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@#Objective To observe the growth of xenografted tumor in nude mice after DDX46 expression decreased, and to further study the role of DDX46 in the development and progression of esophageal squamous cell carcinoma. Methods DDX46-shRNA mediated RNAi was applied to silencing DDX46 in Eca-109 cells. Twenty-five female BALB/c nude mice were divided into 3 groups: an experiment group (DDX46-shRNA-LV, n=10), a control group (Control-LV, n=10) and a blank control group (Het-1A, n=5). The prepared Eca-109 cells of DDX46-shRNA-LV and Control-LV were subcutaneously injected into the right armpit of mice (4×106 cells per mouse), while Het-1A cells were subcutaneously injected into the bilateral armpits of mice (4×106 cells per side). Tumor growth was monitored twice a week on the 14th day after injection. Tumor volume was measured with calipers, in vivo imager to observe the fluorescence of each group. Further, western blotting analysis was used to detect the changes of apoptosis signaling molecules in xenografted tumor after DDX46 silence. Results The growth of xenografted tumor in nude mice was significantly slower in the DDX46-shRNA-LV group than that in the Control-LV group throughout the study period (P<0.001). Western blotting analysis showed that silencing DDX46 effectively suppressed the expression of DDX46, and upregulated the expression of cleaved Caspase-3 and cleaved PARP-1 in xenografted tumor (P<0.01). Conclusion DDX46 is involved in the development and progression of esophageal squamous cell carcinoma, and the silence of DDX46 expression can inhibit the growth of esophageal squamous cell carcinoma, which probably by positive regulation of apoptosis signaling pathway.
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Background and purpose:This study was to investigate the effect of miRNA-486 on the growth of human colorectal cancer cell line SW620 xenograft in nude mice and to explore the possible mechanism of action. Methods:Eighteen mice were randomly divided into three groups, including the experimental group, the negative control group and the blank control group. Each group contained 6 mice. The SW620 cell line was inoculated subcutaneously into nude mice to establish the model of human colorectal cancer xenografts. Peritumoral injection of miRNA-486 overexpres-sion plasmid, or blank vector and PBS were performed every 3 days. The volumes of subcutaneous tumors in each group of inoculated mice were compared. Then mice were sacrificed 3 weeks after infection. Immunohistochemistry and Western blot were used to measure the expression of neuropilin-2 (NRP2).Results:The growth rate of tumors in the experimental group was significantly lower than that in the negative control group and the blank control group. After 21 days, the size of transplanted tumors in the experimental group nude mice was (0.32±0.12) cm3, that in the negative control group was (0.77±0.31) cm3, and that in blank control group was (0.82±0.18) cm3. Tumor mass in the experimental group was sig-nificantly smaller than that in the other two groups (P=0.006<0.05). Tumor mass in the experimental group was (0.40±0.08) g, significantly smaller than that in the negative control group (0.75±0.18) g and in the blank control group (0.79±0.18) g (P=0.008<0.05). Compared with the expression of NRP2 in other groups, the growth of tumor in the experimental group de-clined (P=0.000<0.05).Conclusion:Colorectal cancer cell line SW620 xenografted tumor in nude mice can be suppressed after injection of miR-486, which may decrease the expression of NRP2.
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Objective To observe the effect of rhTRAIL on survivin gene expression of human lung adeno-carcinoma A549 xenografted tumor in nude mice,and investigate the possible inhibitory mechanism of rhTRAIL on the implanted-tumor growth.Methods The solid tumor model was formed in nude mice with human lung adeno-carcinoma cell line.A549.24 mice were randomly divided into the four groups,rhTRAIL single treated group (1 μg/mL),rhTRAIL combined with cisplatin (DDP) treated group,cisplatin treated (1.5mg/kg) and 0.9％ sodium chloride injection(NS) control group.The rhTRAIL and DDP were injected once every other day by intraperitoneal injection to mice in the treated groups,lasting eight times,the same volume of saline solution was injected to the control group.After these,mice were killed and dissected completely.The expression level of survivin mRNA and protein in the tumor tissues was detected by real-time polymerase chain reaction (RT-PCR) and immunohistochemistry,respectively.And the expression of survivin gene in serum of each group was tested by ELISA.Results The expression levels of survivin mRNA in implanted-tumor tissues in rhTRAIL,rhTRAIL combined with DDP,DDP and NS group were (48.7 ± 2.5) ％,(53.1 ± 4.6) ％,(99.1 ± 5.3) ％ and (95.6 ± 3.7) ％,respectively.While the protein expressions of survivin gene in those groups were (0.319 ± 0.025),(0.483 ± 0.058),(0.635 ± 0.041) and (0.619 ± 0.017),respectively.Moreover,the serum levels of survivin were (71.9 ± 7.05),(80.26 ± 10.80),(112.75 ± 15.41) and (105.03 ± 20.37),respectively.The data showed that the expression levels of rhTRAIL and rhTRAIL combined with DDP group were lower than that of DDP-treated group or the NS control group (P ＜ 0.0 5).Compared with the rhTRAIL combined with DDP group,the survivin gene expression level of rhTRAIL-single treated group decreased a little lower,but the difference was not significant (P ＞ 0.05).Conversely,the survivin gene level was increased to some degree compared with the NS control group,and uniformly there was no significant difference (P ＞ 0.05).Conclusion rhTRAIL can downregulate the expression level of survivin gene of human lung adeno-carcinoma A549 xenografted tumor in nude mice.It may be one of the possible inhibitory mechanisms of rhTRAIL on the implanted-tumor growth that rhTRAIL can downregulate survivin gene expression and promote tumor cell apoptosis.
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Objective: To investigate the effects of curcumin combined with oxaliplatin on the human colon cancer cells LoVo xenografted tumor in nude mice and to explore the mechanism. Methods: Nude mice were implanted with human colon cancer LoVo cells. All tumor-bearing mice were randomly divided into four groups and treated with vehicle, 50 mg/kg curcumin, 25 mg/kg oxaliplatin, and their combination (50 mg/kg curcumin + 25 mg/kg oxaliplatin) by ip injection once every other day individually. After continuous administration of drug treatment for 11 times, the weights of nude mice were recorded, the stripping tumor weight was monitored, and the tumor volume and tumor inhibitory rates were calculated. The enucleation of eyeball for taking blood and blood routine examination were carried out and the function of liver and kidney was detected. Tumor cell cycle and apoptosis rate were assayed by flow cytometry. The pathological morphology of tumor was analyzed by HE staining. The apoptosis related gene expression was detected by RT-PCR. Results: Tumor inhibitory rates of curcumin, oxaliplatin, and curcumin + oxaliplatin groups were 59.47%, 55.49%, and 70.56%, respectively. Curcumin combination with oxaliplatin did not influence the blood system, liver, and kidneys in nude mice. Combination of curcumin and oxaliplatin could effectively inhibit the tumor growth (P < 0.05), interfere with cell cycle arresting at S and G2/M phases (P < 0.05, 0.01), and promote the expression of bax (P < 0.01) in tumor-bearing nude mice. Conclusion: Combination of curcumin and oxaliplatin could synergistically inhibit the growth of LoVo colonic xenografts in nude mice.
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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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Background and purpose:There were reports that radiotherapy might intensify angiogenesis and result in the resistance of tumor to radiotherapy.Endostar,a new type rh-endostatin,may specifically inhibit angiogenesis and tumor growth,so it may reduce the resistance to radiotherapy.This study was to evaluate the inhibitory effect of Endostar in combination with radiation compared with cisplatin(DDP) plus radiation on lung adenocarcinoma A549 in BALB/c nude mice.Methods:We established A549 lung adenocarcinoma xenograft models.When the largest diameter of tumor reached 1.0 cm,all mice were randomized into 4 groups(n=6):control,radiation alone,radiation plus DDP,radiation plus Endostar.We measured the largest diameter and the vertical diameter of the tumors at different time points after treatment.At the 16th day,mice were executed and the tumors were subjected to removal for the analysis of apoptosis,and the expression level of basic fibroblast growth factor(bFGF) mRNA by reverse transcription-polymerase chain reaction(RT-PCR) and vascular endothelial growth factor(VEGF) were also detected by immunohistochemistry.Results:The tumor growth velocity of control,radiation alone,radiation plus DDP,radiation plus Endostar were(162?6)%,(131?8)%,(104?7)% and(108?11)% respectively(P0.05).The apoptosis rates of control,radiation alone,radiation plus DDP,radiation plus Endostar were(12.2?1.1)%,(16.5?0.8)%,(24.4?1.5)% and(20.5?1.9)%,respectively(P