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作 者:顾青 赵艳 陈华 GU Qing ZHAO Yan CHEN Hua(Department of Obstetrics and Gynecology, Shanghai Baoshan Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai201999)
机构地区:[1]上海市宝山区中西医结合医院妇产科,上海201999
出 处:《实用中西医结合临床》2017年第6期1-4,56,共5页Practical Clinical Journal of Integrated Traditional Chinese and Western Medicine
摘 要:目的:探讨MTA1基因沉默对宫颈癌细胞转移增殖的影响。方法:用慢病毒转染法稳定转染Siha细胞,转染同时进行实验分组。用RT-PCR技术和蛋白印迹法测定宫颈癌Siha、Hela细胞中MTA1 m RNA和蛋白的表达。Transwell体外侵袭实验检测转染后Siha细胞的迁移能力,MTT检测法和克隆形成实验法检测转染后Siha细胞的细胞增殖能力,FACS细胞凋亡法检测转染后Siha细胞的细胞凋亡率,PI-FACS细胞周期法检测转染后Siha细胞的各个生长周期的细胞数。结果:(1)Transwell体外侵袭实验:相比NC组,KD组Transwell转移率经T-Test分析P=2.61E-08<0.05。(2)MTT检测结果表明:相比NC组,KD组细胞增殖减缓。(3)克隆形成实验结果显示:相比NC组,KD组克隆数经T-Test分析P值=0.000 6<0.05。(4)FACS细胞凋亡:相比两个对照组,KD组凋亡率经T-Test分析P<0.05。结论:MTA1基因促进宫颈癌细胞的转移和增殖,沉默MTA1基因表达能使宫颈癌细胞增殖及迁移能力下降,加速宫颈癌细胞的凋亡,为抑制肿瘤转移奠定实验基础,最后为宫颈癌的新型药物性靶向治疗提供有力的实验依据。Objective: To investigate the effects of MTA1 gene silencing on the metastatic and proliferation ability of cervical cancer cell. Methods: The experiment was divided into three groups when the Siha cells were infected by lentivirus. Reverse transcription(RT)-PCR and western blot were used to detect MTA1 m RNA and protein expressions in Siha cell and Hela cell of cervical cancer. Transwell assay detected the migration ability of transfected Siha cells. The proliferation ability of transfected Siha cells were determined by methyl thiazolyl tetrazolium(MTT) and clone formation experiment. FACS cell apoptosis method detected apoptosis rate of transfected Siha cell. Flow cytometry detected transfected Siha cells at different growth periods. Results:(1) Transwell assay: Compared with NC group, the transfer rate in the KD group were analyzed by T Test(P=2.61 〈0.05).(2)MTT test: Compared with NC group, the proliferation index of cells was significantly decreased in KD group.(3)Clone formation experiment: Compared with NC group, the numbers of cloning were significantly less than KD group(P=0.0006 〈0.05).(4)FACS: Compared with NC group and Mock group,the apoptosis rate was significantly lower than KD group(P〈 0.05). Conclusions: MTA1 gene may premote the metastasis and proliferation ability of cervical cancer cell.MTA1 gene silencing may decrease the proliferation and migration of cervical cancer cell, while accelerating the apoptosis of cervical cancer cell. It provides experiental basis for the inhibition of neoplasm metastasis, and powerful new experimental basis for cervical cancer drug targeted therapy finally.
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