Objective: To investigate the effect of cyclin A1 on the invasion, metastasis, and prognosis of hepatocellular carcinoma (HCC). Methods: Immunohistochemistry (IHC) was used to detect the expressional condition of cyclin A1 in HCC and paraffin-embedded non-tumor adjacent tissues. Kaplan-Meier method was used for the survival analysis of patients with HCC. Western blot (WB) was used to detect the expression of cyclin A1 in HCCLM3 and QGY-7703 cells. Scratch wound healing assay, transwell migration, and invasion assay were used to detect the effect of cyclin A1 overexpression on cell migration and invasion ability. WB was used to detect changes in the expression of matrix metalloproteinase (MMP) 2, MMP9, and vascular endothelial growth factor (VEGF) after overexpression of cyclin A1. Measurement data were compared using a t-test and analysis of variance. Count data was measured using χ (2) test and the Log-rank method was performed for survival analysis. Results: Cyclin A1 expression rates were higher in the tissues of HCC patients with recurrent metastasis than in the tissues of patients without recurrent metastasis (60.42% vs. 46.81%, χ (2) = 4.711, P < 0.05). The overall postoperative survival time (OS) and disease-free survival (DFS) were shorter in patients with high cyclin A1 expression than those with low cyclin A1 expression (45.9 months vs. 53.1 months; 42.9 months vs. 51.3 months, and P < 0.01). The postoperative OS and DFS were shorter in patients with high cyclin A1 expression and recurrent metastasis than those with low cyclin A1 expression without recurrent metastasis (31.7 months vs. 43.9 months; 18.0 months vs. 31.5 months, and P < 0.05). HCCLM3 and QGY-7703 cells were higher in the cyclin A1-pEX group than in the empty vector (vector) group (1.56 ± 0.06 vs. 0.18 ± 0.01, t = 18.75, P < 0.001; 1.31 ± 0.05 vs.0.37 ± 0.02, t = 15.17, P < 0.001). The migrated distances of HCCLM3 cells in the cyclin A1-pEX group and the vector group were (536.7 ± 14.5) μm and (327.3 ± 9.3) μm, t = 11.84, P < 0.05, respectively, while the migrated distances of QGY-7703 cells in the two groups were (916.7 ± 35.3) μm and (320.0 ± 20.8) μm, t = 13.54, P < 0.01. The migrated numbers of HCCLM3 cells in the cyclin A1-pEX group and vector group were (37.3 ± 2.4) and (7.0 ± 1.2), t = 12.67, P < 0.001, and the number of invasive cells was (73.7 ± 4.1) and (12.6 ± 1.5), t = 12.36, P < 0.001, respectively. The migrated numbers of QGY-7703 cells in the two groups were (153.3 ± 6.0) and (17.7 ± 3.7), t = 17.59, P < 0.001, and the number of invasive cells was (45.0 ± 2.9) and (9.3 ± 1.5), t = 10.66, P < 0.001, respectively. The expression levels of MMP2, MMP9, and VEGF in HCCLM3 and QGY-7703 cells were significantly higher in the cyclin A1-pEX group than those in the vector group (P < 0.05). Conclusion: Cyclin A1 plays an important role in HCC invasion and metastasis, but HCC patients with high cyclin A1 expression have a poor prognosis. Hence, cyclin A1 has high guiding significance for evaluating patient prognosis.
目的： 探讨细胞周期蛋白（Cyclin）A1对肝细胞癌（HCC）浸润转移及预后的影响。 方法： 免疫组织化学检测Cyclin A1在HCC及其相应癌旁非瘤石蜡组织中的表达情况；Kaplan-Meier法对HCC患者进行生存分析。蛋白质印迹法（WB）检测HCCLM3和QGY-7703细胞中Cyclin A1的表达。划痕愈合实验、Transwell迁移和侵袭实验检测Cyclin A1过表达对细胞迁移及侵袭能力的影响。WB检测Cyclin A1过表达后细胞基质金属蛋白酶（MMP）2、MMP9和血管内皮生长因子（VEGF）的表达变化。计量资料比较用t检验、方差分析；计数资料用χ(2)检验；Log-Rank法进行生存分析。 结果： 复发转移患者HCC组织中Cyclin A1的高表达率高于未复发转移患者（60.42%与46.81%，χ(2) = 4.711，P < 0.05）；Cyclin A1高表达的患者术后总体生存时间（OS）及无病生存时间（DFS）均短于低表达患者（45.9个月与53.1个月；42.9个月与51.3个月，P值均<0.01）；Cyclin A1高表达的术后复发转移患者术后OS及DFS也均短于低表达的患者（31.7个月与43.9个月；18.0个月与31.5个月，P值均<0.05）。HCCLM3和QGY-7703细胞在Cyclin A1-pEX组中Cyclin A1表达水平高于空白载体（Vector）组(1.56±0.06与0.18±0.01，t = 18.75，P < 0.001; 1.31±0.05与0.37±0.02, t = 15.17，P < 0.001)；HCCLM3细胞在Cyclin A1-pEX组与Vector组的迁移距离分别为（536.7±14.5）μm与（327.3±9.3）μm，t = 11.84，P < 0.05；QGY-7703细胞在2组的迁移距离分别为（916.7±35.3）μm与（320.0±20.8）μm，t = 13.54，P < 0.01。HCCLM3细胞在Cyclin A1-pEX组与Vector组的迁移个数分别为（37.3±2.4）个与（7.0±1.2）个，t = 12.67，P < 0.001；侵袭细胞数分别为（73.7±4.1）个与（12.6±1.5）个，t = 12.36，P < 0.001。QGY-7703细胞在2组的迁移个数分别为（153.3±6.0）个与（17.7±3.7）个，t = 17.59，P < 0.001；侵袭细胞数分别为（45.0±2.9）个与（9.3±1.5）个，t = 10.66，P < 0.001。HCCLM3和QGY-7703细胞在Cyclin A1-pEX组的MMP2、MMP9和VEGF的表达水平明显高于Vector组（P值均<0.05）。 结论： Cyclin A1在HCC浸润转移中发挥重要作用，高表达Cyclin A1的HCC患者预后不良；Cyclin A1对判断患者预后具有较高的指导意义。.
Keywords: Cyclin A1; Hepatocellular carcinoma; Invasion; Metastasis; Prognosis.