论文部分内容阅读
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(P<0.05)。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)%,P<0.05)];(2)G0/G1期细胞百分率显著增加[(82.56±3.89)%vs(61.37±3.28)%,P<0.05]、S期细胞百分率显著减少[(9.12±2.48)%vs(28.97±2.08)%,P<0.01];(3)细胞早期凋亡率和晚期凋亡率均显著增加(均P<0.05);(4)细胞迁移率显著降低(P<0.01);(5)Rho A和ROCK 1蛋白表达水平均显著下降(均P<0.01)。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。
Objective: To investigate the effect of Klotho gene on the biological characteristics of human cervical squamous cell carcinoma Si Ha and its mechanism. Methods: The expression of Klotho gene was detected by real-time fluorescence quantitative PCR and Western blotting in Si Ha, He La and C33A cells and immortalized human epidermal HaCa T cells. The Klotho overexpression vector was constructed and transfected into Si Ha cells. The changes of proliferation, apoptosis and migration of Si Ha cells were detected by Transwell invasion assay and flow cytometry (CCK-8). The protein expressions of Rho A and ROCK 1 were detected by Western blotting Level. Results: The expression of Klotho in Si ha, Hela and C33A cells was lower than that in HaCa T cells (P <0.05). The proliferation of Klotho in Si Ha cells was significantly lower than that in the control group [(67.37 ± 5.04)% vs (100.34 ± 7.62)%, P <0.05)]; (2) The percentage of cells in S phase was significantly decreased ([(82.56 ± 3.89)% vs (61.37 ± 3.28)%, P <0.05], and the percentage of cells in S phase was significantly decreased [(9.12 ± 2.48)% vs (28.97 ± 2.08)%, P <0.01] (3) The rates of early apoptosis and late apoptosis were significantly increased (all P <0.05); (4) the cell migration rate was significantly decreased (P <0.01); (5) The expression of Rho A and ROCK 1 was significantly Decreased (all P <0.01). Conclusion: Klotho can inhibit the proliferation and migration of Si ha cells and promote the apoptosis of human cervical cancer cells. The mechanism may be related to the inhibition of the activation of Rho A / ROCK 1 signaling pathway, which suggests that Klotho can be used as a diagnostic and targeted therapy for cervical Potential sites of cancer.