaphane treatment does not enhance tumorigenesis in oncogenic K-ras and xenograft mouse models of lung cancer

Background:Sulforaphane (SFN), an activator of nuclear factor erythroid-2 related factor 2 (Nrf2), is a promising chemopreventive agent which is undergoing clinical trial for several diseases. Studies have indicated that there is gain of Nrf2 function in lung cancer and other solid tumors because of mutations in the inhibitor Kelch-like ECH-associated protein 1 (Keap1). More recently, several oncogenes have been shown to activate Nrf2 signaling as the main prosurvival pathway mediating ROS detoxification, senescence evasion, and neoplastic transformation. Thus, it is important to determine if there is any risk of enhanced lung tumorigenesis associated with prolonged administration of SFN using mouse models of cancer.Materials and Methods:We evaluated the effect of prolonged SFN treatment on oncogenic K-ras (K-rasLSL-G12D)-driven lung tumorigenesis. One week post mutant-K-ras expression, mice were treated with SFN (0.5 mg, 5 d/wk) for 3 months by means of a nebulizer. Fourteen weeks after mutant K-ras expression (K-rasLSL-G12D), mice were sacrificed, and lung sections were screened for neoplastic foci. Expression of Nrf2-dependent genes was measured using real time RT-PCR. We also determined the effect of prolonged SFN treatment on the growth of preclinical xenograft models using human A549 (with mutant K-ras and Keap1 allele) and H1975 [with mutant epidermal growth factor receptor (EGFR) allele] nonsmall cell lung cancer cells.Results:Systemic SFN administration did not promote the growth of K-rasLSL-G12D-induced lung tumors and had no significant effect on the growth of A549 and H1975 established tumor xenografts in nude mice. Interestingly, localized delivery of SFN significantly attenuated the growth of A549 tumors in nude mice, suggesting an Nrf2-independent antitumorigenic activity of SFN.Conclusions:Our results demonstrate that prolonged SFN treatment does not promote lung tumorigenesis in various mouse models of lung cancer.

A new phenylpropanoid glycoside from the fruits of Illicium simonsii

Aim To study the constituents and bioactivity of Illicium simonsii. Methods The extracts of the fruits of Illicium simonsii were submitted to a combination of chromatographic materials, silica gel, ODS column chromatography and finally preparative HPLC to obtain eight compounds which were further evaluated for their cytotoxicity by MTT method. Results A new phenylpropanoid glycolside, 2, 4-dihydroxy-allylbenzene-2- O-β-D-glucopyranoside ( 1), together with seven characteristic sesquiterpene lactones, oligandrumin B ( 2), oligandrumin D ( 3), anisatin ( 4), veranisatin D ( 5), pseudomajucin ( 6), 1α-hydroxy-3-deoxy-pseudoanisatin ( 7), 8α-hydroxy-10-deoxycyclomerrillianolide ( 8) were isolated. Conclusion Compound 1 is new. Compounds 1, 2, 3, 5–8 were isolated from this plant for the first time. None of the isolates showed inhibitory effects on the growth of non-small cell lung tumor cell line A549.

Antitumor activity of motesanib alone and in combination with cisplatin or docetaxel in multiple human non–small-cell lung cancer xenograft models

BackgroundNon–small-cell lung cancer (NSCLC) is categorized into various histologic subtypes that play an important role in prognosis and treatment outcome. We investigated the antitumor activity of motesanib, a selective antagonist of vascular endothelial growth factor receptors (VEGFR) 1, 2, and 3, platelet-derived growth factor receptor, and Kit, alone and combined with chemotherapy in five human NSCLC xenograft models (A549, Calu-6, NCI-H358, NCI-H1299, and NCI-H1650) containing diverse genetic mutations.ResultsMotesanib as a single agent dose-dependently inhibited tumor xenograft growth compared with vehicle in all five of the models (P < 0.05). When combined with cisplatin, motesanib significantly inhibited the growth of Calu-6, NCI-H358, and NCI-H1650 tumor xenografts compared with either single agent alone (P < 0.05). Similarly, the combination of motesanib plus docetaxel significantly inhibited the growth of A549 and Calu-6 tumor xenografts compared with either single agent alone (P < 0.05). In NCI-H358 and NCI-H1650 xenografts, motesanib with and without cisplatin significantly decreased tumor blood vessel area (P < 0.05 vs vehicle) as assessed by anti-CD31 staining. Motesanib alone or in combination with chemotherapy had no effect on tumor cell proliferation in vitro.ConclusionsThese data demonstrate that motesanib had antitumor activity against five different human NSCLC xenograft models containing diverse genetic mutations, and that it had enhanced activity when combined with cisplatin or docetaxel. These effects appeared to be mediated primarily by antiangiogenic mechanisms.

Purification, Characterization and Antitumor Activities of a New Protein from Syngnathus acus, an Officinal Marine Fish

Discovery and development of new antitumor agents from abundant marine fish are attracting an increasing interest. In the present study, we extracted and purified a novel antitumor protein Syngnathusin from the whole body of Syngnathus acus L., a precious marine fish traditionally used for tumors. Syngnathusin was comprised of 16 kinds of amino acids, mainly acidic amino acids. Its molecular weight was 67.3 kDa and its isoelectric point was 4.57. The N-terminal amino acid sequence of Syngnathusin was determined to be Lys-Arg-Asp-Leu-Gly-Phe-Val-Asp-Glu-Ile-Ser-Ala-His-Tyr and showed no significant homology with the known proteins. Syngnathusin could significantly inhibit the growth of A549 and CCRF-CEM cells. However, the obvious proliferation inhibition against human non-tumor cell lines was not observed. Flow cytometry, morphologic assessment and comet assay revealed that Syngnathusin could induce apoptosis in A549 and CCRF-CEM cells and strongly cooperated with MTX. Syngnathusin could inhibit the growth of S180 tumor transplanted in mice. Syngnathusin may be developed as a novel, selective and effective antineoplastic agent.

Abstract C56: Characterization of hz515H7, a novel humanized anti-CXCR4 antibody: In vitro efficacy on CXCR4-associated signaling pathways and in vivo antitumor activity.

Abstract C56: Characterization of hz515H7, a novel humanized anti-CXCR4 antibody: In vitro efficacy on CXCR4-associated signaling pathways and in vivo antitumor activity.

ChemInform Abstract: Synthesis of Novel Pyrazolo[1,5‐a]pyrazin‐4(5H)‐one Derivatives and Their Inhibition Against Growth of A549 and H322 Lung Cancer Cells.

AbstractTitle compounds (III) are obtained by a microwave‐assisted one‐step procedure.

ChemInform Abstract: Microwave‐Assisted Synthesis, Crystal Structure of Pyrazolo[1,5‐a]pyrazin‐4(5H)‐ones and Their Selective Effects on Lung Cancer Cells.

AbstractA facile microwave‐assisted route to pyrazolopyrazinones (III) (12 examples) is given.

Ruthenium methylimidazole complexes induced apoptosis in lung cancer A549 cells through intrinsic mitochondrial pathway

Ruthenium(II) methylimidazole complexes, with the general formula [Ru(MeIm)4(N⌢N)]2+ (N⌢N = tip (RMC1), iip (RMC2), dppz (RMC3), dpq (RMC4); MeIm = 1-methylimidazole, tip = 2-(thiophene-2-yl)-1H-imidazo [4,5-f] [1,10]phenanthroline, iip = 2-(1H-imidazol-4-yl)-1H-imidazo [4,5-f] [1,10]phenanthroline, dppz = dipyrido[3,2-a:2′,3′-c]phenazine, dpq = pyrazino [2,3-f] [1,10]phenanthroline), were synthesized and characterized. As determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, these complexes displayed potent anti-proliferation activity against various cancer cells. RMC1 inhibited the growth of A549 (human lung adenocarcinoma) lung cells through induction of apoptotic cell death, as evidenced by the accumulation of cell population in sub-G1 phase. RMC1 also induced the depletion of mitochondrial membrane potential in A549 cells by regulating the expression of pro-survival and pro-apoptotic Bcl-2 family members. Another experiment showed that Bid protein was also activated by RMC1, which implied that RMC1 could existed two pathways crosstalk, namely, have exogenous death receptor signaling pathway. These results demonstrated that RMC1 induced cancer cell death by acting on both mitochondrial and death receptor apoptotic pathways, suggesting that RMC1 could be a candidate for further evaluation as a chemotherapeutic agent against human cancers.

Synthesis of novel pyrazolo[1,5- a ]pyrazin-4(5 H )-one derivatives and their inhibition against growth of A549 and H322 lung cancer cells

A series of substituted pyrazolo[1,5-a]pyrazin-4(5H)-one was synthesized by the reaction of ethyl 1-(2-oxo-2-phenylethyl)-3-phenyl-1H-pyrazole-5-carboxylate derivatives and 2-(2-aminoethoxy)ethanol or 2-morpholinoethanamine in the condition of microwave-assisted one-step and solvent-free in a good yield. The structures of the compounds were determined by IR, (1)H NMR and mass spectroscopy. In addition, a representative single-crystal structure was characterized by using X-ray diffraction analysis. Preliminary biological evaluation showed that the compounds could inhibit the growth of A549 and H322 cells in dosage-dependent manners.

Abstract 4115: Prediction of drug sensitivity based on correlation of molecular and genetic signatures between lung tumor cell lines and tumor tissue samples from NSCLC cancer patients

Abstract To establish transition of preclinical response profiles to the subsequent clinical treatment, we profiled pathway activation and genomic signature in tumor cell lines and human tumor tissues. This profiling has allowed us to define molecular pathways and these models may serve as an excellent platform to evaluate the efficacy of targeted therapies. We profiled pathway activation, mutation status, and clinical histology of a panel of representative lung tumor cell lines and 50 human lung adenocarcinoma samples. We cluster patients samples with corresponding tumor cell lines based on such signature and predicted treatment options for the patients based on response to selected candidate treatment of the cell lines. Subsets of patients were identified as candidates to treatment with inhibitors of EGFR, cMET, IGFR, MEK, and PI3K/mTOR. EGF pathways in EGFR mutant cell lines NCI-H1975, HCC827 and NCI-H1650 are highly activated, and those cells were sensitive to inhibitors blocking EGFR signaling. A group of patients were identified to share the similar profiles, and we predict those are candidates for EGFRi therapies. While H1973 is a cMET amplified line with cMET and EGFR activated, patients sharing this genotype may respond to cMET inhibitor treatment. IGF1R expressing A549 and NCI-H358 cells were sensitive to IGF1R inhibition, the patients sharing the similar profile may be candidates to this class of therapies. PI3K/mTOR inhibitor, BEZ 235, effectively inhibited tumor cell growth in most of the cell lines except NCI-H1993 and NCI-H358 cells; while MEK inhibitor PD-325901 was less active in blocking growth of k-RAS mutant cell lines NCI-H460, NCI-H1734, NCI-H358 and A549. Patients with a similar signature may benefit from those treatments. By combining pathway biomarker, mutation status, and clinical features, we were able to cluster clinical patient samples to the lung tumor cell lines and propose treatment options for the patients based on their signature overlap. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4115. doi:10.1158/1538-7445.AM2011-4115

Abstract 4220: Inhibition of cancer cell growth by muscadine grape seed and grape skin extracts

Abstract The muscadine grape (Vitis rotundifolia) is native to the Southeastern United States and muscadine grape seed products are marketed as dietary supplements, based upon their antioxidant properties. Extracts from muscadine grapes contain a number of antioxidants, including resveratrol and ellagic acid; however, the natural antioxidants found in grape seed extracts are predominantly procyanidins, while the grape skin extracts contain more anthocyanins. We investigated the effect of muscadine grape seed extracts (MSE) and muscadine grape skin extracts (MSKE) on the growth of human lung, colon, prostrate, skin, brain and breast cancers as well as human leukemias. Cells were incubated with increasing concentrations (from 0.5 to 50 μg/mL) of aqueous extracts of either MSE or MSKE and the total number of cells was quantified after 7 days. Both the MSE and MSKE inhibited the growth of A549 and SK-LU-1 human lung adenocarcinoma cancer cells (81.8% inhibition at the highest concentration) and HT29 and HCT116 human colon cancer cells (80.5% inhibition at the highest dose). Similarly, extracts from both muscadine grape seeds and skins inhibited the growth of LNCaP and PC3 human prostate cancer cells. The growth of U87 and U373 human glioblastoma cells and RPMI 7951 and SKMEL28 human skin cancer cells was dose-dependently reduced by treatment with the MSE or MSKE. Human leukemia cells were also incubated with increasing concentrations of the extracts; the growth of THP-1 human acute monocytic leukemia cells, HL-60 human acute promyelocytic cells, and K562 human chronic myelogenous leukemia cells was reduced by both muscadine grape extracts (average of 74.2% inhibition at the highest dose). Both the MSE and MSKE inhibited the growth of estrogen receptor-dependent ZR-75-1, HER2 over-expressing SKBR3, and triple negative MDA-MB-231 human breast cancer cells. The inhibition of growth by either extract was highest in the triple negative breast cancer cells (92.6% inhibition at the highest concentration). The reduction in human breast cancer cell growth was accompanied by a significant decrease in the phosphorylation and activation of the mitogen-activated protein kinases ERK1 and ERK2; MAP kinase activities were reduced 91% by MSE and 80% by MSKE in ZR-75-1 estrogen receptor-dependent breast cancer cells and 73% by MSE and 66% by MSKE in MDA-MB-231 triple negative breast cancer cells. There were no significant differences between the effects of the grape seed extract compared to the grape skin extract with any of the cell lines. These results demonstrate that extracts from muscadine grape seeds and muscadine grape skins inhibit the growth of human lung, colon, prostate, breast, skin, brain and leukemia cells in vitro, suggesting that further studies are warranted to investigate their potential use in the prevention or treatment of cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4220. doi:10.1158/1538-7445.AM2011-4220

Microwave-assisted synthesis, crystal structure of pyrazolo[1,5-a]pyrazin-4(5H)-ones and their selective effects on lung cancer cells

A series of novel pyrazolo[1,5-a]pyrazin-4(5H)-one derivatives with hydrophilic group was synthesized under general heating condition and microwave-assisted condition. The structures of compounds were determined by IR, 1H NMR and HRMS, moreover, representative crystal structures were characterized by using X-ray diffraction analysis. Preliminary biological evaluation showed that some compounds could inhibit the growth of A549, H322 and H1299 cells in dosage dependent manners. The compounds could inhibit growth of A549, H322 and H1299 cells in different mechanism. Compounds 3e–h inhibited growth of A549 cells by inducing a strong G1-phase arrest. Whereas these compounds inhibited growth of H1299 and H322 cells by inducing apoptosis.

Genetic alterations of tumor suppressor ING1 in human non-small cell lung cancer

The aim of this study was to investigate the function of the ING1 gene in lung carcinoma. To detect the inhibitory effect of ING1 in human lung cancer, recombinant ING1b plasmids were transfected into two lung cancer cell lines with different p53 status, A549 with wild-type p53 (wtp53) and SK-MES-1 with mutant p53. Apoptosis, cell cycle, growth rate and the expression of downstream gene p21waf1 were analyzed. In addition, the complex of p33ING1b and p53 was analyzed with coimmunoprecipitation. To detect the gene alteration and the expression of ING1, 70 cases of fresh-frozen lung carcinomas and 217 cases of formalin-fixed, paraffin-embedded specimens were examined for loss of heterozygosity (LOH) and p33ING1b protein expression by polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and immunohistochemistry using tissue microarrays, respectively. Overexpression of ING1b inhibited the cell growth of A549 and SK-MES-1, induced cell cycle arrest and apoptosis. p21waf1 was up-regulated and a complex of p33ING1b and wtp53 was found after transfection of ING1b in the wtp53-positive lung cancer cell. High LOH frequency was found in lung carcinomas (55.7%) and p33ING1b expression was lost in 115 of 217 carcinomas (53.0%). Furthermore, there was a highly significant inverse correlation between expression and LOH frequency (P<0.05). ING1 can inhibit the growth of lung cancer cell lines through the induction of cell cycle arrest and apoptosis by forming a complex with wtp53 and up-regulating p21waf1. In human lung cancer, expression of the ING1 gene was reduced or lost and high LOH frequency of ING1 microsatellites was found. The LOH of microsatellites may down-regulate p33ING1b and/or affect its function, thereby, contributing to lung cell carcinogenesis.