整合素连接激酶对裸鼠前列腺癌的影响

Effect of integrin-linked kinase on the growth of prostate cancer in nude mice

目的:探讨应用整合素连接激酶(ILK)特异siRNA敲除ILK基因对人DU145细胞系裸鼠原位前列腺癌生长及进展的影响。方法:将合成的ILK特异siRNA转染人DU145细胞系,应用逆转录PCR检测ILK在转录水平的表达情况;使用Westernblot方法检测ILK在蛋白水平的表达情况。初步探讨敲除ILK基因后对人DU145细胞的黏附、侵袭性的影响及其对细胞骨架结构的影响。分别将ILKsiRNA和对照DU145细胞注入裸鼠皮下,每5天1次,连续4周,检测裸鼠前列腺癌肿瘤的大小、分化程度、凋亡及增殖状态来观察ILK对前列腺癌生长的影响。结果:siRNA显著降低了ILK基因的表达,ILKmRNA及蛋白的表达分别被抑制87%与81%。与对照组相比,实验组DU145黏附力及侵袭力显著下降,实验组细胞骨架结构破坏明显。裸鼠种植DU145细胞后5周,实验组与对照组相比,细胞在裸鼠种植部位形成的肿瘤体积明显减小[(18.1±1.3)mm3vs(157.6±9.7)mm3,P<0.01)],分化程度变好[Gleason评分(5.8±0.2)vs(8.8±0.3),P<0.05],凋亡指数增加[(64.75±5.87)vs(38.57±3.45),P<0.01],增殖指数减小[(42.75±4.76)vs(88.57±7.57),P<0.01]。结论:动物实验表明,特异性的抑制ILK能抑制人DU145细胞的黏附及其侵袭能力,破坏DU145细胞骨架的形成,诱导前列腺肿瘤细胞的凋亡及抑制肿瘤细胞的增殖,从而在一定程度上抑制了肿瘤的生长及进展。

Objective：To investigate the effect of the integrin-linked kinase（ILK） small interfering RNA（siRNA） on prostate cancer in nude mice by orthotopic injection of human cell line DU145.Methods：The cultured human cell line DU145 was knocked down for ILK using a siRNA.Cellular ILK expression was quantified by RT-PCR and Western blot analysis.Moreover,cell attachment,invasiveness and microfilament dynamics assays were performed.Furthermore,the impact of the ILK siRNA on the prostate cancer was tested using a nude mice model in which prostate cancer was induced by orthotopic injection of human prostate cancer cell line DU145.Gross tumor volume of prostate in nude mice,cell differentiation,the state of apoptosis and proliferation were tested after 5 weeks of injection.Results：The expression of ILK was suppressed significantly by siRNA,cellular mRNA and protein of ILK decreased 87% and 81% separately.The knockdown of ILK also induced the attachment and invasiveness of DU145 cell growing down.The tumor volume,cell differentiation,apoptosis index and proliferation index of prostate in nude mice of ILK siRNA orthotopic injection model were significantly smaller,better,increased and decreased separately than those in control group.Conclusion：Targeting inhibition of ILK not only decreases attachment and invasiveness of human DU145 cells,but also suppresses the growth and development of prostate cancer of orthotopic injection human DU145 cell line model in nude mice.