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OBJECTIVE： To conduct structural modification of tectorigenin to search for new compounds with anti-tumor activity. METHODS： Tectorigenin was used as a lead compound， and then added into amine reagents as ethanolamine， methylamine， ethylamine， dimethylamine， diethylamine， n-propylamine and formaldehyde solution. Tectorigenin Mannich base derivatives were synthesized by mannich reaction with as the lead compound. The structures of the derivatives were identified according to IR， UV， MS and NMR data. Solubility of tectorigenin and its derivatives were investigated by solubility test method. MTT assay was used to investigate the inhibitory effects of tectorigenin and its derivatives on the proliferation of human colon cancer cell line HCT116， human lung cancer cell line A549 and human hepatoma cell line HepG2， and half inhibitory concentration （IC50） was calculated. The inhibition rate of tectorigenin and its derivatives （100 mg/kg） on H22 hepatoma-bearing mice in vivo was studied. RESULTS： Totally of 6 kinds of tectorigenin mannich base derivatives were synthesized， such as 8-（N-hydroxyethyl）-methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N-methyl）-methyleneamino-5，7，4′-trihydroxy-6- methoxyisoflavone， 8-（N， N-diethyl）-methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N， N-dimethyl）-methyleneamino- 5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N-ethyl）-methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N-propyl）- methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone （compounds 1-6 in turn）. Compared with tectorigenin， the water solubility of six derivatives was significantly improved， and the solubility was 5-20 times higher than that of tectorigenin. IC50 of compounds 1， 3 and 5 to HCT116 cells were （34.82±3.27）， （16.21±4.13）， （33.12±3.25） μmol/L， which were stronger than that of tectorigenin [（45.23±5.74） μmol/L]； IC50 of compounds 1， 3 and 5 to A549 cells were （37.05±5.74）， （26.88±4.52）， （30.13±6.23） μmol/L， which were stronger than that of tectorigenin [（53.24±6.34） μmol/L]； IC50 of compounds 1， 3 and 5 to HepG2 cells were （23.74±1.45）， （18.96±2.34）， （30.95±2.87） μmol/L， which were stronger than that of tectorigenin [（48.98±2.58） μmol/L]. Compounds 1， 3 and 5 showed higher inhibition rates （55.51％， 57.20％ and 49.15％） than tectorigenin （33.05％） on H22 hepatoma-bearing mice， respectively. The other three compounds had no obvious advantage over tectorigenin in anti-tumor activity. CONCLUSIONS： In this study， compounds 1， 3 and 5 of six tectorigenin mannich base derivatives synthesized in this study have stronger antitumor activity than tectorigenin.
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Objective:To explore the effects ofbufalin (BUF) combined with doxorubicin (DOX) on the proliferation and apoptosis in human lung cancer cell line A549 in vitro.Methods:Methyl thiazolyl tetrazolium (MTT) assay was used to measure the inhibitory effects of BUF,DOX and their combination on the growth ofA549 cells.Hoechst 33342 staining was used to observe the changes of nucleus.Flow cytometry was used to investigate the apoptosis and cell cycle distribution of A549 cells.Western blot was used to examine the expression of apoptotic protein.Results:BUF and DOX showed inhibitory effect on the A549 cells in a dose and time-dependent manner.Compared with BUF or DOX alone,combination of BUF (1,20,100 nmol/L) with DOX (1.0 μg/mL) could significantly increase the growth inhibition rate ofA549 cells at 24,36,72 h,respectively (all P＜0.05).BUF and DOX alone could induce apoptosis,and their combination could significantly increase the apoptosis ratio.In addition,BUF combined with DOX could block the cell stage of A549 cells,keep the cell stage stay in S stage and up-regulate the expression of caspase-3.Conclusion:BUF combined with DOX can significantly inhibit the proliferation ofA549 cells,which might be related to the induction of apoptosis,cell cycle S phase arrest and caspase-3 up-regulation.
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Objective To investigate the effects of 4.1N expression in lung cancer A549 cell line on cell proliferation, invasion and migration. Methods A549 cells were cultured in vitro and transfected with lipofectamine 2000 mediation. Three groups were employed: transfection with pEGFP-4.1N plasmid, pEGFP vector plasmid, and blank control, respectively. The mRNA and protein expression differences of 4.1N was examined by semi-quantitative RT-PCR and Western blot in every group after 48 h. The proliferation capability was determined by MTT assay. Invasion capability was evaluated by scratches, adhesion experiments and Transwell chamber model. Results After the transfection, the expression of 4.1N mRNA and protein in pEGFP-4.1N plasmid transfection group was significantly enhanced (P<0.05). The proliferation capability of A549 cells descended extremely (P<0.05). The migration and invasion capability of A549 cells in vitro decreased substantially (P<0.05). Conclusions Transfected with 4.1N gene can significantly increases the expression levels of 4.1N mRNA and protein in A549 cells which are highly metastatic in human. Cell behavior in vitro studies showed that 4.1N gene can inhibit the proliferation, adhesion, invasion and migration of A549 cells, which plays an important role in the metastasis of lung cancer and it may become a molecular marker for metastasis of lung cancer.
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Objective: To investigate the effect of astragaloside IV on tumor cellular uptake and antitumor efficacy by ginsenoside compound K (GCK). Methods: The human lung adenocarcinoma cell line A549 was prepared as an antitumor model, the cytotoxicity of the mixtures of GCK and astragaloside IV to A549 cells was determined by MTT assay, the cellular uptake of GCK was detected by fluorescence microscopy, and the intracellular GCK was determined by HPLC. The enhancement of astragaloside IV in vivo efficacy by GCK was evaluated by nude mice bearing tumor model. Results: The effect of GCK on the inhibition of A549 cell proliferation was enhanced in the presence of astragaloside IV and astragaloside IV could increase the uptake rate of GCK in A549 cells, with the proportion of astragaloside IV increasing, the cytotoxicity was significantly stronger than GCK and the uptake rate was improved. In vivo antitumor assay of mice bearing tumor indicated that astragaloside IV could enhance the antitumor efficacy of GCK. Conclusion: Preliminary results indicate that the mixture of GCK and astragaloside IV have the effect of enhancing antitumor efficacy.
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Aim To investigate the anticancer effect of a new xanthono-pyridine derivative N, N '-( 7-oxo-7H-chromeno[3,2-h] quinoline-5,9-diyl)-bis(2-( pyrroli-din-1-yl)acetamide) (XP-16) on human lung carcino-ma cell line A549 and the potential mechanism. Meth-ods Antiproliferative effect of XP-16 on A549 cells was evaluated by MTT assay, morphological examina-tion and colonial assay. Apoptosis detection was car-ried out using Hoechst 33258 and PI double-dyeing method. Intracellular Ca2+ concentration ( [ Ca2+] i ) and mitochondria membrane potential were detected by fluorospectrophotometer. A549 cells treated with XP-16 were collected for Bad and metallothionein 1 A ( MT-1 A ) transcript analysis by real-time reverse tran-scriptase-polymerase chain reaction ( qRT-PCR) . Re-sults XP-16 inhibited A549 cell proliferation in dose-and time-dependent manner. Typical apoptotic mor- phology such as chromatin aggregation and nuclear fragmentation was observed in A549 cells treated with XP-16 for 24 h, and the apoptosis was showed in a dose-dependent manner. After treated with XP-16, [ Ca2+] i and mitochondria membrane potential of A549 cells were decreased, and relative mRNA level of Bad and MT-1A was up-regulated. Conclusions XP-16 has anticancer effect on A549 cells through apoptosis, which might be associated with decreasing intracellular Ca2+ concentration and mitochondria membrane poten-tial. Up-regulation of MT-1A expression might be the result of decreased [ Ca2+] i .
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Objective:To observe the effects of genistein on proliferation and apoptosis of human non -small cell lung cancer cell line A549/DDP.Methods:①MTT assay was applied to evaluate the resistance index of A 549/DDP cell line to cisplatin and half in-hibitory concentration ( IC50 ) .②Inhibition rate of A549/DDP cell proliferation and IC 50 value were evaluated by MTT assay after treat-ment with 0, 1.25, 2.5, 5.0, 10, 20, 40, 60, 80 μg/ml genistein for 48 hour respectively.③A549/DDP cell cycle and apoptosis were evaluated by flow cytometry after treatment with 6.25, 12.5, 25 μg/ml genistein for 24 hours respectively.Results:①In expo-sing to cisplatin, the IC50 of A549 and A549/DDP was 33.6 μmol/L and 76.9 μmol/L respectively.The resistance index was 2.3. Cell growth inhibition rate increased following the cisplatin concentration increasing gradually .②A549/DDP growth inhibition rate in-creased at first and later decreased gradually following treatment with the genistein dose increased .The IC50 of A549 and A549/DDP was about 85.1 μg/ml and 80.2μg/ml respectively.③After treatment with 6.25, 12.5, 25μg/ml genistein for 24 hours, there were more A549/DDP cells arresting and showing apoptosis along with the genistein dose increased .Conclusion: Genistein can inhibit A549/DDP proliferation, cause A549/DDP arresting in G2/M phase and induce A549/DDP cell apoptosis with dose dependently .
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In order to investigate the inhibitory effects of Endostar(rh-endostatin,YH-16)in combination with radiotherapy on lung adenocarcinoma A549 in mice and the interaction mechanisms of combined therapy,the transplantation tumor models of A549 lung adcnocarcinoma were established.When the largest diameter of tumor reached 1.0 cm,all nude mice were randomly divided into 4 groups: Endostar group,radiotherapy group,radiotherapy plus Endostar(combined treatment)group,and control group(n=6 in each group).The largest diameter and the vertical diameter of tumor were measured at different time points.At the 16th day,mice were executed,and the tumors were applied to analysis of rate of tumor cell apoptosis,and the expression levels of basic fibroblast growth factor(bFGF)mRNA were detected by reverse transcription-polymerase chain reaction(RT-PCR)and those of vascular endothelial growth factor(VEGF)by immunohistochemistry.The results demonstrated that the rate of tumor inhibition in combined treatment group was higher than that in other groups.And the rate of tumor cell apoptosis in combined treatment group was also higher than that in other groups.Meanwhile,the levels of bFGF mRNA and VEGF expression in combined treatment group were lower than those in other groups.It was concluded that Endostar obviously enhanced the curative effectiveness of radiotherapy on lung adenocarcinoma A549 in mice.The underlying mechanisms may involve the down-regulation of bFGF mRNA and VEGF expression to inhibit angiogenesis by Endostar and the cooperative effect of Endostar and radiotherapy to synergistically promote tumor cell apoptosis.And Endostar inhibits angiogenesis by down-regulating the expression of bFGF mRNA and VEGF.
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Objective:Toinvestigate the influence of MMP7 antisense oligodeoxynucleotide(ASODN)on adhesion and invasion ofhuman lung adenocarcinoma cell line A549.Methods:Phosphorothioate MMP7 ASODN was transfected to A549 cells mediated by liposome. The expression of MMP7 was examined by RT-PCR.The adhesive and invasive ability were examined by plate adhesion model and Boyden Chamber transwell assay.Results:After MMP7 ASODN was transfected,the relative optical density(ROD)of electrophoresis strip was decreased obviously(P
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Objective To observe the effect of Hydroxycamptothecin (HCPT) on proliferation of human lung cancer cell line A549 in vitro. Method Using cell calture and MTT assay to observe the effect of HCPT on proliferation of human lung cancer cell line A549. Result Lower concentration of HCPT had no evident inhibitory effect on the human lung cancer cell line A549 after 24 h. The effect was evident when the concentration of HCPT was up to 50 ?g/mL, and the inhibitory rate was 44.17%. The inhibitory rate was 50.28% when the concentration of HCPT was up to 100 ?g/mL. The inhibitory effect of HCPT became more significant with the stimulation of the time, and the inhibitory rate of 100 ?g/mL concentration of HCPT was 70.98% after 48 h. Conclusions HCPT can inhibit the proliferation of human lung cancer cell A549 in vitro. The effect is dose and time dependent.
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OBJECTIVE:To investigate the effect of liposome-encapsulated demethoxycurcumin on the proliferation of human lung adenocarcinoma cell line A549.METHODS:The in vitro cultured human lung adenocarcinoma cell line A549 was treated with liposome-encapsulated Bisdemethoxycurcumin.The bionomics including morphology and cell cycle of cell line A549 were observed by morphological analysis,MTT assay and flow cytometry assay.RESULTS:Marked morphological changes were noted in cell line A549 under microscope.Liposome-encapsulated Bisdemethoxycurcumin showed inhibitory action on the growth of A549 cell lines,with IC50 at 12.108?g/mL.The progression of cell cycle was arrested in S phase.CONCLUSION:Liposome-encapsulated Bisdemethoxycurcumin can inhibit the growth of human lung adenocarcinoma cell line A549,which has a promising future in the treatment of adenocarcinoma of lung.
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Objective To study the effect of the recombinant matrilysin (rMMP-7) and its antisense oligonucleotide transfected by liposome (LIPO-MAON) on the proliferation of lung adenocarcinoma cell line A549 and human umbilical vein endothelial cells (HUVEC). Methods Antisense oligonucleotide of matrilysin was constructed by liposome transfection. The proliferation of HUVEC and A549 was detected by MTT assay in case of rMMP-7 or LIPO-MAON existence. The expression of MMP-7 mRNA in both HUVEC and A549 was examined by semi-quantitative RT-PCR assay. Results The proliferation of both HUVEC and A549 cell line was accelerated by high level of rMMP-7 (0.75, 1.0 ?g/ml), and the inhibited proliferation was only found in A549 cell line treated with high concentration of LIPO-MAON (7.5, 10 nmol/ml), but not in HUVEC. By RT-PCR assay, no expression of MMP-7 mRNA was found in HUVEC; however, enhanced or decreased expression of mRNA were found when A549 cell line was treated respectively with rMMP-7 or LIPO-MAON (P
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Objective To investigate the possibility of inhibiting the malignant proliferation of tumor cells in vitro through down-regulating the expression of MMP-7 by antisense gene technology. Methods A Phosphorothioate antisense oligodeoxynucleotide (ASODN) and scrambled oligodeoxynucleotide (SCODN) against MMP-7 were respectively transfected into A549 cells mediated by liposome. The expression of MMP-7 and cell proliferation in the cells were respectively examined by immunohistochemical assay and MTT. Flow cytometry was used to detect the cell cycles. Results After A549 cells were transfected with MMP-7 ASODN, the cell number was decreased. MTT method indicated that the proliferation of A549 cells was inhibited by MMP-7 ASODN at different concentrations, with the inhibition reaching the peak at 48 h. ASODN transfection downregulated the expression levels of MMP-7 protein, and inhibited the transition period of G_ 0 /G_ 1 phase to S phase. Conclusion The artificial MMP-7 ASODN can specifically inhibit the expression of MMP-7 protein and regulate cell cycle and cell proliferation in A549 cells.