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目的研究端粒酶在宫颈癌及其癌前病变组织中的表达,探讨人端粒酶逆转录酶(hTERT)作为宫颈癌早期诊断标志物的可能性。方法采用RTPCR方法测定端粒酶mRNA在3株宫颈癌细胞、2株正常的人二倍体细胞、38例宫颈癌、16例宫颈上皮内瘤变(CIN)和20例正常宫颈组织中的表达水平;端粒重复序列扩增酶联免疫吸附法测定端粒酶活性;免疫组化链酶菌抗生物素蛋白过氧化物酶(SP)连接法检测3株宫颈癌细胞、2株正常的人二倍体细胞、21例正常宫颈组织、2例CIN和55例宫颈癌切片中hTERT蛋白的表达水平。结果hTERTmRNA在3株宫颈癌细胞中均表达,而在2株正常的人二倍体细胞中无表达,宫颈癌组织中的阳性表达率为81.6%,高于CIN的37.5%和正常宫颈组织的5.0%,差异有统计学意义(P<0.05);hTERTmRNA的阳性表达率与端粒酶活性有相关性(P<0.01);hTERT蛋白在3株宫颈癌细胞中有表达,2株正常的人二倍体细胞中无表达,正常宫颈组织中阳性表达率为4.8%、CIN为28.0%,均低于宫颈癌组织的65.5%,差异有统计学意义(P<0.05)。结论宫颈癌组织和细胞中hTERTmRNA和蛋白阳性表达率均高于正常宫颈组织,其有可能作为宫颈癌早期诊断的标志物。
Objective To study the expression of telomerase in cervical cancer and its precancerous lesions and to explore the possibility of hTERT as an early diagnostic marker of cervical cancer. Methods The expression of telomerase mRNA in 3 cervical cancer cells, 2 normal human diploid cells, 38 cervical carcinoma, 16 cervical intraepithelial neoplasia (CIN) and 20 normal cervical tissues was detected by RTPCR method. The level of telomerase activity was detected by telomere repeat amplification enzyme-linked immunosorbent assay (ELISA). Three cervical cancer cell lines were detected by immunohistochemical streptavidin biotin peroxidase (SP) Diploid cells, 21 cases of normal cervical tissue, 2 cases of CIN and 55 cases of cervical cancer hTERT protein expression levels. Results hTERT mRNA was expressed in all three cervical cancer cells but not in two normal human diploid cells. The positive rate of hTERT mRNA in cervical cancer was 81.6%, higher than that of CIN (37.5%) and normal cervical tissue (P <0.05). The positive expression rate of hTERT mRNA was correlated with the telomerase activity (P <0.01). The expression of hTERT protein in 3 cervical cancer cells and 2 normal human There was no expression in diploid cells. The positive expression rate in normal cervical tissues was 4.8% and the CIN was 28.0%, both of which were lower than that of 65.5% of cervical cancer tissues. The difference was statistically significant (P <0.05). Conclusion The positive rate of hTERT mRNA and protein expression in cervical cancer tissues and cells is higher than that in normal cervical tissues, which may be used as a marker for early diagnosis of cervical cancer.