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目的探讨煤焦沥青致人支气管上皮细胞(BEAS-2B)恶变细胞的端粒损伤作用。方法用中温煤焦沥青烟提取物作为诱导剂,建立永生化BEAS-2B的恶性转化模型,用实时荧光定量聚合酶链反应(RT-PCR)法检测端粒DNA长度和端粒酶活力变化。结果煤焦沥青烟提取物诱导BEAS-2B细胞后,30代时诱导组细胞能在软琼脂上形成典型的阳性克隆,染色体二倍体核型比例显著降低,形成恶性转化细胞。BEAS-2B恶性转化细胞端粒DNA长度缩短(0.41±0.03),与正常对照组(1.01±0.08)和二甲基亚砜(DMSO)组(0.92±0.04)比较,差异均有统计学意义(P<0.001)。BEAS-2B恶性转化细胞端粒酶活力增强(1.62±0.07),与正常对照组(1.03±0.08)和DMSO组(1.11±0.03)比较,差异均有统计学意义(P<0.001)。结论端粒损伤在煤焦沥青致细胞恶性转化中起重要作用。
Objective To investigate the telomere injury induced by coal tar pitch in human malignant bronchial epithelial cells (BEAS-2B). Methods The malignant transformation model of immortalized BEAS-2B was established by using medium-temperature coal tar pitch smoke as an inducer, and the changes of telomere length and telomerase activity were detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR). Results After BEAS-2B cells were induced by coal tar pitch smoke extract, the typical positive clones were formed on the soft agar in the 30th generation. The karyotype of the diploid chromosomes was significantly reduced and the malignant transformed cells formed. The length of telomere DNA was shortened in BEAS-2B malignant transformed cells (0.41 ± 0.03), which was significantly different from that in normal control group (1.01 ± 0.08) and DMSO group (0.92 ± 0.04) P <0.001). The telomerase activity of BEAS-2B malignant transformed cells was enhanced (1.62 ± 0.07), which was significantly different from that of normal control group (1.03 ± 0.08) and DMSO group (1.11 ± 0.03) (P <0.001). Conclusion Telomere injury plays an important role in the malignant transformation of coal tar pitch-induced cells.