Human Pulmonary Carcinoma Cell Line Research Articles

Antiproliferative Effects of Phytoestrogens Formononetin and Resveratrol Against Human Pulmonary Adenocarcinoma Cells A 549

Phytoestrogens are phytoalexins, plant secondary metabolites produced in response to microbial attacks and other forms of external stress stimuli. These compounds are devoid of the steroidal structure of endogenous estrogens but could bind with estrogen receptors with great affinity as they have structural similarity with endogenous/synthetic estrogens. Phytoestrogens are reported to possess several health benefits including cardio protective, antidiabetic, antioxidant and anti osteoporotic properties. Phytoestrogens like genistein, daidzein, and equol have been reported to possess excellent anti proliferative and anticancer properties. Formononetin, is a phytoestrogen, isoflavone widely present in red clover (Trifolium pratense) and resveratrol is a phytoestrogen which is widely present in red grapes (Vitis vinifera). The cardio protective property of red wine has been attributed to the presence of resveratrol. Although formononetin and resveratrol have been investigated extensively for several pharmacological properties, reports on their anti proliferative effects especially against human pulmonary carcinoma cell lines are very less. Hence the current study aims to understand the influence of these two compounds on the growth of human pulmonary adenocarcinoma cell line A 549. MTT assay, cytomorphology analysis by phase contrast microscopy, DNA topology assay and DPPH assay were few assays which were performed to understand the antiproliferative and antioxidant effects of these compounds. Results of the study indicated the potent antiproliferative effects of these compounds against human pulmonary carcinoma cell lines implicating that these compounds could be potential therapeutic molecules with multifaceted roles as anticancer agents, potent antioxidants and anti inflammatory agents.

Aggregation Turns BODIPY Fluorophores into Photosensitizers: Reversibly Switching Intersystem Crossing On and Off for Smart Photodynamic Therapy

Aggregation Turns BODIPY Fluorophores into Photosensitizers: Reversibly Switching Intersystem Crossing On and Off for Smart Photodynamic Therapy

Synthesis, spectroscopic characterization and biological application of copper complex of 5-carbethoxy-2-thiouracil

5-carbethoxy-2-thiouracil (eitotH2) reacts with CuX (X= Cl, Br, I) halides to give the formula [CuX(eitotH)2]2 dinuclear complexes, while the formula [CuX(PPh3)2(eitotH)2] mononuclear mixed ligand complexes result when reaction is carried out in the presence of two equivalent of triphenylphosphine (PPh3). The new copper (I) complexes were studied against two tumor cell lines, A549 (human pulmonary carcinoma cell line) and HeLa (human epithelial carcinoma cell line) and one regular immortalized cell line, MRC5 (human fetal lung fibroblast). In comparison to the phosphine free ones that hindered cell proliferation only at relatively high concentration, the mixed ligand complexes with triphenylphosphine were found to be extremely cytotoxic.
 Keywords: Copper (I), 5-carbethoxy-2-thiouracil (eitotH2), Triphenylphosphine, in vitro cytotoxicity, carcinoma cell lines

CD90-Targeted Liposomes Increase the Therapeutic Efficacy of a Retinoic Acid Derivative in Pulmonary Carcinoma

Title: The therapeutic efficacy of CD90-targeted liposomes in pulmonary carcinoma. Background: In refractory lung cancer tissue, it has been reported that cell populations exist that exhibit resistance to existing therapeutics. These cells have characteristics such as high stemness and tumorigenic potential, and they are thought to contribute to relapse and metastasis. Differentiation inducer 4[(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2- naphthalenyl) carbamoyl] benzoic acid (Am80) has a growth inhibitory effect on pulmonary carcinoma cells, but it has been effective only in a high dose that causes damage to normal cells. We aimed to achieve a dose reduction by specific targeting of anaplastic pulmonary carcinoma cells and modification of antibodies on the liposome surface specific for membrane protein of undifferentiated cells because undifferentiated pulmonary cells have unique membrane proteins. CD90 is one of the undifferentiated markers expressed in pulmonary cells. Methods and Findings: In this study, we examined the usefulness evaluation of Am80 liposomes conjugated with anti-CD90 antibody using Calu-6, a CD90-positive undifferentiated human pulmonary carcinoma cell line. After the preparation of Am80 liposomes, the characterization and cell specificity, therapeutic efficacy of them were assessed. Am80 liposomes conjugated with antibody selectively combined with the surface of Calu-6 and had a significant influence on the inhibition of cell and tumor growth. The twice-weekly intratumoral administration of Am80 liposomes conjugated antibody (0.1 mg/kg as Am80) inhibited tumor growth to 3.64 ± 1.10 compared to empty liposomes (5.89 ± 1.61) in relative tumor volume at day 28. Conclusions: We revealed that the cellular uptake of Am80 liposomes conjugated with anti- CD90 antibody by anaplastic pulmonary carcinoma cells was highly effective and efficient.

Synthesis, characterization and biological activity of tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidine derivatives as epidermal growth factor receptor inhibitors

Based on the molecular docking studies, which were performed to position Erlotinib and the target compounds into the active site of the epidermal growth factor receptor(EGFR) to determine the probable binding model, a novel series of 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidine derivatives as the novel potential EGFR kinase inhibitors was designed and synthesized. The antitumor activity of all the target compounds against human pulmonary carcinoma cell line A549 has been screened. Of all the target compounds, 4-[2-(1-piperidyl)carbonylmethoxylphenthio]- 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidine(7j) demonstrated the most potent antitumor activity. Several of the target compounds exhibited moderate antitumor activity. The preliminary structure-activity relationships of some target compounds were summarized.

Copper(I) halide complexes of 5-carbethoxy-2-thiouracil: Synthesis, structure and in vitro cytotoxicity

5-Carbethoxy-2-thiouracil (eitotH2) reacts with copper(I) halides CuX (X = Cl, Br, I) to give dinuclear complexes of the formula [CuX(eitotH2)2]2 while mononuclear mixed-ligand complexes of the formula [CuX(PPh3)2(eitotH2)] result when the reactions are performed in the presence of two equivalents of triphenylphosphine (PPh3). The molecular structures of representative compounds from each of the above types of complexes, namely [CuI(eitotH2)2]2, [CuCl(PPh3)2(eitotH2)] and [CuBr(PPh3)2(eitotH2)] have been established by single-crystal X-ray diffraction. The new copper(I) complexes were evaluated for in vitro antitumor properties against two tumor cell lines, A549 (human pulmonary carcinoma cell line) and HeLa (human epithelial carcinoma cell line) and one normal immortalized cell line MRC5 (human fetal lung fibroblast). The mixed-ligand complexes possessing triphenylphosphine were found to be highly cytotoxic in contrast to the phosphine-free ones which inhibited cell proliferation only in relatively high concentrations.

Coumarin-containing aminophosphonates bridged with chiral side chain: synthesis and influence of chirality on cytotoxicity and DNA binding

A series of novel coumarin-containing α-aminophosphonates with two chiral centers were synthesized and a single-crystal structure of compound 8g (8g′, (R)-diethyl ((S)-2-(4-methyl-2-oxo-2H-chromen-7-yloxy)propanamido)(2-bromophenyl)methylphosphonate) was obtained. The in vitro antitumor activities of compound 8a–8g′ against human pulmonary carcinoma cell line (A549), human nasopharyngeal carcinoma (human KB), and human lung adenocarcinoma (MGC-803) cell lines were evaluated. Some compounds showed relatively high cytotoxicity. Compared with 8g, 8g′ exhibited an improved activity against three tumor cells, which was evidenced by the IC50 that was four- to five-fold lower than those for 8g. The influence of chirality was also observed in DNA-binding assay of 8g and 8g′. 8g′ exhibited higher binding constant (1.96 × 103 M−1) as compared to 8g (1.69 × 103 M−1).

Effects of fudosteine, a new mucoactive drug, on the increase in mucus secretion produced by secretagogues in human pulmonary mucoepidermoid carcinoma cells

The purpose of this study was to clarify the effects of fudosteine ((—)-(R)-2-amino-3-(3-hydroxypropylthio)propionic acid), a new mucoactive drug, on mucus secretion in the human pulmonary mucoepidermoid carcinoma cell line NCI-H292. NCI-H292 cells produced hyaluronidase-resistant high-molecular-weight glycoconjugates (HMWG), which were eluted in the void volume on Superose 6HR column chromatography. ATP, bradykinin, and methacholine increased the basal secretion of [14C]glucosamine-labelled HMWG in NCI-H292 cells. Fudosteine significantly suppressed the HMWG secretion induced by ATP or bradykinin, but not that induced by methacholine. Other mucoactive drugs, such as ethylcysteine, ambroxol and carboxymethylcysteine, did not affect the increase in HMWG. These results indicate that fudosteine suppresses the mucin secretion induced by ATP and bradykinin in airway mucus-producing cancer cells.

The Lethal Effects of Nicotinamide Combined With Nonfocused Ultrasound on Human Pulmonary Carcinoma Cell Line A549 in Suspension

Objective: To explore the lethal effects of nicotinamide combined with nonfocused ultrasound (NFU) on human pulmonary carcinoma A549 cell live in suspension and provide new evidence for killing the cancer cells in hydrothorax and hydroperitonia or malignant cells infiltrating in tissues. Patients and Methods: Six groups were set in our experiment: blank control group, NA group, NFU group in 0 C, NFU group in 37 C, NA-combined NFU group in 0 C, and NA-combined NFU group in 37 C. Instant and 24-hour survival rates of A549 cells treated by different methods were determined based on the trypan blue exclusive assay, growth rates were detected by MTT after 24 hours, capability of cell colony formation was detected by clone formation test, changes in construction and form of Climbing film cells could be observed under the light microscope after 24 hours, and the percentage changes of apoptosis and the distributional percentage of the cell cycle were analyzed with flow cytometry. Results: Instant and 24-hour survival rates, growth rates, and capability of cell colony formation of A549 cells decreased more obviously in NA-combined NFU group than those in the other four groups. On the contribution of NA-combined NFU, the number of karyokinesis decreased significantly, while karyopyknosis and nuclear fragmentation increased greatly. Flow cytometry showed that the apoptotic index in NA-combined NFU group was higher than that in the NA or NFU group alone, and the cell cycle was held up in G1 stage. Conclusion: Certain concentration of NA has enhancement property on NFU and can increase the irradiated effectiveness of focused ultrasound markedly, which can provide reliable evidence for clinical application of NA as an ultrasound sensitizer.

Modulating activity of indomethacin to vincristine cytotoxicity in various human carcinoma cells.

The modulating activity of indomethacin to vincristine (VCR) was investigated in thirty human pulmonary carcinoma cell lines (adenocarcinoma 9, large cell carcinoma 9, squamous cell carcinoma 6, small cell carcinoma 6) and five other cell lines (colon carcinoma 2, melanoma 1, teratocarcinoma 1, thymoma 1). The survival of these cell lines treated with VCR with or without indomethacin (2 microg/ml) for 72 hours were examined using MTT assay, and IC50 values were calculated. When the cutoff level of potent combined effect in clinical use was set at 2-fold increase of sensitivity, the positive rate was 100% for adenocarcinomas and large cell carcinomas, 25% for squamous cell carcinomas, 33% for small cell carcinomas. Mean modulating index was 2.91 in adenocarcinomas, 1.92 in squamous cell carcinomas, 3.06 in large cell carcinomas and 1.67 in small cell carcinomas. Of the cell lines of other tumors, three cell lines (colon carcinoma 1, melanoma 1, teratocarcinoma 1) showed the potent combined effect of VCR and indomethacin, while indomethacin was not effective in modulating activity to VCR in a thymoma cell line and fibroblast cells. In conclusion, it is considered that modulating activity of indomethacin for VCR is a general effect for various human cancer cells, and combined use with VCR and indomethacin may be a useful modulation therapy for the advanced lung cancer.

Transcriptional and posttranscriptional mechanisms regulate human renin gene expression in Calu-6 cells.

We have recently identified a human pulmonary carcinoma cell line (Calu-6) that expresses human renin (hREN) mRNA endogenously, and we use it herein as a model to examine the regulation of the hREN gene. Transfection analysis of a deletion series (-2750 to -149) of hREN promoter-luciferase fusion constructs revealed the presence of multiple weak regulatory elements within the first 1,301 bp of the 5'-flanking region and a classic silencer element within the first intron (intron A) of the gene. The 5'-flanking regulatory domain consisted of three closely linked elements, two negative and one positive, each contributing a cell-specific threefold modulation of transcriptional activity. Treating Calu-6 cells with forskolin caused a 100-fold increase in steady-state endogenous hREN mRNA but no increase in hREN promoter activity in transient transfections or in nuclear runoff transcription assays. Nevertheless, de novo transcription and translation were necessary for adenosine 3',5'-cyclic monophosphate (cAMP)-mediated induction. Our results suggest that multiple regulatory elements regulate basal transcriptional activity of the hREN gene and the increase in hREN mRNA by cAMP may be mediated by posttranscriptional mechanisms.

Metabolic activation of 4-ipomeanol in human lung, primary pulmonary carcinomas, and established human pulmonary carcinoma cell lines : McLemore TL, Litterst CL, Coudert BP et al. 170 Eighth St. S.E., Paris, TX 75460. J Natl Cancer Inst 1990;82:1420–1426

Metabolic activation of 4-ipomeanol in human lung, primary pulmonary carcinomas, and established human pulmonary carcinoma cell lines : McLemore TL, Litterst CL, Coudert BP et al. 170 Eighth St. S.E., Paris, TX 75460. J Natl Cancer Inst 1990;82:1420–1426

Metabolic Activation of 4-Ipomeanol in Human Lung, Primary Pulmonary Carcinomas, and Established Human Pulmonary Carcinoma Cell Lines

4-Ipomeanol (IPO) is a pulmonary-specific toxin that is metabolically activated by a cytochrome P450 pathway in lung tissue. In this study, IPO metabolism, as determined by measurement of [14C]IPO covalent binding, was evaluated in a diverse sampling of 18 established, human lung cancer cell lines as well as in normal lung tissue and primary lung carcinoma tissue obtained at the time of thoracotomy from 56 patients with lung cancer. [14C]IPO covalent binding in lung cancer cell lines ranged from 248 to 1,047 pmol of bound [14C]IPO per milligram of protein per 30 minutes (mean +/- SE = 547 +/- 62.2). IPO metabolism in normal lung tissue ranged from 12 to 2,007 pmol of covalently bound [14C]IPO per milligram of protein per 30 minutes (mean +/- SE = 549 +/- 60). In lung cancer tissue, values ranged from 0 to 2.566 pmol of covalently bound [14C]IPO per milligram of protein per 30 minutes (mean +/- SE = 547 +/- 60, P greater than .3). When patients were divided into smokers and current non-smokers (no tobacco products smoked for greater than 6 mo), no effects of cigarette smoking were observed for either normal lung tissue or lung tumor tissue (P greater than .1 in all instances). A wide range of IPO metabolic activity was observed among different histological classifications of lung cancer cell lines and of fresh lung cancer tissues. IPO metabolism was simultaneously compared in normal lung tissue and lung cancer tissue from individual patients, but no positive correlation was observed (r = .10; P greater than .30). The results clearly demonstrate a wide range of IPO metabolism in both normal and lung cancer cells and indicate that a wide diversity of human lung cancers possess the metabolic enzyme system(s) necessary for the bioactivation of IPO to a potentially cytotoxic intermediate. Therefore, the continued exploration for any possible therapeutic potential of IPO in patients with lung cancer appears warranted.

Altered Regulation of the Cytochrome P4501A1 Gene: Novel Inducer-Independent Gene Expression in Pulmonary Carcinoma Cell Lines

The cytochrome P450 (CYP) systems catalyze the metabolic transformation of a wide variety of xenobiotics including procarcinogens present in cigarette smoke condensate as well as atmospheric pollutants. The CYP1A1 isoenzyme is of particular interest because it has been implicated as a risk factor in the etiology of lung cancer in heavy cigarette smokers. The identification and expression of the structural CYP1A1 gene in either normal human lung or lung cancer cells has not been reported. Because of its potential significance in human lung cancer, we investigated the expression of the CYP1A1 structural gene in 24 established human lung cancer cell lines including 15 non-small cell (eight adenocarcinomas, three large cell undifferentiated carcinomas, two bronchioloalveolar cell carcinomas, and two squamous cell carcinomas) and nine small cell lung carcinomas. CYP1A1 mRNA was detected in 14 of 15 (93%) of the non-small cell lung carcinoma cell lines examined following 24-hour treatment with benz[a]anthracene (BA) and in nine of 15 (60%) of the non-small cell lines cultured without an inducer in the medium. When the small cell lung cancer lines were evaluated for CYP1A1 gene expression, two of nine (22%) expressed detectable CYP1A1 mRNA in both BA-induced cell cultures and constitutive (control) cultures. A positive correlation was noted between BA-induced CYP1A1 mRNA levels and the corresponding aryl hydrocarbon hydroxylase activity expressed as absolute BA-induced enzyme activity (r = 0.74; P less than .01; n = 24), which further demonstrated that CYP1A1 mRNA expression reflects CYP1A1 enzyme activity in the individual cell lines. These observations represent the first known demonstration of constitutive (non-induced) CYP1A1 gene expression in human cells and suggest altered regulation of the CYP1A1 gene in selected lung cancer cell lines. These human pulmonary carcinoma cell lines, which have documented regulatory defects, could be useful for further identification of the mechanisms associated with CYP1A1 gene regulation.