沉默整合素连接激酶基因对TCA8113舌癌细胞移植瘤生长的影响

发布时间：2018-09-08 08:32

【摘要】：目的研究沉默TCA8113舌癌细胞整合素连接激酶(integrin-linked kinase,ILK)基因的表达,对其下游底物Akt和GSK3β磷酸化状态及对移植瘤生长和自发性转移的影响。方法通过Lipofectamine 2000介导将特异性siILK表达载体和无同源性的对照载体,稳定转染人舌鳞癌TCA8113细胞,分为TCA8113组、TCA8113 vector组和TCA8113 siILK组,将3组细胞分别注入裸鼠皮下构建移植瘤模型,5周后检测瘤质量,对裸鼠肺及瘤组织形态学行常规病理学检查,使用免疫荧光技术检测体外和体内癌细胞p-Akt和p-GSK3β等的表达,并观察瘤组织内血管生成情况。结果 TCA8113组、TCA8113 vector组肺组织均发生自发性转移瘤,TCA8113 siILK组肺组织未发现自发性转移瘤;TCA8113 siILK组移植瘤细胞形态较其他2组体积减小,核分裂象较少,核浆比例缩小;TCA8113 siILK组在体内和体外实验中的p-Akt和p-GSK3β表达较其他2组均明显下降(P0.05);TCA8113 siILK组移植瘤质量较其他2组显著降低(P0.05),抑瘤率分别为84%和92%;TCA8113 siILK组移植瘤血管生成较其他2组也明显减少(P0.05)。结论沉默TCA8113舌癌细胞ILK表达可抑制其下游信号传导分子Akt和GSK3β的磷酸化,并抑制移植瘤血管生成,从而抑制移植瘤的生长。
[Abstract]:Objective to study the expression of integrin-linked kinase (integrin-linked kinase,ILK) gene in TCA8113 tongue cancer cells, and to investigate the effects of silencing the expression of integrin-linked kinase (integrin-linked kinase,ILK) gene on the phosphorylation of Akt and GSK3 尾 downstream substrates and on the growth and spontaneous metastasis of transplanted tumors. Methods Human tongue squamous cell carcinoma (TCA8113) cells were stably transfected with specific siILK expression vector and non-homologous control vector mediated by Lipofectamine 2000. The cells were divided into TCA8113 group (TCA8113 vector group) and TCA8113 siILK group (control group). Three groups of cells were injected into nude mice subcutaneously to construct transplanted tumor model for 5 weeks. The tumor quality was detected by routine pathological examination of lung and tumor histomorphology in nude mice, and the expression of p-Akt and p-GSK3 尾 in cancer cells in vitro and in vivo were detected by immunofluorescence technique. The angiogenesis in tumor tissue was observed. Results spontaneous metastatic tumor was found in lung tissue of TCA8113 group TCA8113 vector group and TCA8113 siILK group. The size of transplanted tumor cells in TCA8113 siILK group was smaller than that in other two groups, and the mitosis was less in TCA8113 siILK group than in the other two groups. The expression of p-Akt and p-GSK3 尾 in TCA8113 siILK group was significantly lower than that in other two groups in vivo and in vitro (P0.05). The tumor quality in TCA8113 siILK group was significantly lower than that in the other two groups (P0.05). The tumor inhibition rates were 84% and 92% in TCA8113 siILK group respectively. Compared with the other two groups, also significantly decreased (P0.05). Conclusion silencing the expression of ILK in TCA8113 tongue cancer cells can inhibit the phosphorylation of Akt and GSK3 尾, and the angiogenesis of xenografts, thus inhibit the growth of transplanted tumors.
【作者单位】：重庆医科大学基础医学院法医生物信息学研究室;重庆医科大学基础医学院法细胞生物学及遗传学教研室分子医学与肿瘤研究中心;重庆医科大学附属口腔医院牙体牙髓科;
【基金】：国家自然科学基金(81172424) 重庆医科大学基础医学院院办课题(0800280016)~~
【分类号】：R739.86

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