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The development of nanomedicine has recently achieved several breakthroughs in the field of cancer treatment; however, biocompatibility and targeted penetration of these nanomaterials remain as limitations, which lead to serious side effects and significantly narrow the scope of their application. The self-assembly of intermediate filaments with arginine-glycine-aspartate (RGD) peptide (RGD-IFP) was triggered by the hydrophobic cationic molecule 7-amino actinomycin D (7-AAD) to synthesize a bifunctional nanoparticle that could serve as a fluorescent imaging probe to visualize tumor treatment. The designed RGD-IFP peptide possessed the ability to encapsulate 7-AAD molecules through the formation of hydrogen bonds and hydrophobic interactions by a one-step method. This fluorescent nanoprobe with RGD peptide could be targeted for delivery into tumor cells and released in acidic environments such as endosomes/lysosomes, ultimately inducing cytotoxicity by arresting tumor cell cycling with inserted DNA. It is noteworthy that the RGD-IFP/7-AAD nanoprobe tail-vein injection approach demonstrated not only high tumor-targeted imaging potential, but also potent antitumor therapeutic effects in vivo. The proposed strategy may be used in peptide-driven bifunctional nanoparticles for precise imaging and cancer therapy.
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Objective The aim of this study is to explore the changes of expression of integrin αvβ3 in bovine u-terine epithelial cells in vitro induced by estrogen or/and progesterone alone or in combination,and to provide a new refer-ence marker for determining bovine uterine receptivity state. Methods RT-PCR was used to analyze the transcriptional changes of αvβ3 expression in bovine endometrium treated by different concentrations of estrogen,progesterone alone or in combination. Results The expression of integrin αvβ3 reached the highest level when the culture medium was added with progesterone at the concentration of 10 -7mg/mL,and the expression of αv and β3 in the 10 -7mg/mL concentration group was significantly higher than that of the control one(P<0.05). Moreover,the expression of αv was highest in the 10 -10 mg/mL E2group,but the expression of β3 was the lowest in that one. In addition,adding with both estrogen and progester-one,the transcriptional level of integrin αvβ3 was significantly higher than that in the control one. The transcriptional level of αv in the treatment group was significantly higher than that in the control group(P<0.05),but the transcriptional level of β3 in this group was not(P>0.05). Conclusions It can be concluded that integrin αvβ3 can be used as a new poten-tial reference marker gene for detecting the bovine uterine receptive status.
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Objective To explore the role of integrin αvβ3 in the promotion of the development of choroidal neovascularization (CNV) by SDF-1/CXCR4.Methods This study was divided into two parts in vitro and in vivo.As for the in vivo study,a CNV model was induced by laser on C57BL/6J mice,and then assigned into 4 groups:mice with solely CNV modeling as control group,with intravitreal injection of SDF-1 after immediate CNV modeling as SDF-1 group,with intravitreal injection of SDF-1 + CXCR4 inhibitor (AMD3100) after CNV modeling as SDF-1 + AMD3100 group,and mice with intravitreal injection of SDF-1 + αvβ3 inhibitor (SB273005) after modeling as SDF-1 + SB273005 group.CXCR4 and αvβ3 expression levels in laser-induced eyes were quantified by qRT-PCR at time points of day 1,3,5,7,10 and 14 after modeling,and immunofluorescence staining was applied to detect αvβ3 expression in regional CNV and its endothelial cells in the four groups.Finally,OCT was used to observe the height of retinal pigment epithelial (RPE) layers in CNV after treatment in the four groups.Moreover,in the experiment in vitro,Western blot was used to measure the expression of CXCR4 protein of RF/6A cells in normal control group,Si-CXCR4 knockdown group and Si-NC knockdown model group.Meanwhile,the expression of integrin subunit β3 protein was determined in the normal control group,SDF-1 group,SDF-1 + AMD3100group,SDF-1 + Si-NC group and SDF-1 + Si-CXCR4 group.Transwell assay was conducted to detect the migration ability of RF/6A cells in the normal control group,SDF-1group,SDF-1 +AMD3100 group,SDF-1 + SB273005 group.Results On the one hand,the study in vivo,qRT-PCR showed that the expression of CXCR4 and integrin subunit β3 mRNA was up-regulated at first,and then down-regulated with time passed after CNV induction,with the highest expression level of CXCR4 mRNA (4.263 ± 0.464) on day 3,and the peak expression of β3 mRNA (3.678 ±0.364) on day 7 after CNV modeling.The results of immunofluorescence staining showed that the β3 fluorescence intensity of SDF-1 group was significantly enhanced,and the ratio of β3/CD31 was also significantly increased,which both were significantly higher than those of the control group (P ＜ 0.01).However,the β3 fluorescence intensity and β3/CD31 ratio of SDF-1 +AMD3100 group and SDF-1 + SB273005 group were significantly weakened and decreased,respectively (P ＜0.05).OCT showed that the elevation level of RPE layer inSDF-1 group was significantly higher than that in the control group [(135.503 ± 10.301) μm vs.(94.443 ± 12.156) μm](P＜0.05).The height of RPE uplift in SDF-1 + AMD3100 group [(95.283 ±20.062) μm] and SDF-1 + SB273005 group [(99.807 ± 10.403) μm] was significantly decreased (P ＜ 0.05).On the other hand,in experiment in vitro,Western blot showed that the expression levels of integrin β3 in SDF-1 group and SDF-1 + Si-NC group were significantly higher than those in the control group [(1.301 ± 0.043) and (1.273 ± 0.077) vs.(0.244 ± 0.069)] (P ＜ 0.01).The levels of integrin subunit β3 protein in SDF-1 + si-CXCR4 group (0.322 ± 0.042) and SDF-1 + AMD3100 group (0.336 ± 0.077) were significantly down-regulated (P ＜ 0.01).Transwell assay showed that the amount of migrating cells in SDF-1 group increased,which was significantly higher than that of the control group (P ＜ 0.01),while the number of migrating cells in SDF-1 +AMD3100 group and SDF-1 + SB273005 group was significantly decreased.Conclusion Integrin αvβ3 can promote the development of CNV by mediating SDF-1/CXCR4 signaling in endothelial cells.
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Objective To evaluate the effect and the anti-tumor mechanism of nucleoside combination on human hepatoma cell line Bel-7402 with Arg-Gly-Asp sequence labeled by ultrasmall superparamagnetic iron oxide (RGD-USPIO).Methods The tumor cells Bel-7402 of logarithmic phlyhase were divided into experimental group and control group,treated with 1 mmol/L nucleoside combination and 1640 medium respectively.The two group were co-cultured for 48 h,and were added RGD-USPIO and co-cultured for 6 h.Then the two groups were proceeded with MR scanning,and the signal intensity of T2WI were measured.After extraction of the total RNA and protein of experiment group and control group,the expression of integrin avβ3 was detected using real-time PCR and Western blot.Results The T2WI signal intensity of experimental group (997.35±42.83) was higher than that of control group (241.05±15.36,t 28.79,P＜0.01).Compared with control group,the expression of integrin αvβ3 mRNA in experimental group was (0.22±0.02) times (t=4.50,P＜0.01).According to Western blot,the protein bands of experimental group were relatively lighter than that of control group,the expression of integrin αvβ3 in experimental group was lower (t =11.88,P＜0.01).Conclusion Nucleoside combination has anti-tumor effect by inhibiting integrin ligand-receptor binding,and the anti tumor mechanism may be related to the induction of tumor cell apoptosis.MR molecular probes can conveniently and accurately evaluate the anti-tumor effect of nucleoside combination on Bel-7402 cells.
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Objective To synthesize a molecular probe targeted to human hepatoma HepG2 cells with high expression of integrin αvβ3 (RGD-PEG-VSOP) and evaluate its MRI efficacy in vitro.Methods RGD-PEG-VSOP was characterized and analyzed by 1H NMR and TEM.MTT test was used to evaluate its biological safety.In vitro experiments at the cellular level,the targeting effect of RGD-PEG-VSOP to integrin was assessed,meanwhile the nontargeted nanoparticles were used as controls.Results TEM showed that the nanoparticles were spherical and uniform in size,with a relatively high r1 relaxivity of 1.37 mM-1S-1.MRI showed the signal intensity of the HepG2 cells treated with RGD-PEG--VSOP was significantly higher than that of the HepG2 cells treated with PEG-VSOP (P＜0.05).Conclusion RGD-PEG-VSOP has positive T1 contrast effect.At the cellular level,the RGD-PEG-VSOP nanoparticles have the characteristics targeted to integrin αvβ3.
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Objective: To study the effect of lentivirus-mediated integrin αVβ3-shRNA on tumor growth of mice with lung cancer xenograft. Methods: Lung cancer tissue, paracancer tissue and normal tissue were collected and integrin αVβ3 expression was detected; BALB/c nude mice were selected, divided into integrin αVβ3 knockdown group (KD group) and negative control group (NC group), and inoculated with cells stably infected by integrin αVβ3-shRNA lentivirus and cells stably infected by negative control-shRNA lentivirus, respectively, the growth of tumor tissue was continuously observed, and the number of apoptosis cells as well as the expression of angiogenesis, apoptosis and invasion genes in tumor tissue were detected. Results: mRNA content and protein content of integrin αVβ3 in lung cancer tissue were significantly higher than those in paracancer tissue and normal tissue; increasing trend of tumor tissue volume of KD group was weaker than that of NC group, and tumor volume at various points in time of KD group was lower than that of NC group; mRNA contents and protein contents of VEGF, FGF, EGF, Bcl-2, MMP-9, MMP-12 and MMP-13 in tumor tissue of KD group were lower than those of NC group, and apoptosis index as well as mRNA content and protein content of Bax were higher than those of NC group. Conclusions: The expression of integrin αVβ3 increases in lung cancer tissue, and lentivirus-mediated integrin αVβ3-shRNA can inhibit tumor growth of mice with lung cancer xenografts.
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OBJECTIVE@#To study the effect of lentivirus-mediated integrin αVβ3-shRNA on tumor growth of mice with lung cancer xenograft.@*METHODS@#Lung cancer tissue, paracancer tissue and normal tissue were collected and integrin αVβ3 expression was detected; BALB/c nude mice were selected, divided into integrin αVβ3 knockdown group (KD group) and negative control group (NC group), and inoculated with cells stably infected by integrin αVβ3-shRNA lentivirus and cells stably infected by negative control-shRNA lentivirus, respectively, the growth of tumor tissue was continuously observed, and the number of apoptosis cells as well as the expression of angiogenesis, apoptosis and invasion genes in tumor tissue were detected.@*RESULTS@#mRNA content and protein content of integrin αVβ3 in lung cancer tissue were significantly higher than those in paracancer tissue and normal tissue; increasing trend of tumor tissue volume of KD group was weaker than that of NC group, and tumor volume at various points in time of KD group was lower than that of NC group; mRNA contents and protein contents of VEGF, FGF, EGF, Bcl-2, MMP-9, MMP-12 and MMP-13 in tumor tissue of KD group were lower than those of NC group, and apoptosis index as well as mRNA content and protein content of Bax were higher than those of NC group.@*CONCLUSIONS@#The expression of integrin αVβ3 increases in lung cancer tissue, and lentivirus-mediated integrin αVβ3-shRNA can inhibit tumor growth of mice with lung cancer xenografts.
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Objective To investigate the synthesis, in vivo biodistribution of 99Tcm-HYNIC-PEG4-E[PEG4-c (RGDfk)] 2 (99 Tcm-Galacto-RGD2), and its potential usage for targeted imaging of mice orthotopic glioma.Methods 99Tcm-Galacto-RGD2 was synthesized straightforward and its radiochemical purity and stability and distribution in mice were analyzed.MicroSPECT-CT imaging was done in a mice orthotopic glioma model, which had been set up with U87MG cells, after administration of 99Tcm-Galacto-RGD2.Region of interest (ROI) of glioma was drawn on SPECT-CT section images to quantify tumor uptake (％ ID/cm3).Glioma was harvested for pathological examination.Linear-regression was used to analyze the relationship between integrin αvβ3 and tumor uptake (％ID/cm3).Results The radiochemical purity of 99Tcm-Galacto-RGD2 was (97.7 ±0.8)％ and stable in vitro.Hynic-Galacto-RGD2 could specifically bind to integrin αv β3 of tumor cells with a IC50 of (18 ± 3) nmol/L.After tail vein injection, 99Tcm-Galacto-RGD2 was rapidly discharged from the blood, liver, kidneys and had a relative low concentration in normal brain tissue.MicroSPECT-CT imaging demonstrated that, after 60 min of injection, this drug was well uptaken by glioma tumor than that after 30 min (t =7.13 ,P ＜0.05), and the tumor to normal brain tissue (T/B) uptake ratio of 99Tcm-Galacto-RGD2 was 13.92± 3.43.Injection of HYNICGalacto-RGD2 2 min prior to 99Tcm-Galacto-RGD2 injection extensively reduced the uptake of radioactive drug in tumor tissue (t =11.36, P ＜ 0.05).Bland-Altman analysis showed that tumor volume based on SPECT-CT imaging measurement had almost same value with the tumor reference volume (95％ CI =-11.94％-11.92％).In addition, the tumor uptake of 99Tcm-Galacto-RGD2 and cellular integrin αvβ3 expression level had a linear relationship (R2 =0.896).Conclusions Stable 99Tcm-Galacto-RGD2 can be synthesized easily and is applicable for microSPECT-CT imaging analysis of orthotopic glioma in mice together with the evaluation of integrin αvβ3 level in tumor.
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Objective The purpose of this study was to prepare RGD4C modified ferritin nanocages (RGD4C-FRT) for targeted drug delivery to rat hepatic stellate cells (HSC-T6). Methods RGD4C modified human H-chain ferritin was expressed and purified. Doxorubicin (Dox) was encapsulated into the cavity of RGD4C-FRT through ion channels, which resulted in RGD4C-FRT-Dox. The target of RGD4C-FRT-Dox to HSC-T6 was detected using fluorescence microscopy. Results Transmission electron microscopy showed that RGD4C-FRT was hollow spherical-structured with uniform size and good dispersion. The average particle diameters of RGD4C-FRT and RGD4C-FRT-Dox were 12.57 nm and 20.13 nm , respectively. The drug encapsulation efficiency and loading percentage of RGD4C-FRT-Dox were 77.32% and 15.88% respectively. RGD4C-FRT-Dox was significantly uptaken by HSC-T6, and the uptake could be blocked by the empty carrier RGD4C-FRT. Conclusion RGD4C-FRT-Dox can specifically target HSC-T6. Further animal experiments are needed to inspect its therapeutic effect on liver fibrosis.
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Objective To observe the MVD and integrin αvβ3 mRNA expression changes in tumor -bearing rats by using endostar combined with radiotherapy ,and to explore the potential synergy mechanism .Meth-ods We randomly divided the tumor -bearing rats into four groups:control group(NC),endostar group(ES), radiation treatment group ( RT) and endostar combined radiotherapy group ( ES +RT) .Tumor inhibition rate was calculated every other day .Microvessel density and αvβ3mRNA were texted by immunohistochemical or real time PCR in each group.Results (1)ES+RT group showed the most obvious inhibition rate;(2)Compared with NC group,microvessel density of RT was increased ,but decreased in ES group and ES +RT group significantly ( P<0.05);(3)Compared with NC group,αvβ3 mRNA expression of RT group was increased,while decreased in ES group and ES+RT group,and ES +RT group displayed a greater decrease when compared to ES group ( P<0.05).Conclusion Endostar combined radiotherapy can inhibit the growth of cancer and the expression ofαvβ3mRNA,improve the disorders of tumor vascular network .It may be one of the mechanisms of increasing radi-osensitization.
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This study examined the impact of luteinized unruptured follicle (LUF) on endometrial receptivity.The menstrual cycle of 17 LUF patients (LUF group) and 13 ovulatory women (control group) was monitored by measuring LH level in urine and by ultrasonic examination.An endometrial biopsy at the sixth to tenth day after LH surge was taken in all the patients.The expressions of endo-metrial ER,PR and integrin avβ3 were immunohistochemically determined.At the same time,the serum levels of E2 and P were detected by chemiluminescence.The results exhibited that (1) The mean serum P level in LUF group (7.32±2.56 ng/mL) was significantly lower than that in control group (11.17±3.17 ng/mL) (P＜0.01).But there was no significant difference in the mean serum E2 levels between LUF group (179.35±81.60 pg/mL) and the control group (198.58±75.23 pg/mL) (P＞0.05); (2) The mean expression intensities of ER,PR in endometrium of LUF group (183.86±2.43,167.94±3.04) were significantly higher than those in control group (109.35±6.31,105.98±4.07)(P＜0.01); (3) The mean expression intensities of integrin αvβ3 in endomtrium of LUF patients (114.90±11.38) were significantly lower than those in control group (191.34±1.82) (P＜0.01); (4)The change profile of integrin αvβ3 expression in the endometrium of LUF patients was in positive relation with serum P level (r=0.77,P＜0.01),but bore no significant relationship with serum E2 level (r=0.01,P＞0.05).It was concluded that the depression of serum P levels in LUF patients was closely related to the failure of the down-regulation of ER and PR,and the low expressions of integrin αvβ3 also suggested that the delayed implantation and the impaired endometrial receptivity had impact on embryonic implantation.
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To investigate the expression of osteopontin (OPN) and its receptor integrin αvβ3 in the placental tissue from pregnant women complicated with preeelampsia,the expression of OPN and αvβ3 in the placenta of the pregnant women with preeclampsia and healthy pregnant women was detected by immunohistochemistry,Western blotting and RT-PCR. Our results showed that OPN and αvβ3 protein were expressed in the placenta from normal pregnant woman and those with preeclampsia. OPN was located in the placental syncytiotrophoblasts and the cytoplasm of capillary endothelial cells and integrin αvβ3 was mainly expressed on the surface of trophoblast cells. Expression of OPN and integrin αvβ3 in the placental tissue from preeclampsia subjects was significantly lower than that from the control group (P＜0.05). Compared with the control group,expression of OPN in the placental tissue from preeclampsia group was significantly lower (P＜0.05) but there was no significant difference in the expression of αv and β3 between the preeclampsia group and the controls. It is concluded that OPN and its receptor integrin αvβ3 may be involved in the pathogenesis of preeclampsia.