Anticancer and antioxidant studies of the methanol extract and fractions of <i>Conyza sumatrensis</i> (retz.) E. H. Walker (asteraceae

In recent years, the increase in cancer morbidity and mortality has presented scientists with a major challenge in developing new therapeutic agents against cancer cells. This study aims to characterize the anticancer effects of copper oxide nanoparticles (NPs) conjugated with Lapatinib (CuO@Lapatinib) on breast and lung cancer cell lines. The physicochemical properties of the NPs were characterized by fourier-transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), scanning and transmission electron microscopy, energy dispersive X-ray spectroscopy (EDS), dynamic light scattering (DLS), and zeta potential analyses. The antiproliferative potential of the NPs in the breast (MDA-MB-231) and lung (A549) cancer cell lines and a normal cell line (MRC5) was investigated by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay. Flow cytometry and Hoechst staining were used to evaluate cell apoptosis and cell cycle analysis. The reactive oxygen species (ROS) levels in the treated and control cells were also determined. The NPs were spherical, with a size range of 20-59nm, a DLS size of 338nm, and a zeta potential of -42.9 mV. The half maximal inhibitory concentration (IC50) of CuO@Lapatinib NPs for the normal, breast cancer, and lung cancer cell lines was 105, 98, and 87 µg/ml, respectively. Treatment with CuO@Lapatinib NPs caused considerable apoptosis induction in breast cancer (from 0.65% to 68.96%) and lung cancer cell lines (from 1.11% to 44.11%). Also, an increased level of cell cycle arrest at the S phase was observed in both cancer cell lines. The ROS level in the breast and lung cancer cell lines after treatment with CuO@Lapatinib NPs increased by 3.45 and 21.04 folds, respectively. Nuclear morphological alterations, including chromatin condensation and fragmentation, were observed in both cancer cell lines. This study indicates CuO@Lapatinib is a potent antiproliferative compound with more efficient inhibitory effects on lung cancer than breast cancer cells, which can be related to the higher ROS generation in the A549 cell line.

1,3,5-triazine derivatives as potential anticancer agents against lung and breast cancer cell lines: Synthesis, biological evaluation, and structure-based drug design studies

Cancer cell lines have been, and will continue to be, important tools in oncology research, but there are major limitations in the fidelity of existing cell lines as a model of human cancers. Therefore, the aim of our study was to generate and characterize patient-specific cell lines from solid tumors that more faithfully recapitulate the primary tumor genetic and morphologic characteristics. We performed cell line derivation on a primary human breast adenocarcinoma (ER+/PR+/HER2-), a lung adenocarcinoma, a high-grade serous ovarian carcinoma, and an endometrioid ovarian carcinoma procured from a commercial tissue bank. Portions were snap-frozen for molecular analysis, and the remainder was minced and processed for cell culture. Cells were cultured and the resulting monolayer was stained by immunocytochemistry with a pan-keratin antibody for the presence of epithelial cells. Any contaminating fibroblasts were removed. We extracted DNA from the resulting breast and lung cancer cell lines at low, medium, and high passage numbers (approximately 20, 40, and 150 population doublings), as well as the corresponding primary tumor specimen, and a 250K SNP array was performed. These data were combined with that of nine breast cancer and 13 lung cancer cell lines from the GEO database, and subjected to genotyping analysis using the BLRMM algorithm in the Affymetrix Genotyping Console. The percent identity between the primary tumor tissue and the cell line was calculated from the number of identical SNP calls divided by the total number of SNP calls. Probesets resulting in “no call” genotyping results due to poor quality reads for either sample were not included in the analysis. For comparison, unrelated cancer cell line data of the same tumor type was included in the analysis. The cell lines reported here have grown for a minimum of 25 population doublings without evidence of cell culture crisis. The lung cancer, serous ovarian, and endometrioid ovarian cancer cell lines were positive for pan-keratin staining by immunocytochemistry. Both ovarian cancer subtypes exhibited tumorsphere formation under non-adherent culture conditions, whereas the lung cancer cell line formed loose aggregates. The percent identity between the primary carcinoma and cell line at 20, 40, and 150 population doublings was determined using a SNP call quality threshold of 0.1, resulting in an average 100K and 56K highest quality reads for breast and lung. The percent identity for the breast cancer cell line resulted in 98.7%, 97.4%, and 98.3% (mean +/- SD = 98.2% +/- 0.7%). The percent identity for the lung adenocarcinoma cell line at 20, 40, and 150 population doublings was 89.5%, 84.4%, and 87.2% (mean +/- SD = 87.0% +/- 2.5%). In comparison, the average % identity between unrelated existing breast cancer cell lines was 61.0% (+/- 3.1%) and between unrelated lung cancer cell lines was 58.7% (+/- 3.7%). These patient-specific breast and lung cancer cell lines exhibit genetically stable, extended growth from a primary tissue specimen for a minimum of 150 population doublings. We continue to measure extended growth in the ovarian cancer cell lines as well as a colon adenocarcinoma. Novel cell lines may be derived using this technology to represent specific patient groups and to serve the needs of researchers in precision medicine performing drug screening, target discovery, and the development of companion diagnostics. Citation Format: Naghmeh Salimi, Jeffrey D. Kent, Alex Chao, Bryan E. Roberts, Agoston Agoston, Elin S. Agoston. Patient-specific cancer cell line derivation methodology results in genetic stability across 150 population doublings. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr 33.

The primary objective of this research was to use flow cytometry to gain mechanistic insights into the cytotoxic effects of Tribulus terrestris extracts on breast cancer (MCF7) and lung cancer (A549) cell lines. T. terrestris was extracted using a Soxhlet apparatus in a progressive process. GC–MS was used to establish the phytochemical constituents. The amounts of phenolic compounds and flavonoids in the plant extracts were calculated using spectrophotometric analysis. The cytotoxicity of plant extracts was initially evaluated in non-malignant L929 cells, then in carcinogenic MCF-7 and A549 cell lines. Then, we performed an Annexin V assay, an anti-Bcl-2 assay, a Caspase-3 assay, and a DNA fragmentation (TUNEL) assay, using flow cytometry to investigate the underlying molecular processes. Based on the data, the methanolic extract of T. terrestris contained the highest amounts of phenolic compounds and flavonoids, with values of 169.87 µg GAE/g dwt and 160.12 µg QE/g dwt, respectively. Analysis by GC–MS revealed the presence of bioactive phytochemicals with proven cytotoxicity. Based on the MTT experiment, we determined that the IC50 values for the methanol extract’s effect on the viability of the MCF-7 and A549 cell lines were 218.19 and 179.62 µg/mL, respectively. The aqueous and methanol extracts were less cytotoxic when tested against the cancer-free L929 cell line (IC50 = 224.35 µg/mL). In both breast and lung cancer cells, the methanolic extract was found to activate caspase-3 and inhibit the Bcl-2 protein, resulting in early and late apoptosis and cell death via DNA damage. These findings point to cytotoxic effects of T. terrestris methanol extract against breast and lung cancer cell lines. Due to its potential as a source of anti-cancer chemotherapeutic medicines, T. terrestris warrants further investigation.

Current research in drug discovery from medicinal plants involves the identification of safe and inexpensive lead molecule towards disease tagets. Cancer is a second major national burden leading to cause death. Reducing the nation's cancer burden is a great challenge to win the “War of cancer”. During the last decades, numerous novel compounds have been found from marine organisms with interesting pharmaceutical activities. Hence, a novel brown algae, Sargassum ilicifolium (Turner) C.Aardh belonging to Sargassaceae family has selected and the study was aimed to investigate the in vitro anti-proliferative activity of Ethanolic extract and Chloroform fraction of Sargassum ilicifolium (Turner) C.Agardh. against Colon Cancer (HT-29) and Lung Cancer (A549) cell lines. Extraction of seaweed using 70% ethanol and fractionated with various solvents. Based on phytochemical screening, chloroform fraction was selected, then it was subjected to GC-MS analysis. The 70% ethanolic exract and chloroform fraction were selected to in vitro MTT cell line assay using Colon cancer cell line (HT-29) and lung Cancer cell line (A549). The GC-MS analysis report confirmed the presence of 21 compounds and in the the invitro assay, IC 50 value showed the effective anti-proliferative activity role against colon and lung cancer cell lines. The results of the present study proved the anti proliferative effect of Sargassum ilicifolium in the area of cancer. Further, the active biomolecule in the fraction has to be isolated , charecterized the formulated product subjected to in vivo study to strengthen the anticancer activity. Keywords: Antiproliferative effect, Colon cancer, lung cancer, Sargassum ilicifolium , GC-MS analysis.

Preclinical Studies of Gemcitabine and Trastuzumab in Breast and Lung Cancer Cell Lines

A synthesis of new derivatives of N'-(4H-benzo[b][1,4]thiazine-3-carbonyl) benzene sulfonyl-hydrazides. Characterized the final compounds using spectroscopic methods to ensure their synthesis and evaluation of their activity against two cancer cell lines; A504 human lung cancer cell lines and MCF-7 a human breast cancer cell line with estrogen, progesterone, and glucocorticoid receptors (i.e hormone-dependent). The synthesized compounds (S1, S2, S3, and S4) were successfully synthesized with a good yeild%, characterized using FTIR spectra, 1H NMR, and 13C NMR, and the assessment of their anti-proliferative activity against the two cancer cell lines by MTT method. There was an indication that the final compounds were prepared with a good yield using a precise research method. All derivatives promise anticancer activity for both cell lines; their half-maximal inhibitory concentration (IC50 in µM) for compounds (S1-S4) was 10.2, 5.4, 19.9, and 4.1 respectively against the A504 cell line, and 4.5, 5.4, 6, 9.3, and 2.6 respectively against MCF-7 cell line. The inhibition % of the final compounds (S1-S4) was 82.3, 88.3, 70.5, and 82.3 against the A504 cell line; and 75.9, 74.5, 72.7, and 80.4 respectively against the MCF-7 cell line. All final compounds were synthesized in a precise method in good yields, characterized spectroscopically showing successful synthesis, assessment of their anti-proliferative activity showed all of them have good IC50 and Inhibition % against A504 and MCF-7 cell lines; the compound S4 had the most promising one (IC50=4.18 and 2.68µM; inhibition percentage=82.3% and 80.45%, against A504 and MCF-7 respectively), compared with erlotinib and tamoxifen as references authorized anticancer drugs.

Phytochemicals from Corchorus olitorius methanolic extract induce apoptotic cell death via activation of caspase-3, anti-Bcl-2 activity, and DNA degradation in breast and lung cancer cell lines

BackgroundDespite recent advances in the treatment of lung and breast cancer, the mortality with these two types of cancer is high. Xanthohumol (XN) is known as a bioactive compound that shows an anticancer effect on cancer cells. Here, we intended to investigate the anticancer effects of XN on the breast and lung cancer cell lines, using the three-dimensional (3D) cell culture.MethodsXN was isolated from Humulus lupulus using Preparative-Thin Layer Chromatography (P-TLC) method and its authenticity was documented through Fourier Transform Infrared spectroscopy (FT-IR) and Hydrogen Nuclear Magnetic Resonance (H-NMR) methods. The spheroids of the breast (MCF-7) and lung (A549) cancer cell lines were prepared by the Hanging Drop (HD) method. Subsequently, the IC50s of XN were determined using the MTT assay in 2D and 3D cultures. Apoptosis was evaluated by Annexin V/PI flow cytometry and NFκB1/2, BAX, BCL2, and SURVIVIN expressions. Cell cycle progression was determined by P21, and P53 expressions as well as PI flow cytometry assays. Multidrug resistance was investigated through examining the expression of MDR1 and ABCG2. The invasion was examined by MMP2, MMP9, and FAK expression and F-actin labeling with Phalloidin-iFluor.ResultsWhile the IC50s for the XN treatment were 1.9 µM and 4.74 µM in 2D cultures, these values were 12.37 µM and 31.17 µM in 3D cultures of MCF-7 and A549 cells, respectively. XN induced apoptosis in MCF-7 and A549 cell lines. Furthermore, XN treatment reduced cell cycle progression, multidrug resistance, and invasion at the molecular and/or cellular levels.ConclusionsAccording to our results of XN treatment in 3D conditions, this bioactive compound can be introduced as an adjuvant anti-cancer agent for breast and lung cancer.

The insulin-like growth factors (IGFs), IGF-I and IGF-II, are potent mitogens for human lung and other epithelial cancer cell lines. Previous studies in defined medium lacking added IGF or insulin suggest that an IGF-related ligand can act as an autocrine growth factor for many cancer cell lines through action via the type I IGF receptor (IGF-R). Analysis of RNA isolated from human lung and breast cancer cell lines by reverse transcription of mRNA and polymerase chain reaction reveal that IGF-I and IGF-II mRNAs were co-expressed with IGF-R in the majority of cell lines. IGF-I mRNA was detected in 11/12 small cell lung cancer cell lines (SCLC), 13/14 nonsmall cell lung cancer (NSCLC) cell lines, and 1/2 breast cancer cell lines. IGF-II mRNA was detected in 8/10 SCLC, 11/12 NSCLC cell lines, and 2/2 breast lines. All cell lines expressed IGF-R. For analysis of IGF peptide secretion, cell lines were adapted to growth in serum/hormone-free culture medium (R0), and to avoid interference by IGF-binding proteins, secreted IGF peptides were isolated under acidic conditions and analyzed by Western blotting. Based upon measurement of the sensitivity of the anti-IGF antibodies for detection of recombinant human IGFs, IGF peptides accumulated in conditioned medium at greater than picomolar concentrations should have been readily detected. In three cell lines (two lung and one breast) secreted IGF immunoreactivity was detected as three molecular mass species of 23, 14, and 6 kDa. Isolation and NH2-terminal sequencing of each of these species definitively identified them as differentially processed forms of the IGF-II prohormone. Despite the high frequency of IGF-I gene expression detected by reverse transcription-polymerase chain reaction analysis, only one lung cancer cell line, NCI-N417d, was found that unequivocally secreted IGF-I peptide. This direct sequence determination unambiguously identifies IGF-II as the predominant IGF involved in the autocrine growth stimulation of human lung and breast epithelial tumor cell lines and supports a growing body of literature that implicates IGF-II/IGF-R autocrine loops as a common growth mechanism in epithelial carcinogenesis.

Chromosomal abnormalities and genomic instability in 10q22.3 region have been associated with lung cancer development, histopathological staging, and poor prognosis. Genetic mapping identified several genes including ZMIZ1, SFTPA1 and SFTPA2 in this critical region. Loss, interruption or gain of function of these genes in association with 10q22.3 deletions, translocations, or amplifications that were frequently detected in human primary lung cancer and cancer cell lines may promote oncogenesis. Human ZMIZ1, also named hZimp10, is a novel PIAS (protein inhibitor of activated STAT)-like protein that shares a zing finger domain, MIZ (MSX-interacting zinc finger), with other PIAS proteins. Fusion of ZMIZ1 to ABL1 with a t(9;10)(q34;q22.3) translocation was identified in a B-cell acute lymphoblastic leukemia patient. In this study, we performed fluorescence in situ hybridization (FISH) using genomic DNA probes covering ZMIZ1 and SFTPA gene loci at 10q22.3 to determine chromosomal aberrations involving the ZMIZ1 gene in primary lung tumor samples and lung cancer cell lines. Chromosomal abnormalities at 10q22.3 loci, including deletions, polysomy, and translocations were frequently detected in tumors and tumor-derived cell lines. We analyzed copy number variations specifically linked to ZMIZ1 gene loci by mining the published genome-wide SNP dataset from 370 NSCLC tumor samples and confirmed a significant copy number gain with average copy numbers (CN) > 5 or loss with CN < 1.5, respectively. A high frequency of loss of heterozygosity (LOH) of ZMIZ1 and ABL1 genes in lung cancer cell lines and high level amplifications of ZMIZI gene (CN > 7) in breast and prostate cancer cell lines were also detected in Sanger SNP and CGP databases, respectively. We performed Spectral karyotyping (SKY) analysis to identify chromosomal abnormalities and potential translocations involved in the 10q region in lung cancer cell lines. We found that the 10q region was translocated and fused with other chromosomes extensively in H1819 cells. This result was consistent with the high frequency 10q chromosomal abnormalities detected in human lung and other cancer cell lines in the NCI/NCBI SKY/M-FISH & CGH database. Furthermore, we developed and performed a high-throughput RACE-PCR-based gene-specific RNA sequencing analysis using next-generation parallel sequencing technology (454 Life Sciences, Inc.) to search for potential transcriptional variants and fusion partners of the ZMIZ1 gene in lung cancer cell lines which has revealed potential fusion mRNA. These results suggest that ZMIZ1 may play an important role in lung cancer development and serve as a useful biomarker for lung cancer molecular staging and prognosis. (Supported by NIH/NCI grants SPORE P50CA70907 and RO1CA116322) Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1177.

The present study aimed to perform phytochemical investigation of various fractions of the aerial parts of Senecio laetus Edgew for its bio-constitution and to pharmacologically evaluate these fractions for antioxidant and antiproliferative activities against breast cancer (MCF-7) and colon carcinoma (HCT-116) cell lines. Phytochemical screening and characterization of compounds was carried out as per standard procedures. Extracts were subjected to GC-MS analysis for identification and characterization of constituents. The antioxidant activity was carried out using DPPH, ferric ion reducing power assay, and nitric oxide radical inhibition assays. In vitro antiproliferative activity of the extracts was carried out using MCF-7 (breast cancer) and HCT-116 (colon carcinoma) cancer cell lines by MTT assay, colony formation assay, and wound healing assay. Phytochemical screening showed the presence of alkaloids, flavonoids, carbohydrates, glycosides (cardiac and anthraquinone), tannins, triterpenoids, phenolic compounds, and phytosterols. GC-MS analysis showed the presence of 54 compounds. Ethyl acetate and methanolic extract exhibited the maximum amount of phenolic content compared to dichloromethane and hexane fractions. Antioxidant capacities were shown highest in ethyl acetate and methanolic fractions. The antiproliferative activity was found to be concentration dependent, with dichloromethane fraction more effective than ethyl acetate and hexane fractions against both the MCF-7 and HCT-116 cancer cell lines. The results indicate that S. laetus Edgew has a promising antioxidant and antiproliferative potential as shown by its activity against MCF-7 (breast carcinoma) and HCT-116 (colon carcinoma) cancer cell lines.

Objective: To investigate the in vitro cytotoxic activity of pigment extracted from Salinicoccus sp. isolated from Nellore sea coast. Materials and Methods: In the present study, pigment-forming bacteria were isolated from samples collected from Nellore coast, Andhra Pradesh. Among different pigmented isolates obtained on Zobell agar medium, the pinkish orange bacterium was selected for the study. The bacterium was cultured in Zobell broth medium and incubated in an orbital shaker at 120 rpm for 6 days at 25°C. After incubation, the culture broth was centrifuged at 10,000 rpm for 10 min to obtain a pellet, and the pellet was extracted using the solvent methanol and acetone (5:1). The crude pigment extract was evaluated for cytotoxic potential and was found to exhibit cytotoxic effect on A549 (human lung cancer) and MCF-7 (breast cancer) cell lines. The cell lines A549 cells, MCF-7, were cultured in Dulbecco's Modified of Eagle medium with L-glutamine and 1000 mg/L glucose supplemented with 10% fetal bovine serum, penicillin G (100 units/ml), and streptomycin sulphate (0.1 mg/ml) in a humidified atmosphere consisting of 5% CO2at 37°C. Results: The results of the present study revealed that the crude pigment extract has a strong anticancer potential, especially toward the A549 (Lung cancer cell lines) and MCF-7 (Breast cancer cell lines), respectively. Conclusion: This study clearly indicated that the pigment extract of marine Salinicoccus sp. has a strong cytotoxic activity against A549 (lung) and MCF-7 (breast) cancer cell lines which may be utilized for the drug development.

The objectives of this research were to carry out GC–MS and LC–MS-based phytochemical profiling of Barleria hochstetteri, as well as flow cytometry-based mechanistic investigations of the cytotoxic effect of its extracts against breast and lung cancer cell lines. This preclinical in vitro study was carried out in Saudi Arabia and India, from 11 August to 15 January 2022. Barleria hochstetteri was sequentially extracted using the Soxhlet extraction technique. Utilizing LC–MS and GC–MS methods, the phytochemical profiling was performed. Additionally, the total phenolic compounds and flavonoids were quantified in the plant extract using spectrophotometric techniques. In this study, we first examined the cytotoxicity of the plant extract on non-malignant L929 cells and on the carcinogenic MCF-7 and A549 cell lines. Then, we studied the underlying molecular pathways by means of Anti-Bcl-2, caspase-3, and DNA fragmentation (TUNEL) assays, using flow cytometry. The results revealed phenolic compounds and flavonoids to be the two major components in the methanolic extract of B. hochstetteri, with concentrations of 3210 µg GAE/g dwt and 1863 µg QE/g dwt, respectively. Results from GC–MS and LC–MS analyses revealed the presence of bioactive phytochemicals with known cytotoxicity. From the MTT assay on cell viability, the IC50 of the methanol extract for the MCF-7 and A549 cell lines were 219.67 and 144.30 µg/mL, respectively. With IC50 values of 324.24 and 266.66 µg/mL, respectively, the aqueous and methanol extracts were less toxic when tested against the non-cancerous L929 cell line. The extract caused early and late apoptosis in the tested breast and lung cancer cells by activating caspase-3 and inhibiting Bcl-2 protein, and it also caused cell death via DNA damage, based on flow cytometric and molecular marker analyses. These findings indicate that the methanol extract of B. hochstetteri was cytotoxic on breast cancer and lung cancer cell lines. To uncover cancer-fighting chemicals, there is a need for further research on B. hochstetteri, as it is a promising source of anti-cancer chemotherapeutic drugs.

Exploring the anticancer potential of Cytotoxin 10 from Naja kaouthia venom: Mechanistic insights from breast and lung cancer cell lines