骨肉瘤中Axl抑制凋亡及与凋亡蛋白的相关性

发布时间：2018-04-10 16:31

本文选题：骨肉瘤 + Axl　；参考：《临床与实验病理学杂志》2017年05期

【摘要】：目的探讨受体酪氨酸激酶Axl在骨肉瘤中的抗凋亡作用,分析磷酸化Axl(P-Axl)与抗凋亡蛋白之间的相关性。方法常规培养骨肉瘤细胞株MG63、143B和U2OS,分别设立人重组蛋白Gas6刺激组、Axl siRNA转染组、Gas6刺激+Axl siRNA转染组及各阴性对照组,应用顺铂(DDP)或氨甲喋呤(MTX)致使细胞凋亡后,通过Hoechst 33258染色、MTT实验、Annexin VFIFC等检测法统计分析不同处理组间细胞凋亡率的差异及不同浓度Gas6刺激后骨肉瘤细胞的细胞周期变化;采用免疫组化En Vision两步法检测41例骨肉瘤及18例骨纤维结构不良组织中P-Axl、BCL-2及Bax的表达,分析患者预后与三者的相关性;应用TUNEL染色分析骨肉瘤组织中P-Axl表达水平与细胞凋亡率的相关性。结果 Hoechst 33258、MTT、Annexin V-FIFC等的实验结果一致,均表明骨肉瘤细胞株MG63、143B和U2OS中Gas6通过活化Axl降低DDP或MTX导致的细胞凋亡率(P0.05),Axl siRNA则引起细胞凋亡率增加(P0.05),在Axl siRNA干扰前Gas6预刺激有降低凋亡率的趋势。41例骨肉瘤组织中P-Axl、BCL-2、Bax的阳性率分别为85.4%、70.7%及36.6%;18例骨纤维结构不良组织中三者阳性率分别为11.1%、22.2%及11.1%,两组相比骨肉瘤组织中P-Axl、BCL-2、Bax的表达明显增高(P0.05)。Pearson相关性分析显示BCL-2表达与P-Axl呈显著正相关(r=0.842,P0.000 1);Cox单因素风险回归分析发现BCL-2和Bax表达是影响患者预后的预测因素;TUNEL结果显示P-Axl高表达的骨肉瘤组织中细胞凋亡率明显降低(P0.05)。结论骨肉瘤中Gas6/Axl活化后通过调节凋亡蛋白BCL-2,抑制由DDP或MTX诱发的细胞凋亡,从而促进肿瘤的进展。
[Abstract]:Objective to investigate the antiapoptotic effect of receptor tyrosine kinase (Axl) in osteosarcoma and to analyze the relationship between phosphorylation of P-axl and antiapoptotic protein.The apoptosis rate and cell cycle of osteosarcoma cells stimulated by different concentrations of Gas6 were analyzed by Hoechst 33258 staining and Annexin VFIFC.The expression of P-axln BCL-2 and Bax in 41 cases of osteosarcoma and 18 cases of fibrous dysplasia were detected by immunohistochemical en Vision two-step method.The correlation between P-Axl expression and apoptosis rate in osteosarcoma was analyzed by TUNEL staining.Results the results of Hoechst 33258 Hoechst Annexin V-FIFC were consistent.The results showed that Gas6 in osteosarcoma cell line MG63143B and U2OS decreased the apoptosis rate induced by DDP or MTX by activating Axl. The apoptosis rate of osteosarcoma cell line was increased by P0.05AXL siRNA. There was a tendency of decreasing apoptosis rate by Gas6 prestimulation before Axl siRNA interference. 41 cases of osteosarcoma groupUnivariate risk regression analysis showed that the expression of BCL-2 and Bax was a predictor of prognosis in patients with osteosarcoma. The Tunel results showed that the apoptosis rate of osteosarcoma with high P-Axl expression was significantly lower than that in osteosarcoma with high P-Axl expression (P 0.05).Conclusion the activation of Gas6/Axl in osteosarcoma can inhibit the apoptosis induced by DDP or MTX by regulating the apoptosis protein BCL-2, thus promoting the progression of the tumor.
【作者单位】：中山大学附属第一医院病理科;中山大学附属第一医院骨肿瘤科;
【基金】：国家自然科学基金(81302348)
【分类号】：R738

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