Tiam1过表达调控喉癌生长及侵袭转移的体内外实验研究

Investigation of Tiam1 overexpression on the growth,invasiveness and metastasis of laryngeal carcinoma in vitro and in vivo

目的前期研究显示T淋巴瘤侵袭转移诱导因子1(Tiam1)与喉癌临床预后密切相关。为了进一步明确Tiam1对喉癌生物学行为的影响,通过上调Tiam1在喉癌细胞中的表达,观察Tiam1上调后对喉癌细胞体内外增殖和侵袭转移能力的影响。方法通过质粒转染构建Tiam1稳定过表达的喉癌Hep-2细胞(Hep-2/Tiam1);本实验以pc DNA3(Mock)质粒作为转染对照。实验共分3组:空白对照组(Hep-2),转染pc DNA3(Mock)的Hep-2细胞株(Hep-2/Mock)和稳定转染Tiam1基因的Hep-2细胞株(Hep-2/Tiam1)。通过MTT、划痕实验及Transwell侵袭实验观察Tiam1过表达后对Hep-2细胞体外增殖、迁移及侵袭能力的影响。通过构建喉癌裸鼠移植瘤动物模型,进一步在体内水平观察Tiam1对喉癌的增殖及侵袭转移能力的影响。结果MTT实验显示Hep-2/Tiam1与对照组Hep-2及转染空载体的Hep-2比较,各组之间细胞的增殖能力无明显差异(P>0.05)。划痕实验显示Hep-2/Tiam1细胞的迁移距离明显多于另两组(P<0.05)。Transwell侵袭实验发现穿过聚碳酸酯膜的Hep-2/Tiam1较其他两组明显增多(P<0.05)。Hep-2细胞的裸鼠移植瘤动物模型的成瘤率达100%。实验组Hep-2/Tiam1移植瘤的体积及重量与其他两组相比均无明显差异(P>0.05),但颈淋巴结转移率显著增加(P<0.05)。结论Tiam1过表达后对喉癌Hep-2细胞体内外的增殖能力无明显影响,但能增强Hep-2细胞体内外的侵袭转移能力,提示其在喉癌的进展中发挥了重要作用。

Objective Our previous study has demonstrated that T lymphoma invasion and metastasis inducing factor 1(Tiam1)is correlated with the prognosis of laryngeal carcinoma.Hence,we will upregulate the Tiam1 expression in cell lines of laryngeal carcinoma,and observe the effect of Tiam1 upregulation on the proliferation,migration and invasion of laryngeal carcinoma in vitro and in vivo.Methods The Hep-2 cell line of stable high Tiam1 expression(Hep-2/Tiam1)was successfully obtained via plasmid transfection.Plasmid pc DNA3 selected for Mock transfection,blank control group(Hep-2),Hep-2/Mock and Hep-2/Tiam1 were included.The related experiments(MTT assay,scratch assay and Transwell invasion test)were carried out to observe the effects of Tiam1 upregulation on the proliferation,migration and invasion of laryngeal carcinoma cell line Hep-2 in vitro.Then,the metastatic xenograft mouse model of laryngeal carcinoma was constructed.The related experiments were also carried out to observe the effects of Tiam1 upregulation on the proliferation and invasion of Hep-2 in vivo.Results Compared with the group of Hep-2 and cell line with null vector transfection,MTT assay revealed that there were no significant statistical differences in proliferation among 3 groups(P>0.05).However,Tiam1 upregulation led to significantly increased migration(P<0.05)and invasion(P<0.05)of laryngeal carcinoma Hep-2 cells.The tumor formation rate of laryngeal carcinoma metastatic xenograft mouse model was 100%.There were no significant statistical differences among 3 groups on the tumor volume and tumor weight of xenografted tumors(both P>0.05).The cervical lymph nodes metastasis rate in the group of Hep-2/Tiam1 was significantly increased(P<0.05).Conclusion Upregulation of Tiam1 has no effect on the proliferation of Hep-2 cells in vitro and in vivo,but can increase the migration and invasion of Hep-2 cells in vitro and in vivo,suggesting that Tiam1 contributes to the laryngeal carcinoma progression.