【摘要】：目的:探討巨噬細胞加帽蛋白(macrophage-capping protein,CapG)對胃癌細胞系遷移和增殖能力的影響。方法:采用real-time PCR方法檢測了4種胃癌細胞AGS、BCG823、PHM82、MNK45中CapG基因的表達情況,選擇低表達并且易轉染的AGS細胞作為研究對象,設計針對CapG的特異性引物并合成重組質粒。構建可以表達CapG的慢病毒包裝系統(tǒng),通過侵染人胃癌細胞系AGS,建立可穩(wěn)定表達CapG的細胞株。通過CCK8實驗分析過表達CapG基因對AGS細胞的生長和增殖能力的影響,細胞劃痕以及Transwell小室實驗分析過表達CapG基因對AGS細胞遷移能力的影響。結果:過表達CapG后,AGS細胞的生長速度略低于對照組,但二者間差異無統(tǒng)計學意義(t=2.424,P=0.073)。劃痕試驗顯示CapG實驗組劃痕間距相對于對照組明顯縮小,兩組平均縮小距離分別為336.99μm和45.54μm,差異有統(tǒng)計學意義(t=14.97,P=0.004)。Transwell試驗顯示CapG實驗組和對照組穿膜細胞數(shù)目分別為176個和70個,CapG實驗組顯著多于對照組,差異有統(tǒng)計學意義(t=40.00,P0.001)。結論:過表達CapG基因對胃癌細胞系AGS細胞的生長和增殖無顯著影響,過表達CapG基因能促進胃癌細胞系AGS細胞的遷移。
[Abstract]:Aim: to investigate the effect of macrophage capsin (macrophage-capping protein,CapG) on migration and proliferation of gastric cancer cell lines. Methods: the expression of CapG gene in AGS,BCG823,PHM82,MNK45 of four gastric cancer cells was detected by real-time PCR method. The low expression and easy transfection AGS cells were selected as the research objects. Specific primers for CapG were designed and recombinant plasmids were synthesized. A lentivirus packaging system expressing CapG was constructed and a stable CapG expression cell line was established by infecting human gastric cancer cell line AGS,. The effects of overexpression of CapG gene on the growth and proliferation of AGS cells were analyzed by CCK8 assay, and the effects of overexpression of CapG gene on the migration ability of AGS cells were analyzed by cell scratch and Transwell chamber experiments. Results: after overexpression of CapG, the growth rate of AGS cells was slightly lower than that of the control group, but there was no significant difference between the two groups (t = 2.424, P < 0.073). Scratch test showed that the distance between scratches in CapG group was significantly smaller than that in control group. The mean reduction distance between the two groups was 336.99 渭 m and 45.54 渭 m, respectively. The difference was statistically significant (t = 14.97, t = 14.97). P0. 004). Transwell test showed that the number of perforating cells in CapG group and control group were 176 and 70, respectively. The number of perforating cells in CapG group was significantly higher than that in control group (t0. 000. 00n P0. 001). Conclusion: overexpression of CapG gene has no significant effect on the growth and proliferation of gastric cancer cell line AGS. Overexpression of CapG gene can promote the migration of gastric cancer cell line AGS.
【作者單位】： 北京大學第三醫(yī)院消化科;北京大學基礎醫(yī)學院病原生物學系;
【基金】：國家自然科學基金(81450024)資助~~
【分類號】：R735.2

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