| Objective: To study the expression of Ang-2in gastric carcinoma tissue, and itsrelation with pathological stage, To investigate the biological effects of antisense Ang-2cDNA (pEGFP-N1-anti-Ang-2cDNA) in human SGC-7901gastric carcinomacell line.Methods: Immunohistochemical method was used to detect the expression ofAng-2protein in53samples of gastric carcinoma tissues, and western blot technique wasused to the expression of Ang-2protein in10samples of gastric carcinoma and10normal gastric tissues. Using the lipofectamine2000transfection technique, itwere transferred separately two different plasmids induding pEGFP-N1,pEGFP-N1-antisense Ang-2cDNA into an in vitro cultured human gastric carcinoma cell lineSGC-7901. The expression of Ang-2mRNA in53gastric carcinoma tissue specimenswere determined by RT-PCR. Immunohistochemistry was used to detect the expression ofAng-2in those tissues as well.The cell viability was detrmined by MTTassay.Apoptosis was determined by flow cytometry. The three kinds of gastric carcinomacells were subcutaneously injected into30nude mice divided into threegroups freely. After injection, the tumor mass grew and the angiogenesis ofthe tumor could be detected.Results: Ang-2was expressed in different pathological stages of gastriccarcinoma tissue, while normal gastric tissue was unexpressed (F=166.76，P＜0.0001).On RT-PCR and immunohistochemical method,the SGC-7901cells transfected antisense Ang-2gene did not show expression of Ang-2mRNA and its protein.MTT assay showedthe group transfected with antisense Ang-2gene had a much lower proliferative capacitythan other groups (F=10.39，P=0.0024). The group also had an decrease in the rate ofliving cells as measured by flow cytometry Compared with othergroups(χ2=3188.9805，P＜0.0001). The tumor grew slower in transfected mice withantisense Ang-2gene than other groups (the6th day (F=10.18，P=0.0005), the18thday(F=7.8，P=0.0021)), the30th day(F=79.58，P＜0.0001). The average number of newlyformed vessels in the transfected mice with antisense Ang-2gene was smaller than othergroups (the1st week (F=15.18，P＜0.0001), the2nd Week (F=3.50，P=0.0444), the3rdweek（F=39.02，P＜0.0001)).Conclusions: Ang-2plays an important role in the angiogenesis and tumorgrowth, the expression of Ang-2mRNA and its protein in SGC-7901cells can beinhibited by antisense Ang-2gene.The study also show that antisense Ang-2gene cansuppress the in vitro growth of human gastric carcinoma cells. The antisense Ang-2gene may suppress growth and angiogenesis in the human SGC-7901gastric carcinomaapparently. |
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