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MASPIN在卵巢癌SKOV3细胞中表达调控机理的研究

The Regulation Mechanism of MASPIN Expression in Ovarian Carcinoma

  • 摘要: 目的 构建并瞬时转染P63MASPIN报告基因质粒,检测转染前后MASPIN启动子活性的改变,初步探讨在卵巢癌中P63MASPIN表达的调控作用。 方法 构建MASPIN报告基因质粒,并与P63质粒(TAP63、ΔNP63)共转染卵巢癌SKOV3细胞,荧光素酶测定转染后MASPIN启动子活性的变化情况;P63质粒体外瞬时转染卵巢癌SKOV3细胞,半定量逆转录聚合酶链反应(RT-PCR)方法检测转染后MASPIN mRNA表达水平的变化。 结果 成功构建MASPIN报告基因质粒,并与P63质粒(TAP63、ΔNP63) 共转染SKOV3细胞,荧光素酶报告基因法检测发现TAP63转染组MASPIN启动子活性高于空载体组和ΔNP63组(P<0.05),而后两组之间差异无统计学意义(P>0.05);P63质粒(TAP63、ΔNP63)瞬时转染SKOV3细胞, RT-PCR方法检测发现,TAP63转染组MASPIN mRNA表达高于未转染组、空载体组和ΔNP63组(P<0.05),而后3组之间差异无统计学意义(P>0.05)。 结论 TAP63能增强MASPIN启动子转录活性及MASPIN mRNA的表达,MASPIN可能是TAP63一个新的分子作用靶点。

     

    Abstract: Objective To study the regulation of P63 on expression of MASPIN in ovarian cancer by observe MASPIN promoter activity changes before and after transient transfection of constructed P63 and MASPIN reporter gene plasmids. Methods The MASPIN reporter plasmid, fused with luciferase reporter gene, was constructed and transfected into SKOV3 cells together with P63 (TAP63, ΔNP63) express plasmid transiently. The MASPIN promoter activity was determined in both the transfected cells and controlled ones by Luciferase Assays and the transcription of MASPIN mRNA of them was evaluated with semi quantitative RT-PCR. Results The MASPIN reporter plasmid was successfully constructed and transiently transfected into SKOV3 cells together with P63 (TAP63, ΔNP63) expression plasmid. The data showed among the tested P63 splice variants, TAP63 remarkably activated MASPIN promoter transactivation (P<0.05). No significant difference in the activity level of MASPIN promoter was detected in the SKOV3-vector and SKOV3-ΔNP63 cells (P>0.05). The level of MASPIN mRNA expression was notably enhanced in SKOV3-TAP63 cell after transient transfected with TAP63 express plasmid (P<0.05), but no significant difference among the SKOV3, SKOV3-vector and SKOV3-ΔNP63 cell(P>0.05)was detected. Conclusion TAP63 can activate the transcription activity of MASPIN promoter, as well as regulate the expression of MASPIN. Put all together, these results suggested that MASPIN is a new molecular target of P63.

     

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