目的：研究长链非编码RNA（long non-coding RNA，lncRNA）CCAT2 对宫颈癌细胞增殖、周期的影响。方法：采用qPCR法检测3 种宫颈癌细胞（HeLa、C-33A和CaSki）中CCAT2 的表达水平，选择表达水平最高的细胞进行后续实验。设计合成CCAT2 过表达及干扰载体，转染细胞后，qPCR检测转染效率。细胞分为5 组：对照组、干扰空载（sh-EV）组、过表达空载（overExp-EV）组、CCAT2 干扰（sh-CCAT2）组和CCAT2 过表达（overExp-CCAT2）组，MTT法检测各组细胞增殖活力，流式细胞术检测细胞周期，WB法检测细胞中Ki67、细胞周期蛋白D1（Cyclin D1）和周期蛋白依赖性激酶4（cyclin-dependent kinase 4，CDK4）表达水平。结果：在HeLa、C-33A和CaSki 3 种细胞系中，选择CCAT2 表达水平最高的CaSki 细胞为研究对象。CCAT2 过表达及干扰载体均成功转入细胞，转染效果明显。与对照组比较，sh-CCAT2 组细胞增殖活力显著降低（P<0.01）、G1 期细胞比例显著升高（P<0.01），Ki67、Cyclin D1 及CDK4 表达水平显著降低（均P<0.01）；而overExp-CCAT2 组与之相反，细胞增殖能力增强，且Ki67、Cyclin D1 及CDK4 表达水平显著升高（均P<0.01）。结论：CCAT2 通过调控细胞增殖及周期相关蛋白的表达，进而影响宫颈癌CaSki细胞的增殖和周期。

Objective: To investigate the effect of long non-coding RNA (lncRNA)-CCAT2 on the proliferation and cell cycle of cervical cancer cells. Methods: The expression of CCAT2 in 3 cervical cancer cell lines (HeLa, C-33A, and CaSki) was detected by qPCR and the cell line with the highest expression level was selected for subsequent experiments. CCAT2 overexpression and interference vectors were designed and synthesized. After transfection, qPCR was performed to detect the transfection efficiency. The cells were divided into 5 groups: control, sh-EV (empty vector), overExp-EV, sh-CCAT2, and overExp-CCAT2. MTT assay was performed to evaluate cell viability. Flow cytometry was performed to measure cell cycle. WB was performed to detect the expressions of Ki67, cyclin D1, and cyclin dependent kinase 4 (CDK4). Results: Among HeLa, C-33A, and CaSki cells, the highest expression of CCAT2 was found in CaSki cells. CCAT2 overexpression and interference vectors were successfully transfected into the CaSki cells.Compared with the control group, the cells viability and proliferation in the sh-CCAT2 group was significantly decreased (all P<0.01),the proportion of cells in the G1 phase was significantly increased (P<0.01), and the expression levels of Ki67, cyclin D1, and CDK4 were significantly decreased (all P<0.01). However, in the overExp-CCAT2 group, the cell proliferation was enhanced and the expression levels of Ki67, cyclin D1, and CDK4 were significantly increased (all P<0.01). Conclusion: CCAT2 affects proliferation and cell cycle of cervical cancer cells by regulating the expressions of their associated proteins.

湖北省卫生健康委员会2019 年面上资助项目（No.WJ2019M073）