Human Solid Cancer Cell Lines Research Articles

Abstract 2424: CD40 expression in human solid tumors and NCI60 tumor cell lines.

Abstract Cell-surface CD40 is a member of the tumor necrosis factor receptor superfamily (TNFRSF) expressed and functionally well characterized in immune and hematopoietic cells. However, its expression and role in human solid tumors remain to be evaluated. As a molecular target currently with the antibody-based cancer therapy, it is important to characterize expression of CD40 across a range of human solid tumors and cancer cell lines. This could help improve our understanding of biology of CD40 molecule in human cancers especially epithelial tumors, and in turn increase the odds of success of targeting CD40 pathway. Expression of CD40 in the NCI60 human tumor cell lines and common types of human solid tumors were examined by immunohistochemistry in formalin-fixed and paraffin-embedded tumor specimens. CD40, predominantly membranous signal, was constitutively expressed in 15 lines, respectively derived from carcinomas of the breast, colorectum, non-small cell lung, ovary, prostate, and renal cell, of the NCI60 tumor cell line panel, and confirmed by Western blotting and gene expression microarray data available at http://dtp.nci.nih.gov/mtweb/search.jsp. CD40 expression was largely in accordance with the mRNA expression data measured by gene expression profiling. The status of CD40 was verified in MDA-MB-231, T-47D and HCT-116 cells (CD40-positive), and MCF-7 and HT-29 cells (CD40-negative) by Western blot. Moreover, CD40 was detected in tumor cells in 17% (6/34) of breast cancer, 24% (8/34) colorectal cancer, 54% (23/43) non-small cell lung cancer, and 47% (17/36) ovarian cancer. CD40 was also spotted in infiltrating lymphocytes in the tumor stroma. The data indicate that CD40 is expressed in human solid tumors with varying expression frequencies and one quarter of cancer cell lines examined. In addition, we have established a ready-for-use immunohistochemistry assay with an antibody that specifically detects CD40 in formalin-fixed and paraffin-embedded human tumor samples. Its application holds promise to evaluate the prognostic role of CD40 in human cancers. Citation Format: Dat Nguyen, Joseph E. Tomaszewski, Stephen M. Hewitt, James H. Doroshow, Sherry X. Yang. CD40 expression in human solid tumors and NCI60 tumor cell lines. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2424. doi:10.1158/1538-7445.AM2013-2424

Anti-proliferative effect of (19Z)-halichondramide, a novel marine macrolide isolated from the sponge Chondrosia corticata, is associated with G2/M cell cycle arrest and suppression of mTOR signaling in human lung cancer cells

Five oxazole-containing macrolides isolated from the marine sponge Chondrosia corticata were evaluated for their anti-proliferative activity in a panel of human solid cancer cell lines. (19Z)-Halichondramide ((19Z)-HCA), a novel trisoxazole-containing macrolide, exhibited the highest potency among the macrolides, with IC50 values in the submicro-molar ranges. Prompted by the high potency of growth inhibition of cancer cells, we investigated the mechanism of action of the anti-proliferative activity of (19Z)-HCA in human A549 lung cancer cells. (19Z)-HCA induced cell cycle arrest in the G2/M phase, and this event was highly correlated with the expression of checkpoint proteins, including the up-regulation of p53 and GADD45α and the down-regulation of cyclin B1, cyclin A, CDC2, and CDC25C. In addition, the growth inhibition by (19Z)-HCA was associated with the suppression of mTOR and its downstream effector molecules 4EBP1 and p70S6K. The modulation of mTOR signaling by (19Z)-HCA was found to be mediated by the regulation of upstream proteins, including the down-regulation of Akt and p38 MAPK and the up-regulation of AMPK. These data suggest the potential of (19Z)-HCA to serve as a candidate for cancer chemotherapeutic agents derived from marine organisms by virtue of arresting the cell cycle in the G2/M phase and the modulation of mTOR/AMPK signaling pathways.

Cytotoxic effect of Mercurialis annua L. methanolic extract on six human solid cancer cell lines

The unprecedented rates of resistance of cancer cells to traditional chemotherapeutic medications necessitated relentless exploration and drug design in an attempt to introduce safe and effective new antineoplastic agents. The latter is a particular pivotal to the treatment of solid tumors malignancies which represent the top causes of cancer deaths. Our laboratory has established a line of research to investigate potential cytotoxic agents of native plant origins. Folkloric anecdotal evidence and initial screening led us to investigate the cytotoxicity of Mercurialis annua L. (Euphorbiaceae) methanolic extract against a variety of cancer cell lines. In total, six different cell lines representing three types of common cancers were used to determine the potential antineoplastic activity of the plant extract. Our results show that the M. annua extract exhibited potent cytotoxic effect on both HRT-18 and T47D cell lines in a dose-dependent manner. HRT-18 is a colorectal adenocarcinoma cell line, while T47D is a breast cancer cell line. The calculated IC50 values of the plant extract against HRT-18 and T47D cell lines were 41 and 270 µg/ml, respectively. On the other hand, there was no appreciable inhibitory effect of the plant extract on the growth of Caco-2, MCF-7, A375-S, and WM136-1A cell lines. The inhibition of cellular growth was cell-type specific. Among the breast cancer cell lines, MCF7 and T47D, only T47D which is a human ductal breast epithelial was sensitive to the plant extract. Likewise, the human epithelial colorectal adenocarcinoma cells (Caco-2) was resistant, while the human rectum adenocarcinoma (HRT-18) was sensitive to the plant extract. Interestingly, the two human malignant melanoma cell lines; A375.S2 and WM136 1A were both resistant to the plant extract. This work is a first step towards unraveling the full pharmacologic potential of M. annua in cancer treatment. Key words: Mercurialis annua L., cancer, cytotoxic, T47D, HRT-18, plant extract.

Anti-cancer mechanism of bufalin

Bufalin is one of the major active components of Chan Su,a traditional Chinese medicine.Studies at home and abroad have confirmed that bufalin can inhibit cell proliferation significantly in many human leukemia and solid cancer cell lines by inhibiting endothelial hyperplasia and angiogenesis,and inducing cell differentiation and apoptosis. With the deepening of study,it is showed that bufalin has extensive anticancer activities and it has potential clinical value with very low drug concentration. Key words: Bufalin ; Neoplasms ; Apoptosis

Identification of two small molecule inhibitors of hypoxia-inducible factor 1 with different cell-based screening model

To investigate the inhibitory activity of HIF-1 by triptolide and manasaantin A, two cell-based models with luciferase report gene assay were established. Two cell-based models of HIF-1 were used to evaluate HIF-1 inhibition activity of triptolide and manasaantin A. Secreted VEGF expression induced by hypoxia was detected by ELISA with two compounds. The growth inhibition of different solid tumor cell lines was measured by the MTT assay. The expression of firefly luciferase was induced by hypoxia in U251-HRE and T47D-HRE cells. U251-HRE model was suitable for the detection of HIF-1 inhibition activity of triptolide. The IC50 of triptolide on HIF-1 activity was (3.4 +/- 0.5) x 10(-8) mol x L(-1). The report gene assay using T47D cells co-transfected with pGL2-TK-HRE and pRL-CMV showed more sensitive inhibition activity of HIF-1 on manassantin A than that of detected by U251-HRE model. The IC50 of manassantin A on HIF-1 activity was (2.4 +/- 0.6) x 10(-8) mol x L(-1). HIF-1 target gene VEGF was also inhibited by test compounds on protein level in T47D cells. Manasaantin A showed selective inhibition on the growth of human solid cancer cell lines, especially on breast cancer and pancreatic cancer cells. Meanwhile, triplotide showed strong proliferation inhibition activity on all tested cell lines. It is very important to select a suitable cell-based report gene assay of HIF-1 for screening of different kinds of inhibitor.

Papuamine and haliclonadiamine, obtained from an Indonesian sponge Haliclona sp., inhibited cell proliferation of human cancer cell lines

The extract of an Indonesian marine sponge Haliclona sp. showed potent cytotoxicity against human solid cancer cell lines, MCF-7 (breast), LNCap (prostate), Caco-2 (colon) and HCT-15 (colon) cells. Study on nuclear morphological changes and flow cytometric analysis suggested that the component(s) in the extract would induce an apoptosis to these cancer cells. Bioassay-guided isolation yielded two pentacyclic alkaloids, papuamine (1) and haliclonadiamine (2), which inhibited cell proliferation of six human cancer cell lines with IC50 values of 0.93–1.50 and 1.00–4.44 µM, respectively. Compounds 1 and 2 accumulated lymphoma U937 cells at sub-G1 phase and induced a condensation of chromatin and fragmentation of nucleus.

Abstract 5328: Interleukin-4 receptor regulates metastasis of mammary epithelial cancer cells

Abstract Interleukin-4 (IL-4), a cytokine produced mainly by immune cells, may promote cancer cell survival by mediating apoptosis resistance in epithelial tumors. The IL-4 Receptor (IL-4R) is overexpressed in a variety of human solid cancer cell lines and primary cell lines, and may be a promising target for anti-tumor therapy. Here, we show that loss of IL-4Rα, a component of the IL-4 receptor, in two murine mammary tumor cell lines (R221A and 4T1), reduces tumor cell survival and results in attenuated metastatic growth in secondary sites. Proliferation and direct cell counting analyses, following knockdown of IL-4Rα with RNA interference, show that R221A clones with reduced IL-4Rα expression proliferate at a comparable rate to control clones, but the number of cells is significantly reduced. This data correlates with the finding that reduced IL-4Rα expression in R221A cells almost completely abolishes tumor take after orthotopic mammary injection in vivo. Furthermore, reduced IL-4Rα expression in R221A and 4T1 cell lines results in a decrease [2.4 fold] in lung metastatic growth in vivo. Reduced expression of IL-4Rα in 4T1 cells results in a similar decrease of liver metastasis. In support of this data, proliferation assays following serum starvation and a modified clonogenic survival assay show that both 4T1 and R221A knockdown clones have a decrease [2 fold] in survival ability in comparison to control clones. Taken together, our results indicate that the IL-4 receptor promotes survival of breast cancer cells at metastatic sites, and suggests that drugs targeting this pathway, currently in clinical trials for respiratory ailments, may have potential for limiting metastatic disease. 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 5328. doi:1538-7445.AM2012-5328

Kinase inhibitory potencies and in vitro antiproliferative activities of N-10 substituted pyrrolo[2,3-a]carbazole derivatives

Development of potent and selective Pim kinase inhibitors has recently emerged as an important field for the design of new anti-cancer drugs. We report the synthesis of new N-10-substituted pyrrolo[2,3-a]carbazole derivatives and their evaluation as Pim kinase inhibitors. Moreover, in vitro antiproliferative activity of these compounds was evaluated toward a human fibroblast primary culture and three human solid cancer cell lines (PA1, PC3 and DU145). Compounds 3, 7 and 10 showed inhibitory potencies toward Pim-1 and Pim-3 in the nanomolar range. Additionally, dimethylamino analog 10 also demonstrated interesting sub-micromolar antiproliferative activities toward the cell lines tested.

Antiproliferative activity of withanolide derivatives from Jaborosacabrerae and Jaborosa reflexa. Chemotaxonomic considerations

Three withanolides were isolated from the aerial parts of Jaborosa reflexa Phil. Jaborosa cabrerae Barboza yielded five sativolide withanolides (including jaborosalactones R, S, 38, and 39) and two trechonolide withanolides epimeric at C-23 (trechonolide A and jaborosalactone 32). In addition, five derivatives were obtained by chemical derivatization of jaborosalactone 38, and all compounds were fully characterized by 1D and 2D NMR spectroscopic studies. The in vitro antiproliferative activities of the major natural withanolides and the semisynthetic derivatives were examined against HBL-100, HeLa, SW1573, T-47D, and WiDr human solid tumor cancer cell lines. Some chemotaxonomic considerations are discussed.

QSAR studies on Withanolide analogs for anticancer activity

AbstractWithanolides are a group of pharmacologically active compounds present in most prodigal amounts in roots and leaves of Withania somnifera (Indian ginseng), one of the most important medicinal plants of Indian systems of medicine. Withanolides are basically steroidal lactones (highly oxygenated C-28 phytochemicals) and similar to ginsenosides activity. Some of the withanolides that have been reported possess immunomodulatory, and anticancer activities. In the present investigation, a quantitative structure activity relationship (QSAR) model based on forward stepwise multiple linear regression (MLR) has been developed against the MCF7, MCF7/BUS, and SK-Br-3 human solid tumor breast cancer cell lines. Relationship correlation coefficient (r2) and cross validation correlation coefficient (r2CV) of QSAR model were 0.77 and 0.73 for MCF7, 0.91 and 0.85 for MCF7/BUS, 0.93 and 0.90 for SK-Br-3 respectively. Developed QSAR model was also evaluated for prediction accuracy through internal, external and randomization validation methods. The QSAR study indicates that chemical descriptors viz., atom count (all atoms), connectivity index (order 2, standard), for MCF7, Connectivity Index (order 0, standard), Dipole Vector X (debye), Molar Refractivity, Shape Index (basic kappa, order 2) for SK-Br-3 and Atom Count (all atoms), Dielectric Energy (kcal/mole), Total Energy (Hartree), Heat of Formation (kcal/mole) for MCF7/BUS are correlate well with the breast cancer activity, Moreover, on the basis of screening for oral bioavailability, in silico ADME and toxicity risk assessment, we concluded that compounds W3, W4, W8 have markedly higher anticancer activity compared to control. These results can offer useful references for directing the molecular design of lead compound(s) based on withanolide or analogous template with improved activity.

Screen for natural benzylisoquinoline alkaloids against tumor

To search for natural benzylisoquinoline alkaloids against tumor. In this study, taken DNA as target, a method was developed for screening of natural benzylisoquinoline alkaloids against tumor from traditional Chinese medicine by the use of centrifugal ultrafiltration combined with HPLC technology. The anti-tumor activity for the compounds screened was evaluated in vitro. Nine compounds interacted with DNA were discovered and identified from Macleaya cordata, Chelidonium majus, Coptis chinensis, and the proliferation of four types of human solid cancer cell lines was markedly inhibited by these compounds. The developed method was considered to be suitable for screening of natural benzylisoquinoline alkaloids against tumor from traditional Chinese medicine.

Design, synthesis and in vitro drug release investigation of new potential 5-FU prodrugs

In order to identify new efficient prodrugs of 5-fluorouracil (5-FU) and to develop an original targeting approach using 2-fluoro-2-deoxyglucose (FDG) as a potential drug carrier, eight original 5-FU derivatives were synthesized: 5-FU was attached by the N1 position of the pyrimidinic ring to the C1 position of the FDG structure either by direct coupling ( 2a) or via various spacers ( 3, 6a– c, 10b and 19). A new sensitive high-performance liquid chromatography method was developed to simultaneously quantify 5-FU and its derivatives in human plasma and other relevant media at physiological temperatures. Half-lives were determined from the degradation profiles of these conjugates. Slow degradation of compounds 2a, 3, 10b and 19 was observed in vitro at 37 °C, but no 5-FU release was noticed. By contrast, the in vitro drug release profiles of compounds 6a– c followed pseudo-first-order kinetics, and 5-FU was found in all the media. The antiproliferative activity of the eight compounds was assessed in vitro by a fluorometric assay against two human solid cancer cell lines and one healthy cell line. A correlation was found between the activities of the compounds and their ability to release 5-FU efficiently.

Synthesis and biological activities of pyrazolo[3,4- g]quinoxaline derivatives

The synthesis of new pyrazolo[3,4- g]quinoxaline derivatives, as well as their Pim kinases (Pim-1, Pim-2, Pim-3) inhibitory potencies and in vitro antiproliferative activities toward a human fibroblast primary culture and three human solid cancer cell lines (PA1, PC3 and DU145) are described. The results obtained in this preliminary structure–activity relationship study have pointed out that most of the compounds in this series exhibited interesting in vitro Pim-3 kinase inhibitory potencies. Moreover, some of the tested compounds have demonstrated favorable antiproliferative potencies.

Synthesis, Pim kinase inhibitory potencies and in vitro antiproliferative activities of diversely substituted pyrrolo[2,3-a]carbazoles

The synthesis of new pyrrolo[2,3-a]carbazole derivatives diversely substituted at the C-6 to C-9 positions is described. These compounds were tested for their kinase inhibitory potencies toward three kinases (Pim-1, Pim-2, Pim-3) as well as for their in vitro antiproliferative activities toward a human fibroblast primary culture and three human solid cancer cell lines (PC3, DU145, and PA 1). Moreover, molecular docking studies were performed to explain the enhanced inhibitory activity of the most active compound 3d.

Antiproliferative Activity of Withanolides against Human Breast Cancer Cell Lines

The in vitro antiproliferative activity of a series of 22 naturally occurring withanolides was examined against the T-47D, MCF7, MCF7/BUS, MDA-MB-231, and SK-Br-3 human solid tumor breast cancer cell lines. The most active compound showed GI(50) values in the range 0.16-0.71 muM. The aromatic withanolide 19 exhibited specific activity for the estrogen-receptor-positive cell lines (T-47D, MCF7, and MCF7/BUS). Overall, the results demonstrated the relevance of the substitution pattern on the A and B rings on the resultant antiproliferative activity.

Synthesis and in vitro antiproliferative activities of quinoline derivatives

The synthesis of new di- and trimeric quinoline derivatives is described as well as their in vitro antiproliferative activities toward a human fibroblast primary culture and two human solid cancer cell lines (MCF-7 and PA 1).

Synthesis, Kinase Inhibitory Potencies, and in Vitro Antiproliferative Evaluation of New Pim Kinase Inhibitors

Members of the Pim kinase family have been identified as promising targets for the development of antitumor agents. After a screening of pyrrolo[2,3-a]- and [3,2-a]carbazole derivatives toward 66 protein kinases, we identified pyrrolo[2,3-a]carbazole as a new scaffold to design potent Pim kinase inhibitors. In particular, compound 9 was identified as a low nM selective Pim inhibitor. Additionally, several pyrrolo[2,3-a]carbazole derivatives showed selectivity for Pim-1 and Pim-3 over Pim-2. In vitro antiproliferative activities of 9 and 28, the most potent Pim inhibitors identified, were evaluated toward three human solid cancer cell lines (PA1, PC3, and DU145) and one human fibroblast primary culture, revealing IC50 values in the micromolar range. Finally, the crystal structure of Pim-1 complexed with lead compound 9 was determined. The structure revealed a non-ATP mimetic binding mode with no hydrogen bonds formed with the kinase hinge region and explained the selectivity of pyrrolo[2,3-a]carbazole derivatives for Pim kinases.

Synthesis and biological activities of aminopyrimidyl-indoles structurally related to meridianins

The synthesis of new meridianin derivatives substituted at the C-5 position of the 2-aminopyrimidine ring by various aryl groups and substituted or not by a methyl group on the indole nitrogen is described. These compounds were tested for their kinase inhibitory potencies toward five kinases (CDK5/p25, CK1δ/ε, GSK-3α/β, Dyrk1A and Erk2) as well as their in vitro antiproliferative activities toward a human fibroblast primary culture and two human solid cancer cell lines (MCF-7 and PA 1).

PI3K/Akt is involved in bufalin-induced apoptosis in gastric cancer cells

Bufalin is the active ingredient of the Chinese medicine Chan Su, and it has been reported that bufalin induces apoptosis in some human leukemia and solid cancer cell lines. The exact mechanism of bufalin-induced apoptosis is, however, still not clear. In this study, we demonstrated that bufalin inhibited the proliferation of gastric cancer MGC803 cells in a dose-dependent and time-dependent manner. At a low concentration (20 nmol/l), bufalin induced M-phase cell cycle arrest, whereas at a high concentration (80 nmol/l) it induced apoptosis in MGC803 cells. Bufalin increased the Bax/Bcl-2 ratio and activated caspase-3 during the apoptotic process of MGC803 cells. It should be noted that bufalin transiently activated the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway and then inhibited it completely, and upregulated the Casitas B-lineage lymphoma (Cbl) family of ubiquitin ligases, upstream modulators of PI3K. A combination of bufalin and LY294002, a PI3K-specific inhibitor, enhanced apoptosis, but PD98059, an extracellular-regulated protein kinase-specific inhibitor, had no significant effect on bufalin-induced apoptosis. These results suggested that the PI3K/Akt pathway might play a key role in bufalin-induced apoptosis in gastric cancer MGC803 cells.

In-vitro antiproliferative activities and kinase inhibitory potencies of meridianin derivatives

Marine alkaloid meridianin G derivatives, substituted on the pyrimidine ring by aryl groups, were evaluated for their kinase inhibitory potencies and their in-vitro antiproliferative activities. The derivatives were tested toward a panel of nine protein kinases (KDR, IGF-1R, c-Met, RET, c-Src, c-Abl, PKA, CDK2/cyclin A, and HER-1) and their in-vitro antiproliferative activities were evaluated toward a human fibroblast primary culture and two human solid cancer cell lines (MCF-7 and PA 1). Despite weak kinase inhibitory potencies, high in-vitro antiproliferative activities were found for compounds 5, 7, 12, and 14, which do not interfere with the PA 1 cell cycle and may be considered as direct cytolysis or apoptosis inducers.