T淋巴瘤侵袭转移诱导因子1过表达对人结直肠癌细胞生物学特性的影响

Effects of Tiam1 overexpression on biological characters of human colorectal cancer cells

目的:应用已构建稳定过表达T淋巴瘤侵袭转移诱导因子1(T lymphoma invasion and metastasis,Tiam1)的人结直肠癌细胞株,探讨Tiam1对人结直肠癌细胞生物学特性的影响。方法:考马斯亮蓝染色以及扫描电镜观察Tiam1基因转染前后细胞形态学改变;细胞周期、MTT法、平板克隆形成实验、Transwell小室检测细胞体外增殖、迁移及侵袭能力;裸鼠皮下成瘤实验检测细胞体内增殖能力。结果:相对于HT29/mock细胞,HT29/Tiam1细胞多呈梭形,伪足增多、纤长;HT29/Tiam1细胞处于S期的细胞比例更高(t=19.546,P=0.000),细胞增殖能力增强(F=177.125,P=0.000),克隆形成率增高(t=3.222,P=0.032),HT29/Tiam1细胞穿过微孔膜(t=4.832,P=0.001)和Matrigel(t=3.779,P=0.005)的细胞数均增加,差异具有显著性;HT29/Tiam1和HT29/mock两组细胞的裸鼠体内生长差异从第15天开始出现,到第30天时,HT29/Tiam1细胞组的肿瘤体积是HT29/mock细胞组的2.3倍,差异具有统计学意义(F=53.040,P=0.002)。结论:Tiam1稳定过表达能够促进结直肠癌细胞的增殖、运动及侵袭能力,提示Tiam1在结直肠癌的发生、发展中发挥着重要作用,有望为结直肠癌的临床治疗提供新的靶点。

Objective To explore the effect of Tiaml overexpression on the biological behaviors of human colorectal cancer cells （CRC）. Methods The human CRC lines under the established stable overexpression of Tiaml were studied. Cell morphology was detected before and after transfection by commassie blue staining and scanning electron microscope. The proliferation in vitro of CRC was tested by cell cycle, MTr and plate colony formation assay, the migration and invasion ability of CRC was tested by Transwell assay. The proliferation ability in vivo was studied by induced subcutaneous tumors of nude mice. Results Compared with HT29/mock cells, HT29/ Tiaml cells formed as spindle, the pseudopodia increased and elongated. The proportion in S phase of HT29/Tiaml was higher （t = 19.546, P = 0.000）, the proliferation ability enhanced （F = 177.125, P = 0.000）, colonies formation ratio increased （t = 3.222, P = 0.032）. The number of HT29/Tiaml cells acrossing the microporous membrane （t = 4.832, P = 0.001）and Matrigel（t = 3.779, P = 0.005）all raised. On the fifteenth day, the growth deference between the HT29/Tiamlcells and HT29/mock cells in nude mice in vivo occurred. Till the thirtieth day, the size of the tumors in HT29/Tiaml cell group were 2.3 times as large as that in HT29/mock cell group （F = 53.040, P = 0.002）. Conclusions Tiaml stable overexpression can promote the proliferation, migration and invasion ability of CRC which indicates its important role in carcinogenesis and evolution of colorectal cancer. Tiaml may represent a new therapeutic target for colorectal cancer.