高迁移率族蛋白A2过表达对大鼠垂体瘤细胞增殖及周期的影响

Effect of high mobility protein A2 overexpression on cell cycle and proliferation of rat pituitary tumor cells

目的 观察高迁移率族蛋白A2（HMGA2）基因过表达对大鼠垂体瘤细胞增殖及周期的影响.方法 制备携带HMGA2基因的质粒（pIRES2-dsRed-HMGA2 cDNA）,转染至大鼠GH3细胞.通过逆转录-聚合酶链反应（RT-PCR）检测细胞内HMGA2 mRNA表达的变化,Western blot检测pIRES2-dsRed-HMGA2 cDNA转染大鼠GH3细胞后HMGA2蛋白表达;细胞计数试剂盒（CCK-8）细胞增殖实验检测HMGA2基因过表达对大鼠GH3细胞增殖的影响;流式细胞仪检测HMGA2基因过表达对大鼠GH3细胞周期的影响.结果 pIRES2-dsRed-HMGA2cDNA转染大鼠GH3细胞24 h后HMGA2 mRNA和蛋白表达水平分别高于对照组的26.40％和36.60％,差异有统计学意义（P＜0.05）,与对照组比较实验组细胞在24、48、72、96 h增殖数量增加（P＜0.05）;流式细胞仪分析显示,实验组的Go/G1期细胞百分率较对照组明显减少[（43.15±0.63）％比（65.33-±0.37）％],而S期较对照组明显增加[（39.57±0.65）％比（18.26±0.54）％,P＜0.05].结论 HMGA2基因过表达促进大鼠垂体瘤细胞增殖,改变细胞正常周期.

Objective To observe the effects of high mobility protein A2 （HMGA2） overexpression on proliferation and cell cycle of rat pituitary tumor cells.Methods The mRNA and protein expression of HMGA2 in rat GH3 cells transduced by pIRES2-dsRed-HMGA2cDNA were examined by reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting.The effects of the HMGA2 overexpression on GH3 cell proliferation and cell cycle were analyzed by cell counting kit-8 （CCK-8） assay and flow cytometry.Results pIRES2-dsRed-HMGA2cDNA transfected rat GH3 cells after 24 h the HMGA2 mRNA and protein expression levels were respectively higher than the control group 26.40％ and 36.60％,the difference was statistically significant （P ＜ 0.05）.To compare with control group the number of pIRES2-dsRed-HMGA2cDNA group cells increased at 24,48,72,96 h （P ＜ 0.05）.Flow cytometry analysis showed that the experimental group,the percentage of cells in G0/G1 phase was significantly reduced compared with the control group,（43.15 ±0.63）％ vs.（65.33 ±0.37）％,while the S phase was significantly increased compared with the control group （39.57 ± 0.65） ％ vs.（18.26 ± 0.54） ％ （P ＜ 0.05）.Conclusion HMGA2 overexpression increases rat GH3 cell proliferation and changes normal cell cycle.