摘要
目的观察吉非替尼（Gefitinib）和靶向表皮生长因子受体（EGFR）小干扰RNA（siRNA）体内外抑制前列腺癌的效果并探讨其作用机制。方法前列腺癌细胞PC-3转染慢病毒为载体的EGFRsiRNA或Gefitinib（0～10mg／L）处理后，噻唑蓝（MTr）比色法检测细胞生长抑制率，荧光聚合酶链反应（PCR）检测EGFRmRNA表达，Westernblot检测EGFR、丝氨酸蛋白激酶（Akt）、促分裂原活化蛋白激酶（MAPK）和蛋白激酶C（PKC）表达。体内观察EGFRsiRNA和Gefitinib单独或联合抑瘤效果。结果EGFRsiRNA对PC-3细胞转染率达85％，细胞生长抑制率为40％-50％，而Gefitinib细胞生长抑制率呈浓度-时间依赖性。EGFRsiRNA对PC-3细胞EGFRmRNA和蛋白表达抑制率〉90％，明显高于Gefitinib的〈80％（P〈0．01）；EGFRsiRNA显著抑制Akt和MAPK表达，而Gefitinib仅明显抑制Akt表达。Gefitnib单独和联合EGFRsiRNA的抑瘤率分别为53．95％和59．28％，明显高于EGFRsiRNA组的34．83％（P〈0．05）。结论EGFR通路抑制剂能有效抑制前列腺癌生长，其机制主要通过阻断EGFR及其胞内蛋白Akt的表达来实现。
Objective To probe into the inhibitory effects and mechanisms of Gefitinib and small interfering RNAs （ siRNA ） targeting epidermal growth factor receptor （ EGFR ） （ EGFR siRNA ） against hormone independent prostate cancer （HIPC） in vitro and in vivo. Methods After PC-3 cells were transfected with lentivirus mediated EGFR siRNA recombinant or treated with Gefitinib （0-10 mg/L）, MTr was used to measure cell growth inhibitory rate （IR）, fluorescent real-time polymerase chain reaction （PCR） to detect EGFR mRNA level, and Western blotting to assay the expression of EGFR and its intraeellular proteins, such as Akt, MAPK and PKC. Tumor growth was observed when EGFR siRNA or/and Gefitinib were used to treat mice with transplanted tumors. Results The IR of PC-3 cells was 40% -50% by using EGFR siRNA and transfection efficiency was 85%. Gefitnib inhibited the growth of PC-3 cells in a time- and concentration-dependent manner. The expression of EGFR mRNA and protein in PC-3 cells was downregulated over 90% which was obviously higher than 〈 80% in Gefitinib group （ P 〈 0.01 ） ; EGFR siRNA could significantly inhibit the expression of Akt and MAPK, but Gefitinib only significantly inhibit the expression of Akt. In the in vivo study, the tumor growth was inhibited significantly in Gefitinib group （53.95%） or Gefitinib ＋ EGFR siRNA group （59. 28% ） as compared with EGFR siRNA group （34. 83% ,P 〈 0.05 ）. Conclusion EGFR pathways inhibitors could block PC cells growth effectively mainly via the down-regulation of the expression of EGFR and its intracellular protein Akt.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2010年第3期325-327,I0002,共4页
Chinese Journal of Experimental Surgery
关键词
前列腺癌
表皮生长因子受体
RNA小干扰
Prostate carcinoma
Epidermal growth factor receptor
Small interfering RNA
