Pokemon基因干扰对泌乳素腺瘤细胞的影响及其分子机制

目的:探讨原癌基因pokemon干扰对泌乳素腺瘤细胞的影响及其分子机制,为可能的泌乳素腺瘤分子靶向治疗提供依据。方法:从泌乳素腺瘤患者手术切除的肿瘤组织中分离培养原代肿瘤细胞,利用siRNA技术构建pokemon稳定干扰泌乳素腺瘤细胞系(siRNA1、siRNA2和siRNA3),采用荧光定量PCR及Western blot验证pokemon的表达;同时运用CCK8检测pokemon基因干扰对泌乳素腺瘤细胞增殖的影响。采用Western blot检测在泌乳素腺瘤组织及siRNA稳定干扰细胞中pokemon对下游cyclin-A、p14ARF和p21蛋白表达的影响。结果:3株pokemon基因干扰泌乳素腺瘤细胞均构建成功。与空白质粒对照组比较,干扰组的细胞增殖能力均较对照组明显减弱(P均<0.05),且siRNA2干扰效率最高。与正常泌乳素腺组织比较,泌乳素腺瘤组织中pokemon蛋白表达明显增加(P=0.042),而其下游蛋白p14ARF、cyclin-A以及p21表达均显著降低(P均<0.05)。与空白质粒对照组比较,在siRNA2干扰组细胞中p14ARF、cyclin-A和p21蛋白表达水平均明显升高(P均<0.05)。结论:pokemon基因干扰可上调泌乳素腺瘤细胞中肿瘤相关抑制因子的表达,提示pokemon可能作为泌乳素腺瘤治疗的潜在靶位点。

Pokemon gene induced prolactinoma progression and related molecular mechanisms

OBJECTIVE:Prolactinoma is a common intracranial tumor wihch always leads to serious clinical outcomes. This study aims to confirm the association between the proto-oncogene pokemon and prolactinoma progression, and further explore the molecular mechanisms. METHODS:Tumor tissues were collected from patients with prolactinoma. The primary prolactinoma cells were digested from the tissues and cultured with proper conditions. The expression of pokemon gene was knocked-down by siRNA transfection, and the proliferation of prolactinoma primary cells was analyzed by CCK8 assay. The expression level of pokemon related genes, cyclin-A, p14ARF and p21 in tissues and prolactinoma primary cells were examined by Western blot analysis. RESULTS:Expression of pokemon in prolactinoma primary cell was successfully knocked-down by siRNA transfection. The results showed that siRNA2 showed the highest efficiency of inhibiting the proliferation ability of prolactinoma primary cells. Cyclin-A, p14ARF and p21 were down-regulated by pokemon in both prolactinoma tissues and primary cells. CONCLUSION:Pokemon could improve prolactinoma progression by reducing the expression level of tumor inhibition factors, cyclin-A, p14ARF and p21.