| 周世杰,江 姝,赵晓婷.Cyclin Y 和Cyclin X在肺癌细胞株A549中的细胞定位和功能[J].中国肿瘤,2013,22(7):570-574. |
| Cyclin Y 和Cyclin X在肺癌细胞株A549中的细胞定位和功能 |
| The Function Study and Cell Localization of Cyclin Y and Cyclin X in Lung Cancer Cell Line A549 |
| 投稿时间：2013-02-19 |
| DOI：10.11735/j.issn.1004-0242.2013.07.A012 |
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| 中文关键词: Cyclin Y； Cyclin X；肺癌 细胞定位；细胞增殖 |
| 英文关键词:Cyclin Y Cyclin X lung cancer cellular localization cell proliferation |
| 基金项目: |
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| 摘要点击次数: 2022 |
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| 中文摘要: |
| 摘 要：［目的］构建CCNY和 CCNX基因的真核表达载体并观察其在人肺癌细胞株A549中的表达及定位，为进一步探讨Cyclin Y、Cyclin X在肺癌中的细胞定位和功能奠定了基础。［方法］ 以人肺腺癌细胞株H1299 cDNA为模板扩增CCNY和 CCNX基因， 并构建CCNY和 CCNX过表达真核表达载体。应用荧光显微照相及Western Blot方法鉴定该细胞株中Cyclin Y、Cyclin X的定位及表达。［结果］成功构建pEGFP-N1/CCNY和 pEGFP-N1/CCNX真核表达载体。荧光显微照相显示绿色荧光， Western Blot检测证实转染重组质粒细胞表达Cyclin Y、Cyclin X蛋白， Cyclin Y和Cyclin X分别定位于胞膜与胞核。A549-pEGFPN1细胞活性为1.31±0.03，而转染后的A549-CCNY pEGFPN1细胞活性为1.36±0.02， A549-CCNX pEGFPN1细胞活性为1.45±0.05(P<0.01)。［结论］ Cyclin Y和Cyclin X在A549细胞株中定位具有明显的差异，而且胞浆定位的Cyclin X比细胞膜定位的Cyclin Y更能够促进A549细胞增殖。 |
| 英文摘要: |
| Abstract：［Purpose］ To construct pEGFP-N1/CCNY vector and pEGFP-N1/CCNX eukaryotic expression vector，and to explore the location and function of CyclinY/CyclinX in lung caner A549 cell. ［Methods］ CCNY and CCNX genes were amplified from human lung adenocarcinoma cell line H1299 by PCR.The recombinant plasmids pEGFP-N1/CCNY and pEGFP-N1/CCNX were constructed and transfected into A549 cells. The cellular localization and expression of CyclinY and Cyclin X were detected by fluorescence microscopy and Western Blot.［Results］ The recombinant plasmid pEGFP-N1/CCNY and pEGFP-N1/CCNX were constructed successfully. Green fluorescence on the surface of transfected cells was found by fluorescence microscope. Western Blot confirmed Cyclin Y，Cyclin X expression. Cyclin Y and Cyclin X located at cellular membrane and nucleus in recombinant plasmid cell respectively. After transfection，A549-CCNY pEGFPN1 cell viability was 1.36±0.02，A549-CCNX pEGFPN cell viability was 11.45±0.05，which was higher than that in A549-pEGFPN1 (1.31±0.03) (P all<0.01).［Conclusion］ In A549 cell，Cyclin Y and Cyclin X are differently distributed，Cyclin X plays more important role in promoting proliferation than Cyclin Y. |
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