1.Advance and prospects of nanoscale diagnosis and therapy technology for tumors
Chinese Journal of Cancer Biotherapy 2006;0(05):-
Nanomaterials,endowed with unique properties,bring new chance for early diagnosis and therapy of tumors and show a bright future.In recent years,research in in vitro detection techniques of nanoparticle-based early tumor biomarkers,in vivo dynamic multimodel molecular imaging techniques,nanoscale effects of nanopartilce-based nano-imaging and treatment integrated technology,nanoscale slow-release drugs,and nanoscale drug delivery devices has made great progress.However,the long term interaction between nanomaterials and human body still remains poorly understood,the application of nanoscale therapy technology in clinical medicine still faces new challenges.In this paper,we review the main advances of tumor nanoscale diagnosis and therapy,with the aim to improve the development of tumor nanoscale diagnosis and therapy technology in China.
2.Influence of the inhibitory oligonucleotides on the immune status of BXSB lupus mice
Zhiming LI ; Peng HUANG ; Jun CHEN ; Daxiang CUI
Chinese Journal of Primary Medicine and Pharmacy 2010;17(19):2598-2600
Objective To research whether the inhibitory oligonucleotides could improve the immune status of the BXSB mice. The purpose was to provide a valuable direction for treating systemic lupus erythematosus. Methods 3-month-old BXSB lupus mice were divided into three groups (including inhibitory oligonucleotides group, saline group and blank control group). The 24 hours urine proteins were determined before treatment. After treatment,the urine protein, anti-dsDNA, peripheral blood lymphocytes apoptosis and immune complex in renal glomeruli were measured. Results Before treatment,the urine proteins had no statistical differences among the three groups ( P > 0.05 ).After treatment,the urine protein, anti-dsDNA levels in inhibitory oligonucleotides group were significantly lower than that of control group(P < 0.01 ). Apoptosis of peripheral blood lymphocytes in inhibitory oligonucleotides group was significantly higher than that of control group( P < 0.05 ) ,immune complex in renal glomeruli in inhibitory oligonucleotides group was significantly lower than that of the other groups ( P < 0.05 ). Compared with saline group, inhibitory oligonucleotides had prolonged life period of BXSB mice ( P < 0.01 ). Conclusion The inhibitory oligonucleotides could improve immune status of BXSB, and could put off the disease progression.
3.Fluorescent supermagnetic nanoparticles-labeled adipose-derived mesenchymal stem cells in the three-dimensional culture system
Pinlei LV ; Yuehan SU ; Daxiang CUI ; Zheng WANG
Chinese Journal of Tissue Engineering Research 2015;(1):44-48
BACKGROUND:Fluorescent magnetic nanoparticles with the properties of quantum dots and magnetic particles have good biocompatibility, and can label cels effectively through endocytosis. OBJECTIVE:To validate the feasibility of fluorescent magnetic nanoparticles in labeling human adipose-derived mesenchymal stem cels. METHODS:Healthy human adipose tissue was extracted and adipose-derived mesenchymal stem cels were isolatedin vitro by type I colagenase digestion. Passage 6 cels were incubated with the fluorescent magnetic nanoparticles overnight. Prussian blue staining and laser scanning confocal microscope were used to observe labeled adipose-derived mesenchymal stem celsin vitro after co-culturing with fluorescent magnetic nanoparticles. The tracing effect of labeled adipose-derived mesenchymal stem cels in vivo was detected by fluorescence imaging system. RESULTS AND CONCLUSION:Prussian blue staining showed that the fluorescent magnetic nanoparticles dispersed in the cytoplasm of adipose-derived mesenchymal stem cels in the form of blue particles. Under the laser scanning confocal microscope, the nuclei of adipose-derived mesenchymal stem cels were dyed blue by Hoechest33258, and the cytoplasm was dyed green. The fluorescence imaging results showed that labeled human adipose-derived mesenchymal stem cels had good imaging results. Therefore, as an efficient tracer, the fluorescent magnetic nanoparticles can label human adipose-derived stem cels in vitro and provide a new method for transplantation and transformation of adipose-derived mesenchymal stem cels.
4.Apoptosis in Human Osteosarcoma Cell Line SOSP-9607 Induced by Caspase-6
Yong DING ; Qingyu FAN ; Daxiang CUI ; Dianzhong ZHANG ; Jianning YIN
Chinese Journal of Cancer Biotherapy 1996;0(04):-
Objective:To explore caspase-6's effect on the apoptosis of osteosarcoma cell line SOSP 607. Methods: The expression level of caspase-6 in the osteosarcoma cell line SOSP-9067 was examined by RT-PCR method. The adeno-virus adv5 vector with caspase-6 gene was constructed and transferred into the osteosarcoma cell line SOSP-9607 by lipo-fection. The cell survival rate after transfection was assayed by MTT method. The cell morphological changes were observed by microscope and electron microscope, the apoptosis of transferred cells were examined by gel electrophoresis. Results:No expression of caspase-6 was examined in the osteosarcoma cell line SOSP-9607. A high expression of caspase-6 was identified by RT-PCR after the transfection. The cell growth curve declined after transferring caspase-6. Electrophoresis of DNA displayed the apoptosis ladder. Conclusion: Transferring caspase-6 into the osteosarcoma line SOSP-9607 may inhibit the growth of the osteosarcoma cell line SOSP-9607 and this effect might be achieved by inducing apoptosis.
5.Cloning, expression and purification of novel gene NBEAL1 and its relationship with pathological grades of glioma
Chenchen BAO ; Hao YANG ; Na LI ; Bin LIU ; Hua SONG ; Ping SHENG ; Guohan HU ; Daxiang CUI
Chinese Journal of Cancer Biotherapy 2010;17(1):77-81
Objective: To construct the expression plasmid of a novel gene human NBEAL1 (neurobeachin like 1), and to study its relationship with the pathological grades of glioma. Methods: Total RNA of human glioma cell line U251 was extracted. NBEAL1 expression plasmid pGEX-KG/NBEAL1 was constructed and transferred into E. coli BL21. Recombinant NBEAL1 protein was induced by IPTG and further purified by GST affinity chromatographic column. The purity of recombinant NBEAL1 protein was examined by Western blotting analysis. A NBEAL1 protein specific monoclonal antibody was prepared and was used to study the relationship of NBEAL1 expression with pathological grades of glioma. Results: The NBEAL1 gene fragment was successfully cloned into pGEX-KG expression plasmid and verified by DNA sequencing. The recombinant NBEAL1 protein was expressed in inclusion bodies, with a yield of more than 30% of total bacterial proteins; the purity of purified NBEAL1 protein was above 95%. Western blotting analysis confirmed that the purified protein containing GST tag and NBEAL protein. NBEAL1 protein was lowly expressed in normal brain tissues and highly expressed in low grade glioma tissues; and the expression of NBEAL1 decreased with the increase of glioma malignancy. Conclusion: The NBEAL1 protein has been successfully cloned, expressed and purified. NBEAL1 protein expression in glioma tissues is negatively associated with the pathological grades of glioma.
6.Antitumour effects of survivin antisense oligonucleotide carried by nano-high-polymer on colorectal cancer
Zhai CAI ; Zhou LI ; Zonghai HUANG ; Suzhen FANG ; Daxiang CUI ; Feng GAO
Chinese Journal of Tissue Engineering Research 2009;13(47):9393-9397
BACKGROUND: In recent years, nano-carriers have been regarded as the most promising technologies for breakthrough the bottleneck of gene transfer. Polyamidoamine dendrimer (PAMAM) is a kind of new nanometer material. PAMAM can transfer target gene to the cell with high efficiency and lower toxic both in vivo and in vitro.OBJECTIVE: To evaluate the antitumour effects of survivin antisense oligonucleotide (Survivin-asODN) carried by PAMAM on colorectal cancer transplanted subcutaneously in nude mice.DESIGN, TIME AND SETTING: An in vivo experiment regarding tumor gene therapy was performed from February to August in 2008 at the Laboratory of Bionanometer Engineering, Research Institute of Micro/nanometer Science & Technology of Shanghai Jiao Tong University and Central Laboratory of Zhujiang Hospital of Southern Medical University.MATERIALS: Human colorectal cancer cells SW620 were from Shanghai Cell Institute of Chinese Academy of Sciences.PAMAM dendrimer was offered by the Bionanometer Engineering Laboratory, Research Institute of Micro/nanometer Science & Technology, Shanghai Jiao Tong University. Lipofectamine ~(TM)2000 was purchased from Invitrogen, USA. Survivin-asODN was synthesized by Shanghai Bioengineering Company.METHODS: The PAMAM and cation liposome were respectively mixed with Survivin-asODN to generate the transfection complex carrying antisense gene. The shape of the complex was observed by transmission electron microscope, the particle size was determined by laser particle size analysator and the zeta potential was measured by an analytical tool. The encapsulating efficiency and release progress in vitro were determined by ultraviolet spectrophotometer in centrifuging method. Human colorectal cancer cells SW620 at logarithmic phase were inoculated into the abdominal region of 18 Blab/C nude mice subcutaneously to produce transplanted tumor models in colorectal cancer nude mice, which were randomly divided into 3 groups: liposome, PAMAM and blank control groups. They were injected respectively with Hposome-survivin-asODN complex,PAMAM-survivin-as ODN transfection complex and seroculture liquid. The volumes of tumor were surveyed in the 2 groups.Western blotting method was used to determine the survivin gene expression in the transplanted tumor tissue.MAIN OUTCOME MEASURES: Particle size, zeta potential, gene loading level, encapsulation efficiency, release rate of cationic liposome-survivin-asODN complex and PAMAM-survivin-asODN complex, as well as survivin expression rate and apoptosis rate after transfection, inhibition rate of the transplanted tumor growth, Survivin protein expression and activity in the transplanted tumor cells.RESULTS: The particle size of PAMAM-survivin-asODN complex was smaller (P < 0.01), but the zeta potential was greater (P < 0.05), compared with liposome-survivin-asODN. There was no significant difference between PAMAM and liposome groups in terms of gene loading rate and transfection efficiency. DNA release lasted for 14 days for PAMAM, but only 5 days for liposome.After colorectal cancer cell transfection, survivin protein expression was lower, but apoptosis rate was higher, in the PAMAM-survivin-asODN complex than in the liposome-survivin-asODN complex (P < 0.05).CONCLUSION: PAMAM facilitates delivery of Survivin-asODN into transplanted colorectal cancer cells SW620. As a result,survivin protein expression was decreased, and apoptosis rate was increased in vivo which inhibited transplanied tumour growth.
7.Overexpressed Gene YA61 Cloned from Human Gastric CarcinomaCell SGC-7901 and Its Sequencing
Jinrong ZHAO ; Xiaojun YAN ; Fengchan HAN ; Daxiang CUI ; Yu HOU ; Quanjian YAN ; Chengzhi SU
Progress in Biochemistry and Biophysics 2001;28(1):99-102
To clone overexpressed gene from human gastric carcinoma cell SGC-7901, DDRT-PCR technique is used with human gastric epithelial cell GES-1 as control. After cloned into pGEM -T vector, YA61, one of the overexpressed genes, was analyzed by dot blot and was sequenced then. The sequence gotten was then compared to GenBank data and analyzed by NCBI ORF Finder. Dot blot results showed that the gene YA61 was overexpressed in human gastric carcinoma cell SGC-7901. NCBI's sequence similarity search indicated that the gene YA61 was a new gene sequence. Open reading frame analysis demonstrated that the gene YA61 had one complete open reading frame. In conclusion, the gene YA61 was a new gene sequence that was overexpressed in human gastric carcinoma cell SGC-7901.
8.Inhibitory effects of carbon nanotube-PAMAM-anti-survivin oligonucleotide compounds on proliferation of HepG2 cells
Bifeng PAN ; Daxiang CUI ; Ping XU ; Feng GAO ; Rong HE ; Xiaogang YOU ; Jun SHAO ; Hao YANG
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective:To investigate the efficiency of carbon nanotube(CNT)-PAMAM mediated entrance of anti-survivin oligonucleotide into HepG2 cells,and its effects on the proliferation of HepG2 cells.Methods:CNT-PAMAM-anti-survivin oligonucleotide compounds were prepared and characterized by AFM and 1% agarose gel electrophoresis analysis.TEM was used to observe the distribution of CNT-PAMAM-ASODN compounds in HepG2 cells.CNT-PAMAM-ASODN compounds were added into the medium and co-cultured with HepG2 cells for 24 h,48 h,72 h,and 96 h at 37℃,5% CO_2.MTT method was used to detect the effects of ASODN and CNT-PAMAM-ASODN on the proliferation of HepG2 cells.Results:CNT-PAMAM-ASODN compounds were successfully synthesized via AFM and agarose gel electrophoresis.TEM showed that the compounds were located in the cytoplasm.When CNT-PAMAM-ASODN(1.0 ?mol/L)and ASODN(1.0 ?mol/L)were used for a 48 h culture,the inhibitory rates of HepG2 cells were(45.97?4.28)% for CNT-PAMAM-ASODN compounds group,(9.33?0.85)% for ASODN group,and(6.37?0.69)% for CNT-PAMAM group.CNT-PAMAM-ASODN compounds at 1.5 ?mol/L inhibited HepG2 cells by(70.22?7.25)%,and the inhibitory effects were in a time-and concentration-dependent manner.There was statistical difference between experiment group and control group(P
9.Inhibitory effects of BRCAA1 gene silencing on gastric cancer MGC-803 cells and its possible mechanism
Bin LIU ; Daxiang CUI ; Tong DU ; Zhiming LI ; Hua SONG ; Hao YANG ; Chenchen BAO ; Hui GAN
Chinese Journal of Cancer Biotherapy 2006;0(06):-
Objective:To investigate the inhibitory effect of breast cancer-associated antigen 1(BRCAA1)gene silencing on gastric cancer MGC-803 cells and the related mechanism.Methods:Plasmid shRNA-BRCAA1 and shRNA-N were constructed and transfected with FuGene HD into gastric cancer cell line MGC-803.The transfection efficiency was examined using fluorescent microscope 24 h later.The total RNAs was extracted 48 h 'after transfection and the expression of BRCAA1 and GAPDH gene were analyzed by real-time PCR.The cell proliferation was assessed by MTT assay 24 h,48 h,and 72 h after transfection.The cell apoptosis was determined by Annexin V-PE/TAAD.The expression of Rb,Bax, Bcl-2 and BRCAA1 proteins was analyzed by Western blotting 48 h after transfection.Results:We found that the transfection efficiency of shRNA-BRCAA1 was(81.2?2.6)%24 h after transfection.Forty-eight hours after transfection with shRNA-BRCAA1 the expression of BRCAAI mRNA decreased by 61.4%;the inhibition rate of MGC-803 cells growth was 45.0%.The cell apoptosis rate of shRNA-BRCAA1 transfection group was significantly higher than those of untransfected group and mock plasmid transfected group([14.4?1.6]%vs[5.4?2.0]%,[4.4?2.5]%,P
10.In vitro Anti-viral Activity of the Total Alkaloids from Tripterygium hypoglaucum against Herpes Simplex Virus Type 1
Zhe REN ; Chuanhai ZHANG ; Lianjun WANG ; Yunxia CUI ; Renbin QI ; Chongren YANG ; Yingjun ZHANG ; Xiaoyi WEI ; Daxiang LU ; Yifei WANG
Virologica Sinica 2010;25(2):107-114
Herpes simplex virus type 1 (HSV-1) is a commonly occurring human pathogen worldwide. There is an urgent need to discover and develop new alternative agents for the management of HSV-1 infection. Tripterygium hypoglaucum (level) Hutch (Celastraceae) is a traditional Chinese medicine plant with many pharmacological activities such as anti-inflammation, anti-tumor and antifertility. The usual medicinal part is the roots which contain about a 1% yield of alkaloids. A crude total alkaloids extract was prepared from the roots of T. hypoglaucum amd its antiviral activity against HSV-1 in Vero cells was evaluated by cytopathic effect (CPE) assay, plaque reduction assay and by RT-PCR analysis. The alkaloids extract presented low cytotoxicity (CC_(50) = 46.6 μg/mL) and potent CPE inhibition activity, the 50% inhibitory concentration (IC_(50)) was 6.5 μg/mL, noticeably lower than that of Acyclovir (15.4 μg /mL). Plaque formation was significantly reduced by the alkaloids extract at concentrations of 6.25 μg/mL to 12.5 μg/mL, the plaque reduction ratio reached 55% to 75% which was 35% higher than that of Acyclovir at the same concentration. RT-PCR analysis showed that, the transcription of two important delayed early genes UL30 and UL39, and a late gene US6 of HSV-1 genome all were suppressed by the alkaloids extract, the expression inhibiting efficacy compared to the control was 74.6% (UL30), 70.9% (UL39) and 62.6% (US6) respectively at the working concentration of 12.5μg/mL. The above results suggest a potent anti-HSV-1 activity of the alkaloids extract in vitro.