Shp2对舌癌细胞恶性生物学行为的调控作用

The regulative effect of Shp2 on malignant biological behavior of tongue carcinomas cell

目的探讨Src同源磷酸酪氨酸磷酸酶2(Shp2)在舌癌组织中的表达水平及其对Cal-27细胞增殖、转移、侵袭、凋亡等恶性生物学行为的影响。方法采用免疫组织化学法检测舌癌组织中Shp2的表达情况;采用RNA干扰技术靶向沉默Cal-27细胞中Shp2的表达;采用噻唑蓝(MTT)法检测Cal-27细胞24、48、72、96 h的增殖活性;采用流式细胞术检测Shp2对Cal-27细胞凋亡的影响;采用蛋白质印迹(Western blot)法检测Shp2对Cal-27细胞中Bax、p53和Bcl-2蛋白表达的影响;采用Transwell小室侵袭实验和划痕实验检测Shp2对Cal-27细胞体外侵袭和迁移能力的影响。结果舌癌组织中Shp2的阳性表达率明显高于癌旁正常组织(P﹤0.01);Shp2-si RNA Cal-27细胞的增殖、侵袭和迁移能力均低于Cal-27细胞(P﹤0.05)。与空白对照组比较,Shp2-si RNA组和阴性对照组的Cal-27细胞凋亡率均有所升高,差异均有统计学意义(P﹤0.05)。Shp2-si RNA组中的p53、Bax蛋白表达水平均高于空白对照组和阴性对照组,Bcl-2蛋白表达水平低于空白对照组和阴性对照组,差异均有统计学意义(P﹤0.05)。结论 Shp2在舌癌细胞中具有促癌基因的活性,可促进Cal-27细胞的增殖、迁移、侵袭,抑制其凋亡。

Objective To investigate the expressions of Src homology phosphotyrosine phosphatase 2 （Shp2） in tongue carcinomas tissues and elucidated the effect of Shp2 on Cal-27 cell biological behavior. Method The expressions of Shp2 in tongue carcinomas tissues were detected by mmunohistochemical method. The expression of Shp2 in Cal-27 cell was inhibited by short interfering RNA interference （siRNA）. The proliferation of Cal-27 cells at 24 h, 48 h, 72 h, 96 h were detected by MTT. The apoptosis of Cal-27 cells influenced by Shp2 were detected by flow cytometry. The expres- sions of Bax, p53, Bcl-2 proteins in Cal-27 cells were detected by western blot. The invasion and metastasis of Cal-27 cells influenced by Shp2 were detected by transwell assay and wound healing assay. Result The positive expression rate of Shp2 in tongue carcinomas tissues was significantly higher than that in the normal adjacent tissues （P〈0.01）. The pro- liferation, invasion and metastasis abilities of Shp2-siRNA Cal-27 cells were significant lower than those of Cal-27 cells （P〈0.05）. Compared to blank control, the apoptosis rates of Shp2-siRNA Cal-27 cells and Cal-27 cells were both signifi- cantly increased （P〈0.05）. The expression levels of Bax, p53 protein in Shp2-siRNA Cal-27 cells were significantly high- er than those in blank control and negative control, and the expression level of Bcl-2 protein in Shp2-siRNA Cal-27 cells was significantly lower than that in blank control and negative control （P〈0.05）. Conclusion Shp2 has anticancer activi- ty in tongue carcinoma cells, which can promote the proliferation, metastasis and invasion of Cal-27 cells and inhibit their apoptosis.