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目的观察GABA、GAD65、GABRP在胃癌SGC-7901细胞中的表达及对细胞增殖、细胞周期分布的影响。方法 RT-PCR、IF及Western Blot检测胃癌细胞SGC-7901中GABA、GAD65及GABRP mRNA及蛋白表达;1、10、100μmol/L GABA,0.5、5、50μmol/L Muscimol,0.5、5、50μmol/L Bicucullin分别作用于胃癌细胞SGC-7901 48 h后,新型四唑氮盐(MTS)比色法检测细胞增殖情况,流式细胞仪检测细胞周期分布情况。结果 GAD65、GABRP mRNA及蛋白均表达于SGC-7901细胞中;GABA、GABRP及GAD65蛋白定位于SGC-7901细胞细胞膜和细胞质;与空白组比较,1μmol/LGABA,5、50μmol/L Muscimol作用SGC-7901细胞48 h后,细胞生长的增殖率上升,5、50μmol/L Bicucullin作用SGC-7901细胞48 h后,细胞生长抑制率上升;1μmol/L GABA可导致细胞G0/G1数量的减少,G2/M期数量的增加;3个浓度的Muscimol可导致G0/G1期数量的减少,5、50μmol/L Muscimol可导致细胞G2/M期数量增加;5、50μmol/L Bicucullin可导致G2/M期数量的减少(P<0.05)。结论 GABA及其A受体在SGC-7901细胞中的表达可能促进细胞增殖,扰乱细胞周期进程,促使细胞分裂。
Objective To observe the expression of GABA, GAD65 and GABRP in gastric cancer cell line SGC-7901 and its effect on cell proliferation and cell cycle distribution. Methods The mRNA and protein expressions of GABA, GAD65 and GABRP in SGC-7901 cells were detected by RT-PCR, IF and Western Blot. The expressions of GABA, GABA1, GABA and 0.5, L Bicucullin were respectively treated with gastric cancer cell SGC-7901 for 48 h. The cell proliferation was detected by MTS colorimetric assay. The cell cycle distribution was analyzed by flow cytometry. Results GAD65 and GABRP mRNA and protein were all expressed in SGC-7901 cells. GABA, GABRP and GAD65 proteins were localized in the cell membrane and cytoplasm of SGC-7901 cells. Compared with the blank group, 1μmol / L GABA and 5,50μmol / L Muscimol acted as SGC- The proliferation rate of SGC-7901 cells increased after 48 h treatment with 5 and 50 μmol / L Bicucullin. The growth inhibition rate of SGC-7901 cells increased after 48 h treatment with 5 μmol / L GABA, and decreased with the increase of Gμm / M and M phases. Muscimol at three concentrations could decrease the number of G0 / G1 phases, and Muscimol at 5 and 50μmol / L could increase the number of G2 / M phases. (P <0.05). Conclusion The expression of GABA and its A receptor in SGC-7901 cells may promote cell proliferation, disrupt the cell cycle progression and promote cell division.