ABSTRACT
Objective To investigate the effects of anti-sense hTERT on the expression of VEGF and the receptor in SGC7901 cells, and study the influence in angiogenesis and progression in gastric carcinoma. Methods SGC7901 cell line was transfected by the recombinant virus containing sense and anti-sense hTERT cDNA. Then the expression of VEGF-C and Flt-4 was observed with RT-PCR, distribution of cell cycles was determined with flow cytometry. The expression of VEGF-C and Flt-4 protein was assessed with immunohistochemistry. Result The distribution of cell cycle of antisense hTERT transfected SGC7901 cells was changed, and the mRNA and protein expression of VEGF-C and Flt-4 was significantly down-regulated. Conclusion Anti-sense hTERT can act as an agent for inhibiting VEGF-C and Flt-4 mRNA and protein level, and it blocks tumor angiogenesis and lymphogenous metastasis.
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Objective To study the effect of transfection of antisense MBD1 gene eukaryotic expression vector on the expression of MBD1 gene in human cholangiocarcinoma cell line QBC-939.Methods The(constructed) antisense MBD1 gene eukaryotic expression vector was transfected into the human(cholangiocarcinoma) cell line QBC-939 using lipofectamine transfection reagents,and positive cell clones were obtained using G418 selection after transfection.The constructed recombinant vector was transfected into(QBC-939) cells successfully and was confirmed by amplifying the exogenous neo~R gene with PCR method.The expression level of MBD1 gene mRNA and protein was detected by RT-PCR and FCM methods respectively.Results Following the transfection,the MBD1 gene mRNA level in human cholangiocarcinoma cell line QBC-939 decreased from 0.912?0.022 to 0.215?0.017,and the MBD1 gene protein level also(decreased) from(80.19?5.05)% to(35.11?4.05)%.There were very significant differences on the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression vector transfection group(P
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Objective To probe into the mechanisms of bone morphogenetic protein-2 (BMP-2) in osteosarcoma and to provide basis for gene therapy. Methods A 1.0 kb cDNA fragment of human BMP-2 gene was inserted reversely into PDOR and Human antisense BMP-2 retrovirus expression vector was constructed. The recombinant retroviral vector was transfected into human osteosarcoma cells OS-9901 with liposome AMINE that expressed abundant BMP. Positive cell clones were selected with G 418. The expression of BMP and proliferating cell nuclear antigen (PCNA) was determined by immunohistochemical ABC methods. The osteosarcoma cell cycle was analysed by flowcytometry, and the changes were observed by electronmicroscope. Results The expression of cellular BMP and PCNA in the transfected osteosarcoma cells decreased obviously (t=24.01， 26.09, respectively, P
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Objective To develop a new way to prevent vascular anastomotic intimal hyperplasia, we applied surgical suture soaked in c-myc antisense oligodeoxynucleotide solution to perform vascular anastomosis in rabbits. Methods8/0 sutures (coated Vicryl) soaked in antisense, sense and mismatched c-myc oligomer solution respectively were applied to accomplish vascular anastomoses in rabbits with interposition of external jugular veins between common carotid arteries. Twenty-four animals were randomly divided into four groups: control group, antisense group, sense group and mismatched group. Four weeks later, angiography was made to scrutinize the patency of anastomosis and specimens were harvested for microscopy, the image was digitalized by computer video analysis system, the intimal thickness and area, medial thickness and area and their ratios were calculated and analyzed statistically among groups. Resultsall the anastomoses were normal and patent. The intimal thickness, intimal/medial ratio and intimal area, intimal/medial area ratio in antisense group were significantly lower than those in other groups(P
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The aim of this study was to construct antisense of ZNRD1 encoding gene, and to transfect it into SGC7901/VCR cells, to seek measures to overcome multidrug resistance of gastric cancer cells. ZNRD1 cDNA amplified by PCR was confirmed by DNA sequencing, and then inserted into the multiple cloning site of the expressing vector pcDNA3 1 + with molecular cloning technique. The recombinant vector was identified by endonuclease digestion. Antisense recombinant vector was transfected into SGC7901/VCR cells using lipofectamine. Flow cytometric analysis was used to detect adriamycin (ADM) accumulation in SGC7901/VCR cells. The results showed that a fragment was obtained by PCR, and its sequence was consistent with ZNRD1 cDNA reported in the literature. After antisense recombinant vector having been transfected into SGC7901/VCR cell, ADM concentration in such cells was increased. The above results indicated that the antisense vector ZNRD1 could enhance the intracellular accumulation of ADM in SGC7901/VCR cells, which might be of potential treatment value.