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目的探讨卷曲螺旋结构域蛋白34(CCDC34)在宫颈癌细胞中的表达及其对宫颈癌细胞增殖、凋亡、迁移和侵袭力的影响。方法收集南方医科大学第三附属医院妇产科2014—2016年间的16例宫颈鳞癌标本和16例慢性宫颈炎组织标本。荧光定量RT-PCR及Western Blot检测宫颈癌组织和细胞中CCDC34的表达;通过构建CCDC34慢病毒载体干扰CCDC34的表达,荧光定量RT-PCR和Western blot验证CCDC34的干扰效果。使用噻唑蓝(MTT)法、Brd U掺入实验、Transwell小室侵袭实验、流式细胞术来检测干扰CCDC34对Hela细胞增殖、迁移、侵袭能力及凋亡的影响。应用Caspase-3活性检测试剂盒及Western blot检测沉默CCDC34后Caspase-3的活性变化及在蛋白水平的表达。结果 CCDC34在宫颈癌细胞及组织中高表达;MTT及Brd U掺入实验显示,CCDC34干扰组的细胞增殖能力较对照组显著降低(P<0.05);Transwell迁移与侵袭实验显示,CCDC34干扰组穿膜细胞数与对照组相比显著减少[(57±7.9)个/200倍视野vs.(98±9.2)个/200倍视野,P<0.05;(33±5.2)个/200倍视野vs.(70±5.7)个/200倍视野,P<0.05],细胞凋亡显著增加[(27.60±5.30)%vs.(9.45±1.90)%]及Caspase-3的活性明显增强。结论沉默CCDC34的表达可显著抑制宫颈癌细胞的增殖、迁移和侵袭,促进细胞凋亡。
Objective To investigate the expression of coiled coil domain protein 34 (CCDC34) in cervical cancer cells and its effect on the proliferation, apoptosis, migration and invasiveness of cervical cancer cells. Methods 16 cases of cervical squamous cell carcinoma and 16 cases of chronic cervicitis from 2014 to 2016 were collected from the Third Affiliated Hospital of Southern Medical University. The expression of CCDC34 in cervical cancer tissues and cells was detected by fluorescent quantitative RT-PCR and Western Blot. CCDC34 expression was detected by constructing CCDC34 lentiviral vector. The interference effect of CCDC34 was confirmed by quantitative RT-PCR and Western blot. The effects of interfering CCDC34 on the proliferation, migration, invasion and apoptosis of Hela cells were detected by MTT assay, Brd U incorporation assay, Transwell chamber invasion assay and flow cytometry. Caspase-3 activity assay kit and Western blot were used to detect the changes of Caspase-3 activity and protein expression after silencing CCDC34. Results CCDC34 was highly expressed in cervical cancer cells and tissues. MTT and BrdU incorporation assay showed that the cell proliferation ability of CCDC34 interference group was significantly lower than that of the control group (P <0.05). Transwell migration and invasion assay showed that CCDC34 interference group penetrated the membrane (57 ± 7.9) / 200 times field of view vs. (98 ± 9.2) / 200 times field of view, P <0.05; (33 ± 5.2) / 200 times field vs. ( 70 ± 5.7) cells / 200 times visual field, P <0.05]. The apoptosis of cells was significantly increased [(27.60 ± 5.30)% vs (9.45 ± 1.90)%] and Caspase-3 activity. Conclusion Silencing CCDC34 expression can significantly inhibit the proliferation, migration and invasion of cervical cancer cells and promote apoptosis.