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作 者:卢星轩 张艳[1] 曾益军[1] 郑英如[2] 刘东擘[1] 何凤田[1]
机构地区:[1]第三军医大学基础医学部生物化学与分子生物学教研室,重庆400038 [2]第三军医大学大坪医院野战外科研究所妇产科,重庆400042
出 处:《第三军医大学学报》2016年第4期361-366,共6页Journal of Third Military Medical University
基 金:国家自然科学基金面上项目(81272865);国家自然科学基金青年科学基金(81402268);重庆市自科学基金(CSTC2013jj B10015)~~
摘 要:目的探讨过表达SARI(suppressor of AP-1 and regulated by IFN)对宫颈癌细胞增殖的影响及可能的分子机制。方法在He La细胞和Ca Ski细胞转染可表达SARI的质粒后,使用Real-time PCR和Western blot检测细胞中SARI表达水平的变化;CCK-8法检测上述宫颈癌细胞模型的增殖;流式细胞术分析上述细胞模型的周期及凋亡;报告基因检测转录因子AP-1的转录活性;Western blot检测抗凋亡分子Mcl-1及Bcl-2的表达。结果成功构建过表达SARI的细胞模型。SARI表达水平上调后,细胞增殖受到抑制(P<0.01);早期凋亡细胞增加但细胞周期无明显变化。转录因子AP-1的转录活性及Mcl-1的表达明显下调(P<0.05),但Bcl-2的表达无明显变化。结论宫颈癌细胞中过表达SARI诱导宫颈癌细胞发生早期凋亡,可能是通过抑制AP-1的转录并下调抗凋亡分子Mcl-1的表达来实现的。Objective To investigate the effect of overexpression of suppressor of AP-1 and regulated by IFN (SARI) on the proliferation of cervical cancer cells and explore the underlying molecular mechanism. Methods HeLa cells and CaSki cells were transformed with plasmids capable of expressing SARI. The expression of SARI was detected by real-time PCR and Western blotting. Meanwhile, the proliferation of the transformed cancer cells was assessed by CCK-8 assay, and cell cycle and apoptosis were detected by flow cytometry. In addition, the transcriptional activity of AP-1 transcription factor was determined by luciferase reporter assay, and the expression of Mcl-1 and Bcl-2 was analyzed by Western blotting. Results A cell model of over-expressing SARI was successfully established (P 〈 0. 01 ). After up-regulating SARI, the cell proliferation was inhibited, early apoptotic cells were increased, but no significant change was observed in the cell cycle. Moreover, the transcriptional activity of AP-1 and the expression of Mcl-1 were both significantly suppressed (P 〈 0. 05), while there was no change in the expression of Bel-2. Conclusion Overexpression of SARI can induce early apoptosis of cervical cancer cells through suppressing the transcriptional activity of AP-1 and down-regulating the expression of Mcl-1.
关 键 词:SARI 宫颈癌 细胞增殖 AP-1 MCL-1
分 类 号:R394.3[医药卫生—医学遗传学] R730.23[医药卫生—基础医学]
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