Effect of SMAC Gene on Sensitivity of Lung Adenocarcinoma Cells to Paclitaxel and Cell Viability Based on caspase-3/Bcl-2/Bax Signaling Pathway / 肿瘤防治研究

Objective To investigate the effect of the SMAC gene on paclitaxel sensitivity and cellular activity in lung adenocarcinoma cells based on the caspase-3/Bcl-2/Bax signaling pathway. Methods A paclitaxel-resistant cell line A549/Taxol was established for lung adenocarcinoma, and the cells were divided into four following groups: pcDNA-NC (transfected with pcDNA-NC blank vector), pcDNA-SMAC (transfected with pcDNA-SMAC vector), siRNA-NC (transfected with siRNA-NC empty virus vector), and siRNA-SMAC groups (transfected with siRNA-SMAC lentiviral vector). The SMAC mRNA expression in cells was detected by qRT-PCR; cell sensitivity was detected by MTT; cell proliferation ability was detected by cloning assay; cell invasion ability was detected by Transwell; apoptosis ability was detected by flow cytometry assay; and caspase-3, Bcl-2 and Bax protein expression in cells were detected by Western blot analysis. Results The SMAC mRNA expression was significantly lower in A549 cells compared with BEAS-2B cells (P < 0.05). The SMAC mRNA expression was significantly higher in the pcDNA-SMAC group than that in the pcDNA-NC group cells (P < 0.05). The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P < 0.05) than that in the siRNA-NC group. The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P < 0.05) than in the siRNA-NC group. Compared with the pcDNA-NC group, the cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly lower in the pcDNA-SMAC group, the cell resistance index reversal was 2.51-fold, and the apoptosis ability and caspase-3, as well as Bax protein expression, were significantly higher (P < 0.05). Compared with the siRNA-NC group, cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly higher in the siRNA-SMAC group, and apoptosis ability and caspase-3 and Bax protein expression were significantly lower (P < 0.05). Conclusion High expression of SMAC increases paclitaxel sensitivity, inhibits cell growth and invasion, promotes apoptosis in lung adenocarcinoma cells, and has a regulatory effect on the caspase-3/Bcl-2/Bax signaling pathway.

Design, synthesis, and cytotoxicity evaluation of new 2,4-disubstituted quinazolines as potential anticancer agents

A series of new 2, 4-disubstituted quinazolines were synthesized by an analog design approach. The synthesis oftitle compounds (3a–f, 4a–c, 5a–c, and 6a–c) was achieved from the corresponding key intermediates 2-(pyridin3-yl) quinazolin-4(3H)-one(2a), 2-(pyridin-3-yl) quinazolin-4(3H)-one (2b) and 2-(pyrazin-2-yl)quinazolin-4(3H)-one (2c) with appropriate amines. The synthesized compounds were characterized by the spectral studies. All thesynthesized compounds were evaluated for in vitro anticancer activity employing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay against human adenocarcinoma (HT-29), breast cancer (MDA-231), and Ehrlichascites carcinoma cell lines. Among the tested compounds, 5a has a significant anticancer activity (5.33 µM/ml)against the human adenocarcinoma cell line. Other compounds have shown a moderate anticancer activity against thetested cell lines.

Homoeopathic Viscum album (10-3) is More Cytotoxic to in vitro Culture of Human Breast Cancer Cell Line than to Human Mesenchymal Stem Cells

BackgroundBreast cancer has been considered a public health problem and homeopathic treatments are becoming increasingly recommended due to its ways of action and absence of adverse effects. MCF-7 is an adenocarcinoma of human breast cell line useful as preclinicalmodel to screen therapeutic agents such as ultra-diluted Viscum album, an European plant which extract is commonly used in cancer therapy. AIMS MCF-7 and mesenchymal stem cells (MSC) were used to evaluate the in vitrocytotoxicity of homoeopathic Viscum album 1x10-3(VA3X). Methodscells were cultured for 24 hours in controlled environment (37.5oC and 5% CO2) in 96-well plates. After this time, VA3X was added to the culture medium in concentrations varying from 10 to 100 L/mL.A control group was maintained with culture medium only. Cells were cultivated for 48 hours in these conditions for evaluation of cell viability by MTT assay. ResultsHigher cytotoxicity was observed in MCF-7 when compared to MSC, as the lower concentration of VA3X was capable of inducing tumor cell death and not healthy cell death. The MTT assay results were that 42 L/mL of VA3X reduced MCF-7 cells viability to 50% and 62 L/mL reduced MSC cells to the same percentage, what means that tumor cells are more sensible to VA3X than heathy cells. ConclusionViscum albumpresented higher cytotoxic action on human breast cancer cell line culture than on mesenchymal stem cells. This medicine is extensively used against cancer, and the use of the homoeopathic form of it brings new possibilities as no or fewer adverse effects would be present.(AU)

Expression and function of advanced glycosylation end product receptors and intracellular signal molecules in lung adenocarcinoma cells / 安徽医科大学学报

Objective To explore the expression of the receptor for advanced glycosylation end products(RAGE) and its intracellular signaling molecules DIAPH1 in lung adenocarcinoma A549 cells and the effect of RAGE ligands on cell migration and apoptosis. Methods The expressions of RAGE and DIAPH1 in lung adenocarcinoma A549 cells and human bronchial epithelial cells BEAS-2B were tested by qRT-PCR and Western blot. A549 cells was treated with 10,100 μg /ml RAGE ligands CML-AGE and 1,10,100 μg /ml S100B,and wound healing test was used to identify the effect of migration ability. A549 cells was treated with 25,50,100 μg /ml RAGE ligands CMLAGE, the gene expression of BCL-2 and BAX were tested by using qRT-PCR. Results The results of qRT-PCR and Western blot showed,compared with human bronchial epithelium cells BEAS-2B,the expression of RAGE and DIAPH1 were both significantly down-regulated in lung adenocarcinoma A549 cells (P < 0. 001). After treated with 10,100 μg /ml RAGE ligands CML-AGE and 1,10,100 μg /ml S100B ,both groups showed the ligands inhibit lung adenocarcinoma A549 cells migration in concentration-depend manners (P < 0. 01). After treated with 25, 50,100 μg /ml RAGE ligands CML-AGE,the expression of anti-apoptotic gene BCL-2 was down-regulated and proapoptotic gene BAX was upregulated in the experimental group in concentration-depend manners(P < 0. 05),the difference was significant. Conclusion The expression levels of RAGE and DIAPH1 in lung adenocarcinoma A549 cells are both significantly lower than human bronchial epithelium cells BEAS-2B. RAGE ligands can inhibit cells migration and promote cell apoptosis in lung adenocarcinoma A549 cells and may provide a new target for the therapy of lung adenocarcinoma cells.

Utility of MOC-31 monoclonal antibody in differentiating metastatic adenocarcinoma cells and reactive mesothelial cells in effusion cytology

In effusion cytology, a clear distinction between reactive mesothelial cells and metastatic adenocarcinoma cells is sometimes challenging mainly due to similarities in the cytomorphological features. In such cases for definitive diagnosis, paraffin-embedded cell block examination and immunohistochemistry are helpful in making this distinction. MOC-31 is one of the proposed immunomarker for adenocarcinoma cells. We undertook to evaluate the role of MOC-31 as a marker for identifying adenocarcinoma cells in effusion specimen. A total of 185 paraffin-embedded cell blocks of effusion samples were identified, of these 111 cases were of metastatic adenocarcinoma. MOC-31 was positive in 101 of the 111 cases of metastatic adenocarcinoma. Minimal focal cytoplasmic staining was also seen in 7 of the 74 cases of reactive mesothelial cells, but these were taken negative as they did not show membrane positivity. The sensitivity and specificity of MOC-31 for metastatic adenocarcinoma cells were 92.5%, and 100% respectively, positive and negative predictive value (NPV) was 100% and 91.14%, respectively. MOC-31 can be used as a reliable marker in effusions for distinguishing metastatic adenocarcinoma from reactive mesothelial cases.

Combination effect of oncolytic virus H101 and radiation on apoptosis induction and toxicity in A549 lung adenocarcinoma cells / 中华放射医学与防护杂志

Objective To investigate the apoptosis and toxicity of oncolytic virus H101 combined with radiation on apoptosis of A549 lung adenocarcinoma cells. Methods A549 lung adenocarcinoma cells in exponential growth phase were divided into four groups: control ( PBS) group, radiation ( IR) group, oncolytic virus (H101) group and radiation combined with oncolytic virus (IR+H101) group. The cells were double dyed with Annexin fluorescein isothiocyanate ( V-FITC/PI ) and then the apoptosis ratio of cells in every group was detected by the flow cytometry. The cytotoxic effect of cells in every group was detected by lactate dehydrogenase ( LDH) release test. The mRNA expression of oncolytic viruses H101 capsid protein Hexon was detected by real-time fluorescence PCR ( RT-PCR) to compare the oncolytic virus replication in each group. Results Cell apoptosis rate in H101 group (55. 37%) was significantly higher than that in PBS group (1.03%) (t =36.51, P <0.05). Cell apoptosis rate in IR + H101 group (93. 06%) was significantly higher than that in H101 group (55. 37%), IR group (12. 67%) and PBS group (1. 03%) (t=13. 51, 24. 14, 38. 99, P<0. 05). LDH releasing percentage in IR group and H101 group at different time after virus transfection was significantly higher than that in PBS group ( t=25. 84,39. 38, 32. 51, 78. 18, P<0. 05;t=31. 40, 2. 68, 23. 43, 60. 98, P<0. 05). LDH releasing percentage in IR+H101 group was significantly higher than that in PBS group (t=80. 71, 119. 74, 109. 80, 123. 94, P<0. 05), IR group (t=28. 80, 54. 34, 72. 34, 61. 91, P<0. 05) and H101 group (t=42. 02, 57. 45, 57. 01, 58. 83, P<0. 05). Compared with H101 group at the same time point, the mRNA expression of Hexon in IR + H101 group at 24, 48 and 72 h was increased by 16. 26, 28. 37 and 39. 58 times, respectively (t=54. 50, 33. 73, 29. 28, P<0. 05). Conclusions The oncolytic virus H101 plays a role of radiosensitization in tumor cells. Radiation also increases the replication of the oncolytic virus H101 and thereby enhances the oncolytic effect of the oncolytic virus H101. Therefore, oncolytic virus H101 combined with radiotherapy has synergistic effect on killing tumor cells.

Combination effect of oncolytic virus H101 and radiation on apoptosis induction and toxicity in A549 lung adenocarcinoma cells / 中华放射医学与防护杂志

Objective To investigate the apoptosis and toxicity of oncolytic virus H101 combined with radiation on apoptosis of A549 lung adenocarcinoma cells. Methods A549 lung adenocarcinoma cells in exponential growth phase were divided into four groups: control ( PBS) group, radiation ( IR) group, oncolytic virus (H101) group and radiation combined with oncolytic virus (IR+H101) group. The cells were double dyed with Annexin fluorescein isothiocyanate ( V-FITC/PI ) and then the apoptosis ratio of cells in every group was detected by the flow cytometry. The cytotoxic effect of cells in every group was detected by lactate dehydrogenase ( LDH) release test. The mRNA expression of oncolytic viruses H101 capsid protein Hexon was detected by real-time fluorescence PCR ( RT-PCR) to compare the oncolytic virus replication in each group. Results Cell apoptosis rate in H101 group (55. 37%) was significantly higher than that in PBS group (1.03%) (t =36.51, P <0.05). Cell apoptosis rate in IR + H101 group (93. 06%) was significantly higher than that in H101 group (55. 37%), IR group (12. 67%) and PBS group (1. 03%) (t=13. 51, 24. 14, 38. 99, P<0. 05). LDH releasing percentage in IR group and H101 group at different time after virus transfection was significantly higher than that in PBS group ( t=25. 84,39. 38, 32. 51, 78. 18, P<0. 05;t=31. 40, 2. 68, 23. 43, 60. 98, P<0. 05). LDH releasing percentage in IR+H101 group was significantly higher than that in PBS group (t=80. 71, 119. 74, 109. 80, 123. 94, P<0. 05), IR group (t=28. 80, 54. 34, 72. 34, 61. 91, P<0. 05) and H101 group (t=42. 02, 57. 45, 57. 01, 58. 83, P<0. 05). Compared with H101 group at the same time point, the mRNA expression of Hexon in IR + H101 group at 24, 48 and 72 h was increased by 16. 26, 28. 37 and 39. 58 times, respectively (t=54. 50, 33. 73, 29. 28, P<0. 05). Conclusions The oncolytic virus H101 plays a role of radiosensitization in tumor cells. Radiation also increases the replication of the oncolytic virus H101 and thereby enhances the oncolytic effect of the oncolytic virus H101. Therefore, oncolytic virus H101 combined with radiotherapy has synergistic effect on killing tumor cells.

Effect of Livin on Proliferation and Multi-drug Resistance of Lung Cancer Cells A549 / 中国医科大学学报

Objective To study the effect of the inhibitor of apoptosis protein,Livin on proliferation and multi?drug resistance of lung adenocarcino?ma cells A549. Methods A549 cells were transfected with the eukaryotic expression vector pcDNA3.1?Livin. A549 cell clone with stable expres?sion of Livin was obtained through G418 screening. Expressions of Livin mRNA and protein in the transfected cells were respectively measured by re?verse transcription polymerase chain reaction(RT?PCR)and Western blot. The distribution of cell cycle phase was determined using flow cytometry. The level of P?gp mRNA and protein in A549 cells transfected with pcDNA3.1?Livin was detected by RT?PCR and Western blot. The analysis of multi?drug resistance of A549 treated with different chemotherapeutics was performed by MTT. Results The mRNA and protein expressions of Liv?in were both significantly increased in the transfected A549 cells. The flow cytometry analysis showed there was higher percentage of S phase and low?er percentage of G0/G1 phase in A549 cells transfected with pcDNA3.1?Livin. Compared with control groups,the expression of P?gp mRNA and pro?tein was increased in A549 cells transfected with pcDNA3.1?Livin,which showed a higher drug resistance and lower sensitivity to chemotherapic drugs such as ADM,MTX,CTX,and DDP(P<0.05). Conclusion Overexpression of Livin could enhance the proliferation of A549 cells,and high expression of P?gp caused by Livin could serve as one of the causes for multi?drug resistance in lung adenocarcinoma against chemotherapies.

Effect of C-erbB-2 shRNA on chemosensitivity of mouse lung adenocarcinoma cells and its mechanism / 吉林大学学报(医学版)

Objective To investigate the effect of C-erbB-2 shRNA on chemosensitivity of mouse lung adenocarcinoma cells and its mechanism,and to find new therapy method for non-small cell lung cancer,especial lung adenocarcinoma.Methods The mouse lung adenocarcinoma Lewis cells were cultured regularly and divided into non-transfected group, pGPU6/RFP/Neo-shNC group and pGPU6/RFP/Neo-erbB-2 group. The plasmids were synthesized and transfected into Lewis cells in each group by Lipofectamine 2000.The expression levels of C-erbB-2 mRNA and protein in the cells in various groups were tested by RT-PCR and Western blotting method, respectively. The Lewis cells were divided into non- transfected group, pGPU6/RFP/Neo-shNC group, carboplatin group, pGPU6/RFP/Neo-erbB-2 group, pGPU6/RFP/Neo-shNC+carboplatin group and pGPU6/RFP/Neo-erbB-2+ carboplatin group. The apoptotic rates of the cells in each group were detected by flow cytometry;the expression levels of Bcl-2 and Bax proteins in each group were determined by Western blotting method.Results The expression levels of C-erbB-2 mRNA and protein in pGPU6/RFP/Neo-erbB-2 group were lower than those in non-transfected group and pGPU6/RFP/Neo-shNC.The apoptotic rate of the cells in pGPU6/RFP/Neo-erbB-2+carboplatin group was the highest in all of the groups (P<0.01);compared with the others, the expression of Bax protein in pGPU6/RFP/Neo-erbB-2+carboplatin group was increased, while the expression level of Bcl-2 protein was decreased.Conclusion C-erbB-2 shRNA can increase the Lewis cells’sensitivity to carboplatin.The mechanism may be that it can enhance the Lewis cells’apoptosis induced by carboplatin through increasing the expression of Bax protein and decreasing the expression of Bcl-2 protein.

Effect of rapamycin on proliferation, invasion, adhesion, apoptosis and autophagy of human lung adenocarcinoma A549 and resistant A549/DDP cells treated with cis-diamminedichloroplatinum / 中国病理生理杂志

[ ABSTRACT] AIM:To study the effect of rapamycin ( Rap) on the proliferation, invasion, adhesion, apoptosis and autophagy of human adenocarcinoma A549 and resistant A549/DDP cells treated with cis-diamminedichloroplatinum ( DDP) .METHODS: Human adenocarcinoma A549 and resistant A549/DDP cell lines were cultured.The inhibitory effects of Rap alone or combined with DDP on A549 and resistant A549/DDP cells were detected by MTT assay.The in vitro invasion abilities of the 2 cell lines treated with Rap alone or combined with DDP were detected by Transwell methods. The in vitro adhesion abilities of the 2 cell lines treated with Rap alone or combined with DDP were detected by adhesion experiments.The apoptosis of A549 and resistant A549/DDP cells induced by Rap alone or combined with DDP was ana-lyzed by flow cytometry.The cell autophagy marker proteins beclin-1 and LC3 in A549 and resistant A549/DDP cells trea-ted with Rap alone or combined with DDP were detected by Western blotting.RESULTS:Compared with Rap or DDP a-lone group, the combination of Rap and DDP significantly inhibited the proliferation, invasion and adhesion of A549 and resistant A549/DDP cells in vitro, and promoted the cell apoptosis and autophagy marker proteins beclin-1 and LC3 expres-sion ( all P<0.05) .CONCLUSION:Rap enhances the effect of DDP through promoting the cell autophagy, thereby in-hibiting the proliferation, invasion and adhesion of A549 and resistant A549/DDP cells and inducing the cell apoptosis with a synergistic effect.

Pharmacokinetic aspects and in vitro-in vivo correlation potential for lipid-based formulations

Lipid-based formulations have been an attractive choice among novel drug delivery systems for enhancing the solubility and bioavailability of poorly soluble drugs due to their ability to keep the drug in solubilized state in the gastrointestinal tract. These formulations offer multiple advantages such as reduction in food effect and inter-individual variability, ease of preparation, and the possibility of manufacturing using common excipients available in the market. Despite these advantages, very few products are available in the present market, perhaps due to limited knowledge in the in vitro tests (for prediction of in vivo fate) and lack of understanding of the mechanisms behind pharmacokinetic and biopharmaceutical aspects of lipid formulations after oral administration. The current review aims to provide a detailed understanding of the in vivo processing steps involved after oral administration of lipid formulations, their pharmacokinetic aspects and in vitro in vivo correlation (IVIVC) perspectives. Various pharmacokinetic and biopharmaceutical aspects such as formulation dispersion and lipid digestion, bioavailability enhancement mechanisms, impact of excipients on efflux transporters, and lymphatic transport are discussed with examples. In addition, various IVIVC approaches towards predicting in vivo data from in vitro dispersion/precipitation, in vitro lipolysis and ex vivo permeation studies are also discussed in detail with help of case studies.

Soybean extracts enriched with free isoflavones promote nitric oxide synthesis and affect the proliferation of breast adenocarcinoma cells

Although soybean isoflavones naturally accumulate in their conjugated forms, the beneficial effects on human health of soybean-containing foods have been credited to their aglycone forms. In the present study we analyzed the effects of a chemical agent, sodium nitroprusside (SNP), in eliciting the exudation of non-conjugated isoflavones from intact soybean seeds, embrionary axes and cotyledons. The isoflavones in the exudates were determined by high performance liquid chromatography and mass spectrometry. The effect of the exudates on the emission of nitric oxide (NO) and on the proliferation of breast carcinoma cells (MCF-7) was also evaluated. SNP elicitation increased the production of the aglycone forms dose- and time-dependently. Exudates of embrionary axes and cotyledons stimulated NO emission and showed biphasic effects on viability of MCF-7 cells. At lower concentrations both extracts presented proliferative effects (10-25%), and at higher concentrations inhibited (15%) cell proliferation. The biphasic effect might be due to the action of isoflavone aglycones in activating estrogen receptors which in turn stimulate the production of NO. Overall, the results suggest that soybean extracts enriched in isoflavone aglycones by elicitation with SNP could be exploited as a functional ingredient in the food industry.

The cytotoxic and growth inhibitory effects of palladium(II) complexes on MDA-MB-435 cells

The antitumorigenic potential of two palladium(II) complexes, [Pd(ca2-o-phen)Cl2 ] - C1 and [Pd(dmba)(dppp)Cl] - C2, was evaluated, using MDA-MB-435 cells, a human breast adenocarcinoma cell-line that does not express the estrogen receptor α (ER-). Growth inhibition and induced alterations in cell-morphology were analyzed. The sulforhodamine B test showed that, compared to control cells, both C1 and C2 significantly inhibited (p < 0.5) cell growth. The maximum effect with both was achieved with 1 µM complexes, after 24 h of treatment. No further cell-growth inhibition was achieved by increasing concentration or incubation time. Cell morphology was analyzed after staining with hematoxylin-eosin (HE). The morphological changes noted in the treated cells were cell rounding-up, shrinkage, nuclear condensation and reduction of cell length (p < 0.05), thereby indicating that both C1 and C2 are cytotoxic to breast adenocarcinoma cells. All together, there was every indication that, by decreasing cell growth and inducing morphological changes, the tested complexes are cytotoxic, hence their potentiality as promising candidates for antineoplastic drug development.

Vascular endothelial growth factor receptor targeted RNA interference inhibits growth of human lung adenocarcinoma cells / 第二军医大学学报

Objective: To investigate the inhibitory effects of RNA silencing via adenovirus-mediated vascular endothelial growth factor receptor (VEGFR) shRNA on proliferation of lung adenocarcinoma cells in vitro and in vivo. Methods: Ad-VEGFRshRNA adenovirus containing enhanced green fluorescent protein (EGFP) gene and VEGFRshRNA was constructed and was used to infect A549 cells; fluorescent microscopy was used to observe the infection efficiency. Western blotting assay was used to examine the expression of VEGFR protein in A549 cells. MTT method was used to examine the cell viability and the cell growth curve was drawn. The inhibition of cell growth was examined by cell cycle and colony-forming test. Meanwhile, nude mice were transplanted with A549 cells to establish tumor-bearing model, and the long term growth of tumor was observed. Results: Western blotting revealed that the expression of VEGFR was obviously decreased in the RNA interference group. The cell growth curve indicated that the cell growth was obviously inhibited after RNA interference. Cell cycle and colony-forming test indicated that the tumor growth was obviously inhibited after RNA interference. In vivo study with nude mice also indicated that RNA interference obviously inhibited tumor growth. Conclusion: The constructed VEGFR-targeted shRNA can effectively inhibit VEGFR expression in A549 cells and can suppress the growth of A549 cells.

Transfection of T-STAR antisense gene can depress the telomerase activity in lung adenocarcinoma cell line A549 / 第三军医大学学报

Objective To investigate the changes of telomerase activity after knocking down endogenous expression of T-STAR (testes-signal transduction and activator of RNA) gene in human lung adenocarcinoma cell line A549 by antisense strategy. Methods The mRNA and protein expression of T-STAR gene were determined by RT-PCR and Western blotting, and the telomerase activity was measured by PCR-ELISA, after transfection of T-STAR antisense gene into A549 cells with lipofectamine. Sense pcDNA-STAR and blank pcDNA3.1 transfection served as control. Results The expression of T-STAR gene was significantly inhibited at mRNA and protein level, and the telomerase activity was significantly decreased. Conclusion The down-regulation of telomerase activity may result from inhibition of T-STAR gene expression in lung adenocarcinoma A549 cells.

Effects ofcis-9,trans-11-conjugated linoleic acid on cancer cell cycle

<p><b>OBJECTIVES</b>To determine the effect of cis-9, trans-11-conjugated linoleic acid on the cell cycle of mammary cancer cells (MCF-7) and its possible mechanism of inhibition cancer growth.</p><p><b>METHODS</b>Using cell culture and immunocytochemical techniques, we examined the cell growth, DNA synthesis, expression of PCNA, cyclin A, B(1), D(1), p16(ink4a) and p21(cip/wafl) of MCF-7 cells which were treated with various c9, t11-CLA concentrations (25 mM, 50 mM, 100 mM and 200 mM) of c9, t11-CLA for 24 and 48 h, with negative controls (0.1% ethanol).</p><p><b>RESULTS</b>The cell growth and DNA synthesis of MCF-7 cells were inhibited by c9, t11-CLA. MCF-7 cells, after treatment with various c9, t11-CLA doses mentioned above for 8 days, the inhibition frequency was 27.18%, 35.43%, 91.05%, and 92.86%, respectively and the inhibitory effect of c9, t11-CLA on DNA synthesis (except for 25 mM, 24 h) incorporated significantly less(3)H-TdR than did the negative control (P<0.05 andP<0.01). To further investigate the influence on the cell cycle progression, we found that c9, t11-CLA may arrest the cell cycle of MCF-7 cells. Immunocytochemical staining demonstrated that MCF-7 cells preincubated in media supplemented with different c9, t11-CLA concentrations at various times significantly decreased the expressions of PCNA, and Cyclin, A, B(1), D(1) compared with the negative controls (P<0.01), whereas the expressions of p16(ink4a) and p21(cip/wafl), cyclin-dependent kinases inhibitors (CDKI), were increased.</p><p><b>CONCLUSIONS</b>The cell growth and proliferation of MCF-7 cells is inhibited by c9, t11-CLA by blocking the cell cycle, which reduces expressions of cyclin A, B(1), D(1) and enhances expressions of CDKI (p16(ink4a) and p21(cip/wafl)).</p>

The Inhibitory Effect of Amiloride on the Growth of Human Gastric Carcinoma Cells in Vitro / 대한암학회지

PURPOSE: In the present study the effects of amiloride on the growth of human gastric adenocarcinoma cell line, AGS cells were examined with or without the addition of 5-fluorouracil (5-FU) in vitro. MATERIALS AND METHODS: The growth of AGS cells was examined by counting number of cells on two and four days post-treatment with 50 micrometer, 100 micrometer, 200 micrometer, 400 micrometer, 800 micrometer, amiloride, and 0.1 microgram/ml, 0.3 microgram/ml 5-FU, after plating AGS cells into 6 well plates at a density of 10 x 10(4) cells/well. The reversibility of the effects of amiloride was examined on two to eight days post-treatment with 400 micrometer amiloride after seeding 2 x 10(4) cells/dish. Cell cycle analysis was performed after four day-treatment with 400 micrometer amiloride. RESULTS: Amiloride (50~800 micrometer) significantly inhibited the growth of AGS in a dose-dependent fashion (p<0.05). The inhibitory effect of amiloride on growth of AGS was reversible since removal of amiloride after 24 hours treatment led to resumption of rapid growth up to control levels. Amiloride combined with 5-FU markedly inhibited the growth of AGS in a dose-dependent fashion compared to that of amiloride or 5-FU alone (p<0.05). The fraction of S phase, G0-G1 phase and G2-M phase was 19.3%, 55.7%, 18.8%, in the amioride group (400 micrometer) and 43.9%, 37.4%, 25.1% in the control group, respectively, showing significantly higher G1 fraction in amiloride group compared to control. CONCLUSION: This is the first paper which reported that amiloride inhibited in vitro growth of human gastric adenocarcinoma cells and that its effect of growth inhibition may be synergistic with 5-FU. Amiloride given with or without 5-FU may be useful agent in the treatment of gastric carcinomas. The inhibitory effects of amiloride on the growth of AGS may be mediated in part by blocking G1-S transition of cell cycle.

Effects of Qingjin Desheng Tablets on Telomerase and Protein Expressions of Apoptosis Relevant Genes in Human Lung Adenocarcinoma Cells / 中药新药与临床药理

Objective To explore the molecular mechanism of Qingjin Desheng Tablets on inhibiting the growth of lung cancer cells and to supply experimental evidences for Chinese herbal medicine preventing and curing lung cancer in clinical application .Methods Immunohistochemistry staining was used to detect the activity of telomerase and the expression of bcl-2 and bax in human lung adenocarcinoma cells after treated with serum containing Qingjin Desheng Tablets.Results The activity of telomerase was inhibited significantly and the positive rate of bcl-2 protein reduced,and the positive rate of bax2 increased in human lung adenocarcinoma cells after 72 hour-treatment with the serum containing Qingjin Desheng Tablets.Conlusion Qingjin Desheng Tablets exert the anti-tumor action through inhibiting the activity of telomerace and modulating the expression of bcl-2 and bax protein.

Influences of shRNA on survivin gene expression and sensitivity of A549 cells to paclitaxel / 中国药理学通报

Aim To construct the eukaryotic expression vectors of short hairpin RNA targeting survivin and observe its effect on biologic behavior of A549 cells and sensitivity of A549 cells to paclitaxel.Methods The DNA fragment targeting human survivin was inserted into the plasmid,and the recombinant plasmid was constructed.The recombinant plasmids cells were transfected into A549 cells by FuGENE transfection reagent.The expression levels of survivin gene were detected with RT-PCR and Western blot before and after transfection,respectively.Cell apoptosis was detected by TUNEL method.The sensitivity of A549 cells to paclitaxel was detected by MTT after transfection.Results The recombinant plasmid was successfully constructed.RNAi group cells showed lower expression of survivin than control group.The apoptosis rate of A549 cells increased after transfection.The IC50 of paclitaxel inhibiting A549 cells was 11.9 fold before transfection compared with those after transfection.There was significant difference between the two groups(P

Sensitization effect of insulin of chemotherapy on human adenocarcinoma cells A549 / 解放军医学杂志

Objective To explore the sensitization effect of insulin to chemotherapy as a metabolic promoter on human lung adenocarcinoma cells A549.Methods The sensitivity of human adenocarcinoma cells A549 to DDP was observed by MTT methods,and IC30 values of the cells were calculated.The optimal concentration of insulin to promoting A549 cells' growth was determined by MTT assay.The A549 cells were pretreated with insulin of various concentrations for different time period,and then the cell sensitivity to DDP was evaluated with MTT assay.Flow cytometry was used to study the cell cycle distribution of the A549 cells treated with insulin.Results The IC30 value of DDP was about 36.87?g/ml.It was found that the cell growth was significantly promoted with pretreatment of insulin at the concentration of 4~16mU/ml for 8~16 hours.Insulin enhanced the inhibitory effect of DDP(36.87?g/ml)on cell proliferation when human lung adenocarcinoma cells A549 were treated with insulin(2.0~16.0mU/ml;8~16h)as indicated by MTT assay(P