PCNA、Survivin蛋白在人绒癌细胞株BeWo合体化过程中表达变化的研究

Expressions of PCNA and Survivin Proteins in Be Wo Cells during Syncytial Process

目的:通过检测人绒癌细胞株Be Wo合体化过程中增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、生存素(Survivin)蛋白表达的变化,探讨滋养细胞合体化后增殖性的变化,为恶性滋养细胞肿瘤,尤其是耐药恶性滋养细胞肿瘤的临床治疗提供新的思路和方法。方法:利用毛喉素(forskolin)诱导Be Wo细胞株融合;应用逆转录聚合酶链反应(RT-PCR)检测促融素(Syncytin)在forskolin作用不同时间的Be Wo细胞株中的表达;应用蛋白质印迹(Western blotting)检测PCNA、Survivin蛋白在forskolin作用不同时间的Be Wo细胞株中的表达;应用噻唑蓝比色分析实验(MTT)法检测forskolin作用不同时间的绒癌细胞株Be Wo的增殖能力。结果:1forskolin作用后的Be Wo细胞株Syncytin基因的表达增强,且随着forskolin作用时间的延长,Syncytin的表达更强,于48 h达到高峰。2forskolin作用后的Be Wo细胞株PCNA、Survivin蛋白的表达降低。3forskolin作用后的Be Wo细胞株的增殖能力下降,且不同作用时间的差异有统计学意义;forskolin作用的时间越长,Be Wo细胞株增殖能力下降越明显。结论:人绒癌细胞株Be Wo合体化后PCNA、Survivin蛋白的表达降低,说明人绒癌细胞株Be Wo合体化后增殖性降低,推测诱导滋养细胞合体化可能对临床治疗恶性滋养细胞肿瘤具有一定作用。

Objective：To observe the expression changes of PCNA and Survivin proteins in the choriocarcinoma cell line（Be Wo） during its fusion, so as to explore the proliferation change after the fusion of trophoblast cells. The study could be helpful for developing a new therapy of gestational trophoblastic tumor（GTT）, especially drug-resistant GTT. Methods：The intercellular fusion of the cultured Be Wo cells was induced by forskolin. The expression of Syncytin m RNA in the treated Be Wo cells was detected by RT-PCR, and the expressions of PCNA and Survivin proteins were detected by Western blotting. The proliferation of Be Wo cells was detected by MTT. Results： 1 The expression of Syncytin gene in the Be Wo cell treated with forskolin was increased with a time-dependent manner, which reached its peak in 48 hours. 2 The expressions of PCNA and Survivin proteins in Be Wo cell after treatment were significantly decreased. 3The proliferation of the Be Wo cell treated with forskolin was significantly decreased with a time-dependent manner. Conclusions：The lowered expressions of PCNA and Survivin proteins in those fusion choriocarcinoma cells means the lowered proliferation,suggesting that the induced intercellular fusion of trophoblast cells may be helpful to treat those malignant GTT.