富含半胱氨酸蛋白61对子宫内膜癌细胞增殖及细胞周期的影响及其机制研究

Effect of cysteine-rich protein 61 on proliferation and cell cycle of endometrial cancer cells and its mechanism

摘要:

目的 研究富含半胱氨酸蛋白61（CYR61）对子宫内膜癌（EC）细胞株HEC-1B增殖及细胞周期的影响，并探讨其机制。方法 取对数生长期HEC-1B细胞，随机分为Control组（不处理）、pcDNA3.1-NC组（转染pcDNA3.1-NC）、pcDNA3.1 CYR61组（转染pcDNA3.1 CYR61）、pcDNA3.1 CYR61+ LiCl （Wnt/β-catenin通路激活剂）组（转染pcDNA3.1 CYR61，并加入LiCl使其终浓度为60 mmol/L）。转染48 h后采用MTT法检测增殖抑制率，PI染色法检测细胞周期占比，Western blotting检测细胞β-catenin、Caspase-3、Cyclin D1蛋白的表达。结果 与Control组、pcDNA3.1-NC组比较，pcDNA3.1 CYR61组CYR61、β-catenin、Caspase-3、Cyclin D1蛋白相对表达量、G₀/G₁期占比升高（P <0.05），S、G₂/M期占比降低（P <0.05）；与pcDNA3.1 CYR61组比较，pcDNA3.1 CYR61+ LiCl组CYR61、β-catenin、Caspase-3、Cyclin D1蛋白相对表达量、G₀/G₁期占比降低（P <0.05），S、G₂/M期占比升高（P <0.05）。4组24 h、48 h、72 h增殖抑制率比较，采用重复测量设计的方差分析，结果 ①不同时间点增殖抑制率有差异（F =25.416，P =0.000）；②3组增殖抑制率有差异（F =46.962，P =0.000），pcDNA3.1 CYR61组增殖抑制率较高，增殖抑制效果好；③3组增殖抑制率变化趋势有差异（F =37.541，P =0.000）。结论 上调CYR61可抑制EC细胞株HEC-1B增殖，阻滞其细胞周期，其作用机制可能与抑制Wnt/β-catenin信号通路有关。

Abstract:

Objective To study the effect of cysteine-rich protein 61 (CYR61) on the proliferation and cell cycle of endometrial cancer (EC) cell line HEC-1B, and to explore the possible mechanism.Methods Log phase HEC-1B cells were taken and randomly divided into control group (no treatment), pcDNA3.1-NC group (transfected with pcDNA3.1-NC), pcDNA3.1 CYR61 group (transfected with pcDNA3.1 CYR61), pcDNA3.1 CYR61 + LiCl [Wnt/β-catenin (β-catenin) pathway activator] group (pcDNA3.1 CYR61 was transfected, and LiCl was added to make the final concentration 60 mmol/L). At 48h after transfection, the proliferation inhibition rate was detected by MTT experiment. The cycle proportion was detected by PI staining method. The protein expression level of β-catenin, aspartate-specific caspase-3 (Caspase-3), and Cyclin D1 (Cyclin D1) in different groups were detected by Western blotting method.Results Compared with control group and pcDNA3.1-NC group, the proportion of S, G₂/M phase, and the protein expression of β-catenin, Caspase-3, Cyclin D1 in the pcDNA3.1 CYR61 group were decreased, and the CYR61 protein expression and the proportion of G₀/G₁ phase was increased (P < 0.05). Compared with pcDNA3.1 CYR61 group, the proportion of S, G₂/M phase, and the protein expression of β-catenin, Caspase-3, Cyclin D1 in the pcDNA3.1 CYR61 + LiCl group were increased, and the CYR61 protein expression, the proportion of G₀/G₁ phase was decreased (P < 0.05). The proliferation inhibition rates of pcDNA3.1-NC group, pcDNA3.1 CYR61 group, and pcDNA3.1 CYR61 + LiCl group were different (F = 46.962, P = 0.000). The pcDNA3.1 CYR61 group had higher proliferation inhibition rate and better proliferation inhibition effect. There were differences in the change trend of proliferation inhibition rate among pcDNA3.1-NC group, pcDNA3.1 CYR61 group, and pcDNA3.1 CYR61 + LiCl group (F = 37.541, P = 0.000).Conclusion Up-regulation of CYR61 can inhibit the proliferation of EC cell line HEC-1B and block its cell cycle, which may be achieved by inhibiting the Wnt/β-catenin signaling pathway.