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目的 探讨端粒酶反义寡核苷酸对人乳腺癌细胞端粒酶活性的影响及抑制端粒酶活性以控制乳腺癌细胞生长的可能性。方法 采用 10 μmol/L与端粒酶催化亚基 (hTRT )mRNA互补的反义寡核苷酸处理乳腺癌细胞 ;于处理后 2 4、72h用端粒重复序列扩增及酶联免疫吸附方法检测细胞端粒酶活性 ;于处理后 72h以流式细胞术测定细胞周期 ;于处理后 12 0h采用原位末端标记法检测细胞凋亡。结果 经hTRT反义寡核苷酸作用后 ,细胞端粒酶活性明显受到抑制 ,至作用后 72h ,端粒酶活性较对照组降低 65 .6% (P <0 .0 5 ) ;细胞生长明显受到抑制 ;细胞周期发生显著变化 :G0 /G1期细胞数增多 ,S期及G2 /M期细胞数则减少 ;细胞增殖指数由 0 .46降低至 0 .2 5 ;并可观察到凋亡细胞的出现 ,凋亡发生率为 12 .5 % ;而无义组及空白对照组以上各指标均无明显变化 (P >0 .0 5 )。结论 端粒酶反义寡核苷酸可明显抑制乳腺癌细胞端粒酶活性 ,抑制细胞生长和增殖 ,诱导细胞凋亡 ;表明应用反义技术抑制端粒酶活性治疗乳腺癌具有广阔的前景。
Objective To investigate the effect of telomerase antisense oligonucleotide on telomerase activity of human breast cancer cells and the possibility of inhibiting telomerase activity to control the growth of breast cancer cells. Methods Human breast cancer cells were treated with 10 μmol / L antisense oligodeoxynucleotides complementary to telomerase catalytic subunit (hTRT) mRNA. Twenty-four hours and 72 hours after treatment, telomeric repeat amplification and enzyme-linked immunosorbent assay Cell telomerase activity was measured. Cell cycle was determined by flow cytometry at 72h after treatment. Apoptosis was detected by in situ terminal labeling method at 12h after treatment. Results After treated with hTRT antisense oligonucleotide, the telomerase activity was significantly inhibited, and the telomerase activity decreased by 65.6% (P <0. 05) Was inhibited; the cell cycle changed significantly: the number of cells in G0 / G1 phase increased, the number of cells in S phase and G2 / M phase decreased; the cell proliferation index decreased from 0.46 to 0.25; and apoptotic cells were observed The incidence of apoptosis was 12.5%, while there was no significant difference in the above indexes between the non-sense group and the blank control group (P> 0.05). CONCLUSION: Antisense oligonucleotide of telomerase can significantly inhibit telomerase activity, inhibit cell growth and proliferation and induce apoptosis in breast cancer cells. It is indicated that antisense technology can inhibit telomerase activity in the treatment of breast cancer.