摘要
目的为了探讨P53基因对腺样囊性癌细胞端粒酶活性的抑制作用及机制。方法构建携带人野生型P53基因的腺病毒表达载体,以脂质体法瞬时转染腺样囊性癌SACC83细胞,RT PCR检测转染细胞P53基因mRNA的表达,TRAP PCR ELISA法检测转染细胞端粒酶活性的改变。并将P53基因与含有hTERT启动子核心调控区的荧光素酶报告基因pGL2630共转染SACC83细胞,测定转染细胞荧光素酶报告基因活性。结果1.瞬时转染含人野生型P53基因的重组腺病毒表达载体pΔE1P53于腺样囊性癌SACC83细胞后,其P53基因mRNA的表达明显增强;2.基因转染后,SACC83细胞内源性端粒酶活性显著降低。3.P53基因与含有hTERT启动子核心调控区的荧光素酶报告基因pGL2630共转染SACC83细胞后,其荧光素酶活性显著下降。结论外源性表达P53基因可以降低腺样囊性癌细胞SACC83细胞端粒酶活性,并可能通过抑制端粒酶hTERT基因启动子的转录活性实现。
Objective To investigate the effect of P53 in telomerase activity in salivary adenoid cystic carcinoma cells. Human wild-type P53 gene was constructed into pΔE1 vector. Methods Liposome-mediated gene transfection method was used to transfer P53 gene into SACC-83 cells. The expression of P53 mRNA was assessed by reverse transcription polymerase chain reaction(RT-PCR) and the activity of telomerase was examined by TRAP-PCR-ELISA. pΔE1-P53 and hTERT promoter-luciferase reporter vector pGL2-630 were co-transfected into SACC-83 cells, and then the luciferase activity was examined. Results 1. The level of P53 mRNA increased after pΔE1-P53 transfection; 2. The activity of telomerase decreased obviously. 3. The luciferase activity was also depressed after the co-transfection of pΔE1-P53 and pGL2-630. Conclusion The results suggested that P53 gene may depress the activity of telomerase in salivary adenoid cystic carcinoma cell through inhibiting the transcriptional activity of hTERT promoter.
出处
《基础医学与临床》
CSCD
北大核心
2005年第3期204-208,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(30271422)
河北省自然科学基金(C2004000624)
国家留学人员科技活动择优资助项目
