基础医学与临床 ›› 2015, Vol. 35 ›› Issue (3): 289-294.

• 研究论文 •    下一篇

RNA干扰Tb舌癌细胞整合素连接激酶基因表达抑制移植瘤生长

幸宇1,邓世雄2,齐进3,姜容1,陈俊霞1   

  1. 1. 重庆医科大学
    2. 重庆医科大学法医学教研室
    3. 重庆医科大学附属口腔医院
  • 收稿日期:2014-07-11 修回日期:2014-11-27 出版日期:2015-03-05 发布日期:2015-03-03
  • 通讯作者: 陈俊霞 E-mail:925760900@qq.com
  • 基金资助:
    RI调控ILK信号通路抑制膀胱癌发生EMT及转移的分子机制研究

RNAi induced integrin-linked kinase silencing in Tb human tongue cancer cells inhibits the growth of xenograft tumor

  • Received:2014-07-11 Revised:2014-11-27 Online:2015-03-05 Published:2015-03-03

摘要: 目的 研究对Tb舌癌细胞的整合素连接激酶(ILK)进行特异性siRNA沉默后,下游信号分子Akt、GSK3β磷酸化状态及对移植瘤生长和自发性转移的变化。方法 建立特异性siILK表达载体和无同源性的对照载体,通过Lipofectamine 2000介导稳定转染人舌鳞癌Tb细胞,然后将Tb、Tb vector和Tb silk 3组细胞分别注入裸鼠皮下构建移植瘤模型,5周后检测瘤大小和质量,对裸鼠肺及瘤组织行常规病理学检查,用免疫荧光技术检测癌细胞的p-Akt和p-GSK3β等的表达,并对瘤组织内血管生成情况进行观察。结果 Tb组、Tb vector组肺组织均发现自发性转移瘤,Tb siILK组肺组织未发现自发性转移瘤;Tb siILK组移植瘤细胞形态较其它二组体积减小,核分裂相较少,核质比例缩小;Tb siILK组在体内和体外实验中的p-Akt和p-GSK3β表达较其它二组均明显下降;Tb siILK组移植瘤质量[(1.68±1.35) g]较Tb组[(3.58±1.04) g]与Tb vector组[(3.64±0.65) g]分别降低了53.0%和53.8%(P<0.05);Tb siILK组移植瘤血管[(5.6±2.2)/视野]生成较Tb组[(15.3±2.3)/视野]和Tb vector组[(14.6±1.4)/视野]也明显减少(P<0.05)。结论 特异性ILK表达沉默抑制Tb舌癌移植瘤的血管生成和瘤细胞增殖,可能与其抑制下游信号传导分子Akt及GSK3β的磷酸化有关,提示ILK有望成为肿瘤治疗的靶基因。

关键词: 整合素连接激酶, 微血管, 移植瘤生长, 人舌鳞癌细胞

Abstract: Objective: To investigate the effects of silencing integrin-linked kinase in Tb human tongue cancer cell on the expression and phosphorylation of its downstream signal molecular Akt and GSK3β and growth and spontaneous metastasis of tumor xenografts. Methods: Target cells were constructed by transfecting specific siILK plasmid and a non-homologous vector negative control into Tb cells. Tb, Tb vector and Tb siILK cells, respectively, were injected into nude mice subcutaneously.Weight and size of the xenograft tumor in nude mice were measured.Morphology of tumor tissue was observed by routine pathology methods.Changes in the expression and phosphorylation of Akt and GSK3β and micro-blood vessels in tumor tissue were detected by immunofluorescence and laser scanning confocal detection, respectively.Results: Phosphorylation of Akt and GSK3β in vivo and vitro was obviously inhibited in Tb siILK group compared with the other groups. Compared with Tb [(3.58±1.04) g]and Tb vector [(3.64±0.65) g]group, the mean tumor mass in Tb siILK group [(1.68±1.35) g]decreased 53.0% and 53.8%,respectively (P<0.05).Microvessel formation was also reduced in Tb siILK group compared to the other groups (P<0.05). Conclusion: Silencing ILK inhibits proliferation and microvessel formation of Tb human tongue cancer tumor xenograft, which may be related with the inhibition of phosphorylation of Akt and GSK3β. The results suggest that ILK could be a therapeutic target protein for tongue cancer.

Key words: integrin-linked kinase, micro vessel, xenograft tumor growth, human tongue squamous cancer cells

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