hTRT催化功能区基因克隆及其在肿瘤中的表达

目的 研究端粒酶蛋白ｈＴＲＴ基因在肿瘤中的表达及其意义。方法 用ＲＴ－ＰＣＲ法检查Ｈｅｌａ细胞与ＰＧ细胞的ｈＴＲＴ表达水平,将Ｈｅｌａ细胞的ｈＴＲＴ催化功能克隆,并进行测序比较,应用原位杂交技术对肿瘤组织中的ｈＴＲＴ和ｈＴＲ（端粒酶ＲＮＡ组分）基因进行检测。结果 Ｈｅｌａ细胞与ＰＧ细胞均有ｈＴＲＴ表达,Ｈｅｌａ细胞ｈＴＲＴ催化功能区ｃＤＮＡ序列与文献报道一致,原位杂交结果显示ｈＴＲＴ与ｈＴＲ的协同

Molecular cloning of hTRT catalytic domain and its expression in tumors

Objective To study the expression of hTRT gene in tumors and its significance in cancer diagnosis. Methods The expression of hTRT in HeLa cells and PG cells was estimated by RT PCR and the hTRT cDNA encoding the catalytic domain from HeLa cells were cloned and sequenced. Using in situ hybridization (ISH) techniques, the expression of hTRT mRNA in 54 human tumors were observed and compared with the expression of hTR. Results hTRT was detected in both HeLa and PG cells. Sequencing showed that the cloned hTRT catalytic domain cDNA was identical with that reported in the literature. hTRT and hTR expressions were detected in 38/46 and 40/46 malignant tumors respectively, while both hTRT and hTR were not detected in 8 benign tumors. Conclusion hTRT was detected in both HeLa and PG cancer cells. hTRT catalytic domains were highly conservative. The expression of hTRT and hTR were both high in malignant tumors and they were strongly correlated. Our results suggest that detection of hTRT was valuable to clinical oncology diagnosis.