中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
12期
2612-2614
,共3页
范月超%颜丙超%张慧%梅鹏金%陈晨%苗发安%陈洪福%郑骏年%雷霆
範月超%顏丙超%張慧%梅鵬金%陳晨%苗髮安%陳洪福%鄭駿年%雷霆
범월초%안병초%장혜%매붕금%진신%묘발안%진홍복%정준년%뢰정
高迁移率族蛋白A2%过表达%垂体瘤%增殖%细胞周期
高遷移率族蛋白A2%過錶達%垂體瘤%增殖%細胞週期
고천이솔족단백A2%과표체%수체류%증식%세포주기
High mobility protein A2%Over expression%Pituitary tumor%Proliferation%Cell cycle
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.
目的 觀察高遷移率族蛋白A2(HMGA2)基因過錶達對大鼠垂體瘤細胞增殖及週期的影響.方法 製備攜帶HMGA2基因的質粒(pIRES2-dsRed-HMGA2 cDNA),轉染至大鼠GH3細胞.通過逆轉錄-聚閤酶鏈反應(RT-PCR)檢測細胞內HMGA2 mRNA錶達的變化,Western blot檢測pIRES2-dsRed-HMGA2 cDNA轉染大鼠GH3細胞後HMGA2蛋白錶達;細胞計數試劑盒(CCK-8)細胞增殖實驗檢測HMGA2基因過錶達對大鼠GH3細胞增殖的影響;流式細胞儀檢測HMGA2基因過錶達對大鼠GH3細胞週期的影響.結果 pIRES2-dsRed-HMGA2cDNA轉染大鼠GH3細胞24 h後HMGA2 mRNA和蛋白錶達水平分彆高于對照組的26.40％和36.60％,差異有統計學意義(P＜0.05),與對照組比較實驗組細胞在24、48、72、96 h增殖數量增加(P＜0.05);流式細胞儀分析顯示,實驗組的Go/G1期細胞百分率較對照組明顯減少[(43.15±0.63)％比(65.33-±0.37)％],而S期較對照組明顯增加[(39.57±0.65)％比(18.26±0.54)％,P＜0.05].結論 HMGA2基因過錶達促進大鼠垂體瘤細胞增殖,改變細胞正常週期.
목적 관찰고천이솔족단백A2(HMGA2)기인과표체대대서수체류세포증식급주기적영향.방법 제비휴대HMGA2기인적질립(pIRES2-dsRed-HMGA2 cDNA),전염지대서GH3세포.통과역전록-취합매련반응(RT-PCR)검측세포내HMGA2 mRNA표체적변화,Western blot검측pIRES2-dsRed-HMGA2 cDNA전염대서GH3세포후HMGA2단백표체;세포계수시제합(CCK-8)세포증식실험검측HMGA2기인과표체대대서GH3세포증식적영향;류식세포의검측HMGA2기인과표체대대서GH3세포주기적영향.결과 pIRES2-dsRed-HMGA2cDNA전염대서GH3세포24 h후HMGA2 mRNA화단백표체수평분별고우대조조적26.40％화36.60％,차이유통계학의의(P＜0.05),여대조조비교실험조세포재24、48、72、96 h증식수량증가(P＜0.05);류식세포의분석현시,실험조적Go/G1기세포백분솔교대조조명현감소[(43.15±0.63)％비(65.33-±0.37)％],이S기교대조조명현증가[(39.57±0.65)％비(18.26±0.54)％,P＜0.05].결론 HMGA2기인과표체촉진대서수체류세포증식,개변세포정상주기.
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.
