shRNA-PTTG对A549肺癌细胞株生物学功能的影响

目的探讨shRNA-PTTG对肺癌A549细胞生物学功能的影响。方法采用shRNA技术沉默PTTG蛋白的表达,使用western blot与RT-PCR检测沉默效率,使用western blot检测沉默PTTG后VEGF蛋白的表达影响,使用MTT检测细胞增殖,使用流式细胞仪检测周期。结果 western blot与PCR结果显示shRNA成功沉默PTTG蛋白的表达,且VEGF表达明显下降,MTT结果显示沉默PTTG后,细胞生长明显收到抑制,流式细胞仪结果显示,沉默PTTG后G2-M期明显延缓。结论 PTTG基因具有促进肺癌A549细胞株增殖的作用,且可能是通过VEGF而发挥作用。

Objective To explore the biological functions of shRNA-PTTG on A 549 lung cancer cell. Methods The shRNA technology used to silence the expression of PTTG protein,and then using western blot and rt-pcr detection efficiency;The western blot was also used to detect the expression of VEGF protein after PTTG silenced. MTT was used to detect cell proliferation and using flow cytometry instrument testing cycle. Results Western blot and PCR showed that shRNA success silence the expression of PTTG protein and the expression of VEGF was decreased obviously. MTT and flow cytometry results showed that A 549 cell growth is suppressed obviously and G 2-m period significantly down shifting after PTTG silence. Conclusion PTTG gene can promote the proliferation of A 549 cell lines, and VEGF may be play a role in this procession.