抑制信号转导与转录激活因子3对人前列腺癌细胞DU145生长的作用研究*

网络出版时间：2015-10-13网络出版地址：http://www.cnki.net/kcms/detail/52.5012.R.20151013.1147.008.html

张启芳， 禹文峰， 官志忠

(贵州医科大学 分子生物学重点实验室， 贵州 贵阳550004)

[摘要]目的： 研究抑制信号转导与转录激活因子3(STAT3)表达对人前列腺癌细胞DU145生长的影响。方法： 用慢病毒pFLU-EGFP STAT3 shRNA(shRNA)质粒转染人前列腺癌细胞DU145构建抑制STAT3表达的稳定细胞株作为shRNA组，转染空载体作为空白对照(mock)组，采用荧光定量聚合酶链式反应(qPCR)和免疫印迹(Western blot)方法检测STAT3的mRNA和蛋白的表达水平，用结晶紫染色法检测抑制STAT3表达对细胞生长的影响；用Matrigel细胞侵袭实验检测抑制STAT3表达对DU145细胞迁移侵袭能力的影响。结果： 与mock组相比，shRNA组STAT3 mRNA和蛋白表达水平显著下降，约为mock组的15%和20%，shRNA组DU145细胞的生长数目明显减少，仅为空白对照组的41.8%；shRNA组DU145细胞对基底膜的侵袭能力降低，抑制率为mock组52.9%。结论： 抑制STAT3表达可降低前列腺癌细胞生长和迁移能力。

[关键词]信号转导与转录激活因子3； 前列腺癌细胞； 细胞生长； 细胞侵袭

[基金项目]*国家自然科学

[中图分类号]R737.25； R34-33[文献标识码] A

The Effect of AbrogatingSTAT3 on Human Prostate Cancer Cell Growth

ZHANG Qifang, YU Wenfeng, GUAN Zhizhong

(KeyLaboratoryofMolecularBiology,GuizhouMedicalUniversity,Guiyang550004,Guizhou,China)

Abstract[] Objective: To investigate the effect of STAT3 silencing on growth and invasion of human prostate cancer cells. Methods: Lentiviruses expressing STAT3-specific shRNA were used to infect human prostate cancer DU145 cells to generate a stable cell line as shRNA group, non-specific shRNA as mock group. STAT3 expression was assessed by qPCR at transcriptional level and Western blot at protein level. The effect of STAT3 inhibition on cell growth was determined by crystal violet staining. Matrigel invasion assay was used to evaluate the effect of STAT3 inhibition on cell invasion.Results: Compared to mock group, STAT3 expression were silenced about 15% at transcriptional level and 20% at protein level. Cell growth was significantly delayed in shRNA group, about 41.8% of mock control. STAT3 silencing significantly impaired cell invasion ability and invaded cells were about 52.9% of mock group. Conclusions: Inhibiting STAT3 expression hinders growth and invasion of human prostate cancer cells.

[Key words] signal transducer and activator of transcription; prostate cancer cells; cell growth; cell invasion

男性癌症死亡病例中，前列腺癌位居第2，雄激素剥夺疗法最初能有效治疗部分前列腺癌患者，但大多数患者2~3年后复发，最终发展为势抵抗性前列腺癌(castration-resistant prostate cancer，CRPC)。对于CRPC患者，尚无有效的治疗方法[1]。信号转导与转录激活因子3(signal transducer and activator of transcription 3,STAT3)是白介素-6(interleukin-6,IL-6)家族细胞因子、生长因子、G偶联蛋白受体和Toll-样受体等信号通路的交汇点[2]。许多人类恶性肿瘤中均出现持续性激活的STAT3蛋白[3-5]，诱导细胞增殖和抑制抗肿瘤免疫反应等，导致肿瘤发生[2]；……