Effect of Thalidomide on the Expressions of Vascular Endothelial Growth Factor in Human Lung Adenocarcinoma Cell Line
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To explore the effects of thalidomide on the expressions of vasc ular growth factors in parental and cisplatin_resistant human lung adenocarcinom a cell line A549 and A549 .Methods:RT_PCR and Immunohistochemistry were used to detect mRNA and protein expression of VEGF in A549 and A549 .Results:From t he 1st to 5 th day after treatment with physiological concentration of thalidomi de(6 mg/L),VEGF mRNA expression levels in A549 were significant higher than that before treatment.VEGF mRNA expression levels in A549 were significant lower. There were significant differences in A549 and A549 among different concentra tion of thalidomide on the 5 th day.The protein expressions were significant coi ncided with the relative VEGF mRNA expression levels in A549 and A549 .Conclus ion:Physiological concentration of thalidomide up_regulated VEGF mRNA expression significantly in A549.But large dose of thalidomide inhibits VEGF expression si gnificantly.Moreover,thalidomide down_regulated VEGF mRNA expression significant ly in A549 .It is dose_independent.The protein expression is significantly coi ncided with the relative VEGF mRNA.Cite
Aim To observe the regulatory effect of LZBZY on the growth of H22 tumor and the expression of vascular endothelial growth factor(VEGF) in transplanted H22 tumor of mice,and explore whether the inhibition of tumor is related to the inhibition of vascular growth.Methods The mice were vaccinated with H22 cells suspension(1×107/ml) 0.2 ml each one.The observation was made on the weight of the transplanted tumor of the mice,then the inhibitory rate was calculated.The expression of VEGF was determined by the immunohistochemical method.Results The LZBZY group had significant effect on inhibiting the tumor growth(P0.05,P0.01),when compared with the model group,the pathological detective results showed that the positive rate of expression of VEGF was 100% in each group with the degree difference.Moreover,the expression of VEGF was decreased obviously in LZBZY group(P0.05,P0.01).Conclusion The LZBZY can inhibit the growth of the transplanted H22 tumor of mice and decrease the expression of the VEGF.
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Objective To study the inhibitory effects of Protopanaxidiol(Ppd) on vascular endothelial growth factor(VEGF) and its mRNA expression of liver cancer.Methods Liver cancer model animals were divided into control group,positive drug group,cyclophosphamide group,low,medium and high dosage of Ppd group(25,50,100 mg/kg),10 ones in each group.After two weeks,the animals were killed to make histological section for immunohistochemical and hybridization in situ stain.Results Control group had increased tumor interstitial vessel density,the enhanced VEGF and mRNA expression positively relating to vessel density,while Ppd treatment group had decreased above indexes(P0.01);and medium and high dosage of Ppd had stronger inhibitory effect than cyclophosphamide(P0.01).Conclusions Ppd inhibits VEGF and its mRNA expression in tumor tissue and tumor angiogenesis,thus inhibits tumor growth.
Liver Cancer
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To study the effects of Rapamycin (RPM) on angiogenesis and tumor progression in human hepatocellular carcinoma (HCC) implantation mice.Tumor tissues of HCC were implanted into the liver of nude mice. Then, nude mice were treated with RPM and cyclosporine A (CsA). Real-time PCR was used to detect the mRNA expression of vascular endothelial growth factor (VEGF). Immunohistochemical stain and image analysis were used to detect the protein expression of VEGF and proliferating cell nuclear antigen (PCNA) and microvessel density (MVD) was counted by endothelial cells immunostained by anti-CD34 antibody. The concentration of VEGF in the peripheral blood was detected by ELISA.(1) The tumor weights were (372 +/- 35) mg, (769 +/- 39) mg and (751 +/- 42) mg in RPM, CsA and control group respectively. The tumor weight was significantly decreased in RPM group and no difference in CsA group compared with control group. (2) The expression of VEGF mRNA, VEGF and PCNA protein in tumor tissues and concentration of VEGF in the peripheral blood were remarkably down-regulated in RPM group compared with control group (P < 0.05) and were not remarkably different in CsA group from in control (P > 0.05).(3) Comparing with the control, the tumor MVD was remarkably decreased in RPM group (P < 0.05), and no difference in CsA group (P > 0.05).RPM can inhibit angiogenesis and tumor progression of HCC by down-regulated the gene and protein expression of VEGF and inhibited the growth of tumor.
Tumor progression
CD31
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Objective To study the effect of inhibitor of cyclooxygenase-2 on the expression of vascular endothelial growth factor(VEGF)in A549 cell lines of non-sman cell lung cancers. Methods The proliferation of A549 lung carcinoma cells was measured by MTT assay After A549 cells were treated with different dose celecoxib; After A549 cells were treated with celecoxib 50μmol/L for 48 h, the level of VEGF m RNA was detected RT-PCR, and the expressions of VEGF protein was detected by western blot method. Results Celecoxib could inhibit the proliferation of A549 cell by a time-and dose-dependent manner. Compared with control group, the level of VEGF m RNA was decreased significantly detected by RT-PCR after A549 cells were trealed with celecoxib 50μmol·L-1for 48 h, P0.01 and the expression of VEGF protein was decreased significantly detected by western blot method, P0.01.Conclusion celecoxib might inhibite the proliferation of A549 lung carcinoma cells by regulating the expressions of VEGF.
Celecoxib
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To observe the effect of drug-containing serum of Chinese traditional herbal decoction Weikangning (WKN) on growth of gastric cancer cell, expression of vascular endothelial growth factor (VEGF) and its receptors including KDR and Flt-1.A total of 120 male Wistar rats were given high, medium and low-dose WKN. After the drug-containing serum was prepared, the gastric cancer cells MGC-803 of different dose groups were cultured with the drug-containing serum, respectively. The gastric cell growth was observed by using light microscope and flow cytometer,the expression of VEGF and its receptor Flt-1 was detected with SABC immunohistochemistry method and the mRNA expression levels of VEGF and its receptors including KDR and Flt-1 of different groups were detected with reverse transcription-polymerase chain reaction (RT-PCR), respectively.The gastric cancer cell growth and cell cycle of the three medicated groups were significantly improved as compared with those of the control group (P <0. 01) ,the proportion of cells in G0 - G1 phase was increased,while the cells in S phase were decreased. It was shown that the apoptotic rates were increased in the medicated groups in a dose-dependent manner. The gray scales ( microm(2) ) of VEGF and Flt-1 in high, medium and low dose groups was 182. 44 +/-0. 54,178. 65 +/-0. 56,174. 80 +/-0. 81 and 168. 51 +/- 0. 81,162. 01 +/-0. 52,148. 20 +/-0. 69, respectively vs 147.82 +/-0. 15(P <0.01) and 144.31 +/-0.71 (P <0.01) in the control group. The mRNA expression of VEGF and its receptors significantly decreased in gastric cancer cells after cultured with WKN contained serum (P < 0. 01 ).WKN has inhibitory effects on gastric cancer cell growth and mRNA expression of VEGF as well as its receptors KDR and Flt-1.
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Objective To elucidate the effects of thalidomide on the expressions of basic fibroblast growth factor(bFGF) in parental and cisplatin resistant human lung adenocarcinoma cell line A 549 and A 549 DDP Methods RT PCR and lmmunohistochemistry were used to detect the bFGF mRNA and protein expression in A 549 and A 549 DDP Results bFGF mRNA expression levels and protein expressions in A 549 and A 549 DDP were all significant reduced after treatment of thalidomide (6 μg/ml)(all P 0 001).There existed significant differences in A 549 and A 549 DDP among different concentrations of thalidomide on the 5 th day The bFGF protein expression level was consistent with the mRNA expression levels in A 549 and A 549 DDP Conclusion Thalidomide down regulated bFGF expression significantly in A 549 and A 549 DDP cell lines in a dose dependent manner It was suggested that thalidomide not only inhibit tumor angiogenesis but also induce apoptosis
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Objective: To explore the expression of VEGF and P~(53) in acute leukemia(AL) cells,and its value for therapy and prognosis.Method: The VEGF and P~(53) expression was assayed by immunohistochemical staining.Result:The expression rates of VEGF and P~(53) were 69.39% and 55.10%,respectively in 49 AL patients,and were significantly higher than those in the control group.The expression of both VEGF and P~(53) was positive and the complete remission rate was apparently reduced.Conclusion: The high expression of VEGF and P~(53) in AL cells may predict the disease development,chemotherapeutical effect and prognosis.
Clinical Significance
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To investigate the effect of vascular endothelial growth factor (VEGF) on tumor angiogenesis in gallbladder carcinoma.Fifty one patients with gallbladder carcinoma were enrolled as observation group. Thirty healthy people were included as control group. Chemically synthesized siRNA sequences targeting VEGF was transfected with VEGF-siRNA. A blank group (group B), a negative control group (transfected with independent sequence, group C), and an inhibition group (transfected with VEGF siRNA, group D) were established. Physiological saline was set as group A. The expression of VEGF was detected by qRT-PCR. The expression of VEGF protein was detected by Western blot. MVD was used to measure microvessel density. CCK-8, Transwell and flow cytometry were used to detect cell proliferation, invasion and apoptosis.The tumor volume of nude mice and VEGF mRNA expression in group D was significantly smaller than that in group B and C (P<0.05). The MVD density in group B and C was significantly higher than that in group D (P<0.01). The proliferation of cells was detected from the 3rd day, and the proliferation of cells in the blank and negative control groups was faster than that of the inhibition group (P<0.05). The apoptosis rate of the blank group and the negative control group was lower than that of the inhibition group (P<0.001).VEGF is highly expressed in serum of patients with cholangiocarcinoma, it promotes angiogenesis, proliferation and invasion of gallbladder cancer cells, and inhibits apoptosis of tumor cells.
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Objective To study the effect of rh-endostatin on the hypoxia-inducible factor 1α （HIF-1α） and vascular endothelial growth factor （VEGF） expression in mice with lung cancer treated with radiotherapy. Methods Sixty rats were randomly divided into five groups： control group, the large dose single-γ-ray irradiation group, the small dose fraction irradiation group, the large dose irradiation ＋ YH-16 group and the low dose irradiation ＋ YH-16 group. Inhibition rate and the tumor growth curve were observed. Immunohistochemisty was adopted to detect the expression of HIF-1α and VEGF. The correlation between the expression of HIF-1α, VEGF and inhibilion rate was analyzed. Results The expression of HIF-1α or VEGF and inhibition was correlated negatively （r=-7.823,-8.314, respectively ,P〈0.05）. There was no significant correlation between the expression of HIF-1α and VEGF. Conclusion Adminis-tration of YH-16 before radiotherapy can inhibit the HIF-1α and VEGF expression in mice with lung canc-er, and effectively increase the radiosensitivity of lung cancer,.
Key words:
Pulmonary carcinoma; HIF-1α; VEGF; Radiotherapy; Endostatin
Radiosensitivity
Endostatin
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Objective To investigate the effect of thalidomide on vitro liver cancer cell growth,invasion,metastasis and its osteopontin expression. Methods Human hepatoma cell line Hep-G2 was cultured by conventional culture methods. The cell proliferation of Hep-G2 by CCK-8 at different drug concentrations and times was assayed. Cells scratch test was used to verify the impact of thalidomide on the invasion and metastasis ability of Hep-G2. The level of osteopontin expression of Hep-G2 cells was detected by the method of immunocytochemistry and Western blot after the action of different drugs and different reaction time. Result 400 μg / ml of thalidomide could inhibit cell proliferation,compared with the control group in 24,48,72 h( P 0. 01). And the effects positively correlated with reaction time. At the point of 24、48、72 h,compared with the control group,thalidomide significantly inhibited the cell migration( P 0. 01). The immunocytochemistry score was 1. 728 ± 0. 051 in ThD group and 2. 328 ± 0. 245 in control group( P 0. 01). Thalidomide could inhibit the osteopontin expression( P 0. 01) at 24、48、72 h time point,and the effects positively correlated with reaction time. Conclusions Thalidomide can inhibit the proliferation of hepatoma cells and the expression of OPN. And the effects positively correlate with the drug concentration and reaction time.
Osteopontin
Hep G2
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