[关键词]
[摘要]
目的: 研究过表达集落刺激因子1 受体(colony stimulating factor-1 receptor,CSF-1R)对人鼻咽癌(nasopharyngeal carcinoma,NPC)6-10B细胞凋亡的抑制作用及其与Bax和Bcl-2 表达之间的关系。方法: 体外利用慢病毒构建的CSF-1R过表达载体LV-CSF1R(16957-1)转染到人鼻咽癌6-10B细胞中,实验分转染组和对照组;采用实时荧光定量PCR 及Western blotting 检测转染后两组细胞中CSF-1R、Bcl-2、Bax的表达情况;CCK-8 法检测两组细胞的增殖;流式细胞术检测两组细胞的凋亡情况。结果:转染组的6-10B细胞中其CSF-1 mRNA表达水平明显高于对照组(7.01±0.23 vs 0.09±0.03,P<0.01); Bax mRNA表达水平显著下调(P<0.01),而Bcl-2 mRNA表达水平显著上调(P<0.01)。转染组的6-10B细胞中CSF-1 蛋白表达水平明显高于对照组;Bax蛋白表达水平显著下调,而Bcl-2 蛋白表达水平稍上调。与对照组相比,转染组6-10B细胞的增殖活力明显提高(P<0.01);凋亡率显著降低[(10.82±0.75) % vs (17.11 ±0.46)%,P<0.05]。结论:过表达CSF-1R 可以通过调节Bax/Bcl-2 之间的比例关系来促进鼻咽癌6-10B细胞的恶性增殖,并抑制细胞凋亡。
[Key word]
[Abstract]
Objective: To investigate the inhibitory effect of Colony stimulating factor- 1 receptor (CSF- 1R) on apoptosis of human nasopharyngeal carcinoma 6-10B cells and its relationship with Bax/Bcl-2 expression. Methods:Lentiviral vector over- expressing CSF-1R LV-CSF1R(16957-1)was constructed and trasfected into 6-10B cells; meanwhile a control group was set; The expression of CSF-1R, Bcl-2 and Bax was detected by Real-time PCR and Western blotting. CCK-8 assay and flow cytometry (FCM) were used to detect the cell proliferation and apoptosis,respectively. Results: The mRNA expression of CSF-1R increased significantly in 6-10B cells of transfection group compared with negative control (NC) group (7.01±0.23 vs 0.09±0.03, P<0.01); The expression of Bax mRNA was significantly down-regulated while the expression of Bcl-2 mRNA was significantly up-regulated in 6-10B cells of transfection group (all P<0.01). The protein expression of CSF-1 in 6-10B cells of transfection group was significantly higher than that of control group, along with the remarkably down- regulated Bax protein expression and slightly up-regulated Bcl-2 protein in transfection group. Compared with NC group, the proliferation vitality of 6-10B cells were extremely increased after transfection (P<0.01); however, the cell apoptosis was significantly reduced ([10.82±0.75]% vs [17.11±0.46]%, P<0.05). Conclusion: Over-expression of CSF-1R could promote the ma-lignant growth and inhibit the apoptosis of nasopharyngeal carcinoma 6-10B cells by regulating the ratio of Bax/Bcl-2 expression.
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[基金项目]
国家自然科学基金资助项目(No.81260348)