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目的探讨端粒酶和端粒结合蛋白(TRF)在细胞凋亡过程中的作用机制。方法甲基噻唑四唑法(MTT)检测阿霉素对舌鳞癌Tca8113细胞的抑制作用。通过姬姆萨染色和流式细胞仪观察和检测5mg/L阿霉素诱导凋亡情况;利用酶联免疫吸附实验分析端粒酶活性;采用免疫组化和免疫荧光标记法观察和检测TRF表达及其水平。结果5mg/L阿霉素处理Tca8113细胞5d和7d后,出现明显凋亡;端粒酶活性降低呈时间依赖性;TRF在细胞核内表达,其表达水平无统计学意义(P>0.05)。结论阿霉素诱导Tca8113细胞凋亡可降低端粒酶活性,但不改变TRF表达水平。
Objective To investigate the mechanism of telomerase and telomere binding protein (TRF) in apoptosis. Methods Methyl thiazolyl tetrazolium assay (MTT) was used to detect the inhibitory effect of doxorubicin on Tca8113 cells. The apoptosis induced by 5 mg / L doxorubicin was observed and detected by Giemsa staining and flow cytometry. The telomerase activity was analyzed by enzyme-linked immunosorbent assay. The expression of TRF was detected by immunohistochemistry and immunofluorescence staining And its level. Results After treated with 5 mg / L doxorubicin for 5 days and 7 days, apoptosis of Tca8113 cells was observed. Telomerase activity decreased in a time-dependent manner. TRF expression in nucleus was not significant (P> 0.05). Conclusion Adriamycin induced apoptosis of Tca8113 cells decreased telomerase activity but did not change the expression of TRF.