Klotho基因对子宫颈鳞癌细胞恶性生物学行为的影响及相关机制

发布时间：2018-08-26 20:35

【摘要】：目的:探讨Klotho基因对人子宫颈鳞癌细胞株Si Ha生物学特性的影响及其作用机制。方法:实时荧光定量PCR及Western blotting检测Klotho基因在子宫颈鳞癌细胞系Si Ha、He La和C33A及人永生化表皮细胞Ha Ca T细胞中的表达水平。构建Klotho过表达载体并转染Si Ha细胞,采用CCK-8法、Transwell侵袭实验和流式细胞术检测Si Ha细胞增殖、凋亡和迁移能力的变化;Western blotting检测Rho A和ROCK 1蛋白表达水平。结果:Klotho在子宫颈癌Si Ha、Hela、C33A细胞中表达水平低于Ha Ca T细胞(P0.05)。在Si Ha细胞中过表达Klotho后,(1)细胞的增殖能力显著低于对照组[(67.37±5.04)%vs(100.34±7.62)%,P0.05)];(2)G0/G1期细胞百分率显著增加[(82.56±3.89)%vs(61.37±3.28)%,P0.05]、S期细胞百分率显著减少[(9.12±2.48)%vs(28.97±2.08)%,P0.01];(3)细胞早期凋亡率和晚期凋亡率均显著增加(均P0.05);(4)细胞迁移率显著降低(P0.01);(5)Rho A和ROCK 1蛋白表达水平均显著下降(均P0.01)。结论:Klotho能够抑制人子宫颈癌Si Ha细胞的增殖和迁移,并促进其凋亡;作用机制可能与抑制Rho A/ROCK 1信号通路的活化有关,提示Klotho可以作为诊断和靶向治疗子宫颈癌的潜在作用位点。
[Abstract]:Objective: to investigate the effect of Klotho gene on the biological characteristics of human cervical squamous cell carcinoma cell line Si Ha and its mechanism. Methods: Real-time quantitative PCR and Western blotting were used to detect the expression of Klotho gene in cervical squamous cell carcinoma cell lines Si Ha,He La and C33A and in human immortalized epidermal cell line Ha Ca T. Klotho overexpression vector was constructed and transfected into Si Ha cells. The expression levels of Rho A and ROCK 1 protein were detected by CCK-8 assay and flow cytometry. The changes of cell proliferation, apoptosis and migration ability were detected by Western blotting. Results the expression of Klotho in Si Ha,Hela,C33A cells was lower than that in Ha Ca T cells (P0.05). After overexpression of Klotho in Si Ha cells, (1) the proliferative ability of the cells was significantly lower than that of the control group [(67.37 卤5.04) vs (100.34 卤7.62) P 0.05]; (2) the percentage of G0/G1 phase cells significantly increased [(82.56 卤3.89) vs (61.37 卤3.28) P 0.05] the percentage of S phase cells significantly decreased [(9.12 卤2.48) vs (28.97 卤2.08) P 0.01]; (3) the rate of early apoptosis and late apoptosis increased significantly (all); (4). Cell migration was significantly decreased (P0.01); (5) Rho A and ROCK 1 protein expression levels were significantly decreased (P0.01). Conclusion: Klotho can inhibit the proliferation, migration and apoptosis of human cervical carcinoma Si Ha cells, and the mechanism may be related to the inhibition of the activation of Rho A/ROCK 1 signaling pathway. The results suggest that Klotho can be used as a potential site for the diagnosis and targeted treatment of cervical cancer.
【作者单位】：安阳市肿瘤医院内五科;郑州大学第一附属医院肿瘤科;
【分类号】：R737.33

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