肢体和心脏发育基因在前列腺癌中的表达

Role of limb-bud and heart development expression in prostate cancer

目的:探讨肢体和心脏发育(LBH)基因的表达在前列腺癌中的作用。方法:采用实时荧光定量PCR(q PCR)、免疫组化(IHC)和Western印迹方法检测20例前列腺癌与正常前列腺组织中LBH的表达;构建si RNA-1、si RNA-2和阴性对照序列转染PC-3、LNCa P前列腺癌细胞以沉默LBH的表达,采用CCK-8、Transwell小室法观察细胞增殖、迁移及侵袭能力的变化。结果:q PCR检测结果显示前列腺癌组织中LBHm RNA表达水平明显高于正常前列腺组织(4. 8±2. 4 vs 1. 8±1. 1,P=0. 032); IHC结果显示LBH蛋白在前列腺癌组织中表达阳性率为90. 0%(18/20),正常前列腺组织中为15. 0%(3/20),差异具有统计学意义(P=0. 006); Western印迹结果也显示前列腺癌组织中LBH表达显著高于正常前列腺组织(3. 5±1. 2 vs 1. 3±0. 6,P=0. 019)。干扰LBH表达后,PC-3和LNCa P细胞的增殖能力均显著下降,96 h时,PC-3细胞中si RNA-1和si RNA-2组的A450值显著低于阴性对照组(1. 2±0. 1、1. 1±0. 1 vs 1. 8±0. 2,P <0. 01); LNCa P细胞中si RNA-1和si RNA-2组的A450值也显著低于阴性对照组(1. 4±0. 2、1. 1±0. 2 vs 1. 9±0. 2,P <0. 01)。细胞迁移实验显示,si RNA转染PC-3细胞后,si RNA-1和si RNA-2组透过基底膜的细胞数显著低于阴性对照组[(55. 5±6. 4)、(44. 9±4. 5)个vs (175. 6±25. 8)个,P <0. 05];LNCa P细胞中si RNA-1和si RNA-2组也显著低于阴性对照组[(46. 4±5. 8)、(40. 2±8. 2)个vs (125. 3±13. 5),P <0. 05]。细胞侵袭实验显示,si RNA转染PC-3细胞后,si RNA-1和si RNA-2组透过基底膜的细胞数显著低于阴性对照组[(82. 5±7. 5)、(68. 4±5. 5)个vs (138. 2±10. 7)个,P <0. 05]; LNCa P细胞中si RNA-1和si RNA-2组也显著低于阴性对照组[(46. 4±5. 8)、(23. 1±6. 2)个vs (92. 1±10. 5)个,P <0. 05]。结论:高表达的LBH可能在前列腺癌的发病过程中发挥重要作用,可作为前列腺癌治疗的潜在靶标。

Objective:To investigate the expression and biological function of limb-bud and heart development(LBH) in prostate cancer.Methods:Using real-time quantitative polymerase chain reaction(RT-qPCR),Western blot and immunohistochemistry,we detected the expression of LBH in the prostate cancer and adjacent normal tissues of 20 prostate cancer patients.We transfected the PC-3 and LNCa P cells with si RNA-1,si RNA-2 or negative control si RNA to silence the expression of LBH,and then observed the proliferation,immigration and invasiveness of the cells by CCK-8 and Transwell assays.Results:The expression of LBH m RNA was significantly higher in the prostate cancer than in the normal tissue(4.8 ± 2.4 vs 1.8 ± 1.1,P = 0.032),and so was that of the LBH protein(3.5 ± 1.2 vs 1.3 ± 0.6,P = 0.019) and the positive rate of LBH expression was(90% [18/20]vs 15% [3/20],P = 0.006).Interfering with the LBH expression significantly reduced the proliferation of both the PC-3 and LNCa P cells,with a remarkable decrease at 96 hours in the A450 value of the PC-3 cells transfected with si RNA-1(1.2 ± 0.1) and si RNA-2(1.1 ± 0.1)as compared with that in the negative control group(1.8 ± 0.2)(P < 0.01) as well as in the A450 value of the LNCa P cells(1.4 ± 0.2 and 1.1 ± 0.2 vs 1.9 ± 0.2,P < 0.01).The count of the PC-3 cells that migrated through the polycarbonate membrane was significantly lower in the si RNA-1 and si RNA-2 transfection groups than in the negative control(55.5 ± 6.4 and 44.9 ±4.5 vs 175.6 ± 25.8,P < 0.05),and so was that of the LNCa P cells(46.4 ± 5.8 and 40.2 ± 8.2 vs 125.3 ± 13.5,P <0.05).Cell invasion assay also showed significant lower number of the PC-3 cells penetrating the polycarbonate membrane and matrigel in the si RNA-1 and si RNA-2 transfection groups than in the negative control(82.5 ± 7.5 and 68.4 ± 5.5 vs 138.2 ± 10.7,P <0.05) as well as in that of the LNCa P cells(46.4 ± 5.8 and 23.1 ± 6.2 vs 92.1 ± 10.5,P < 0.05).Conclusion:The overexpression of LBH may play an important role in the development and progression of prostate cancer and serve as a therapeutic target in the treatment of the malignance.