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目的探讨沉默Tiam1基因对直肠癌LYVE-1表达的影响。方法应用免疫组化法检测65例直肠癌组织中Tiaml和Racl蛋白的表达情况并分析其与临床病理特征之间的关系。体外培养人直肠腺癌细胞HR-8348、Colo320、HCT-8,RT-PCR筛选高表达Tiam1的细胞株。构建Tiam1 RNA干扰载体,经酶切鉴定后转染细胞株。用Western blot和免疫细胞化学方法检测干扰Tiam1基因后LYVE-1蛋白表达情况。结果 Tiam1和Rac1蛋白可在直肠癌组织中表达,它们的表达水平均与患者性别、癌肿大小无关(P>0.05),但与组织分化程度、临床分期及淋巴结转移有关(P<0.05)。细胞株HR-8348中Tiam1的表达最强。Tiam1基因干扰后Rac1、LYVE-1蛋白表达水平较干扰前表达有所减弱,尤以LYVE-1蛋白明显,差异有统计学意义(P<0.05)。结论 Tiam1参与直肠癌LYVE-1表达的调节,推测Tiam1通过Rac1影响LYVE-1的表达,诱导直肠癌淋巴管新生,促进淋巴结转移。
Objective To investigate the effect of silencing Tiam1 gene on the expression of LYVE-1 in rectal cancer. Methods The expressions of Tiaml and Racl protein in 65 cases of rectal cancer were detected by immunohistochemistry and their relationship with clinicopathological features was analyzed. Human rectal adenocarcinoma cells HR-8348, Colo320 and HCT-8 were cultured in vitro, and the cell lines highly expressing Tiam1 were screened by RT-PCR. Construction Tiam1 RNA interference vector, identified by restriction enzyme transfected cell lines. The expression of LYVE-1 protein after interfering Tiam1 gene was detected by Western blot and immunocytochemistry. Results The expression of Tiam1 and Rac1 protein in rectal cancer tissues was not correlated with the gender and size of the cancer (P> 0.05), but correlated with the degree of differentiation, clinical stage and lymph node metastasis (P <0.05). The expression of Tiam1 was the strongest in cell line HR-8348. The expression of Rac1 and LYVE-1 protein was weakened after Tiam1 gene interference, especially LYVE-1 protein, the difference was statistically significant (P <0.05). Conclusions Tiam1 is involved in the regulation of LYVE-1 expression in rectal cancer. It is speculated that Tiam1 affects the expression of LYVE-1 through Rac1 and induces lymphangiogenesis in rectal cancer and promotes lymph node metastasis.