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目的探讨人类大肠癌细胞胞膜蛋白组织因子(TF)对大肠癌细胞抗凋亡作用的影响。方法提取人大肠癌Lovo细胞DNA,以梯度浓度的TF激活剂凝血因子VIIa(FVIIa)激活去血清培养Lovo细胞,以有效浓度TF抗体封闭阻断这一作用。用流式细胞术测定细胞凋亡率,以Western blot检测凋亡相关蛋白Caspase-3及Bcl-2的表达。结果 FVIIa能使细胞产生有效抵抗去血清培养诱导的细胞凋亡,其抗凋亡作用与FVIIa浓度呈现剂量效应关系。胞膜TF封闭抗体能有效阻断这一作用。Caspase-3及磷酸化Bcl-2蛋白在TF上述抗凋亡作用中被活化。结论激活大肠癌Lovo细胞TF能有效抑制细胞凋亡,且这一作用可被TF封闭抗体有效阻断。
Objective To investigate the effect of membrane protein and tissue factor (TF) on the anti-apoptotic effect of human colorectal cancer cells. Methods The DNA of human colorectal cancer cell line Lovo was extracted and the serum was cultured in Lovo cells with gradient activator of coagulation factor Vlla (FVIIa). Blockade of this effect was blocked with effective concentration of TF antibody. The apoptotic rate was determined by flow cytometry and the expression of Caspase-3 and Bcl-2 were detected by Western blot. Results FVIIa could effectively inhibit cell apoptosis induced by serum-free culture. The anti-apoptotic effect of FVIIa showed a dose-response relationship with FVIIa concentration. Membrane TF blocking antibodies can effectively block this effect. Caspase-3 and phosphorylated Bcl-2 proteins are activated in the above anti-apoptotic effect of TF. Conclusion Activation of TF in colorectal cancer Lovo cells can effectively inhibit apoptosis, and this effect can be effectively blocked by TF blocking antibody.