PTD-BCR/ABL SH3融合蛋白对肝癌细胞株SMMC-7721的杀伤作用

In vitro Study on the Cytotoxicity of PTD-BCR/ABL SH3 Fusion Protein to Hepatocellular Cancer Cell Line SMMC-7721

目的:探讨PTD鄄BCR/ABLSH3融合蛋白对肝癌细胞株SMMC鄄7721的体外杀伤活性。方法:通过将PTD鄄BCR/ABLSH3融合蛋白与SMMC鄄7721细胞共培养,用免疫细胞化学染色的方法观察和检测PTD鄄BCR/ABLSH3融合蛋白进入细胞后的定位,MTT法检测蛋白对肝癌细胞的杀伤活性,电镜观察SMMC鄄7721肝癌细胞与PTD鄄BCR/ABLSH3融合蛋白共培养后超微结构变化。结果:PTD鄄BCR/ABLSH3融合蛋白进入SMMC鄄7721细胞株后主要分布在细胞核,少量分布于胞浆。融合蛋白对SMMC鄄7721细胞株的杀伤作用呈浓度依赖性,在所选蛋白浓度和观察时限内,当蛋白浓度≥0.14mg/ml、共培养4h后,SMMC鄄7721细胞形态就出现改变,8h后出现死亡细胞。电镜显示,PTD鄄BCR/ABLSH3融合蛋白促使SMMC鄄7721细胞株凋亡。结论:PTD鄄BCR/ABLSH3融合蛋白能抑制SMMC鄄7721细胞株增殖,促进其凋亡,并改变其骨架结构。

Objective: To study the cytotoxicity of PTD-BCR/ABL SH3 fusion protein to hepatocellular cancer cell line SMMC-7721 in vitro. Methods: SMMC-7721 cells were cultured with the addition of PTD-BCR/ABL SH3 fusion protein. The localization of PTD-BCR/ABL SH3 fusion protein was detected by immunocytochemical staining. The cytotoxicity was measured with the method of MTT. Electronic microscopy was used to observe the changes in ultrastructure of SMMC-7721 cells after culturing with the addition of PTD-BCR/ABL SH3 fusion protein. Results: Most of PTD-BCR/ABL SH3 fusion protein was found in cell nuclei and little in plasma. The cytotoxitity caused by PTD-BCR/ABL SH3 fusion protein was dose-dependent. When the concentration of PTD-BCR/ABL SH3 fusion protein exceeded 0.14 mg/ml, and after 4 hours of culture, the SMMC-7721 cells displayed abnormal morphology, and after 8 hours, the cells died. Electronic microscopic photos showed that PTD-BCR/ABL SH3 fusion protein induced obvious characters of apoptosis in the SMMC-7721 cells. Conclusions: PTD-BCR/ABL SH3 fusion protein restrics the proliferation of hepatocellular cancer cells and enhances their apoptosis, thus alters the structural framework of the cancer cells.