PYCR1对胃癌细胞增殖和侵袭的影响

目的:探讨人吡咯啉-5-羧酸还原酶1（PYCR1）对胃癌细胞AGS增殖和侵袭的影响。方法:人胃癌细胞AGS转染敲减和过表达PYCR1的质粒，设置3种敲减序列，qRT-PCR和Western Blot检测PYCR1 mRNA和蛋白表达以验证敲减效率，并以敲减效率最好的进行后续实验，AGS细胞分为si-NC组、si-PYCR1组、pcDNA组、PYCR1组，采用CCK8检测各组不同时间点细胞活力，采用伤口愈合实验检测各组迁移能力，采用Transwell实验检测各组迁移和侵袭能力。结果:qRT-PCR和Western Blot检测结果发现，三种敲减序列PYCR1 mRNA和PYCR1蛋白均低于si-NC组，过表达PYCR1组PYCR1 mRNA和PYCR1蛋白高于pcDNA组，差异均有统计学意义（P＜0.05）。CCK8检测各组细胞增殖活力，结果发现，si-PYCR1组48 h、72 h和96 h细胞活力显著低于si-NC组相同时间点，而PYCR1组48 h、72 h和96 h细胞活力高于pcDNA组，差异均有统计学意义（P＜0.05）。伤口愈合实验检测各组细胞迁移能力，以48 h/0 h划痕宽度进行定量分析，结果发现，si-PYCR1组48 h/0 h划痕宽度高于si-NC组，PYCR1组48 h/0 h划痕宽度低于pcDNA组，差异均有统计学意义（P＜0.05）。Transwell实验检测各组细胞迁移和侵袭能力，以迁移和侵袭细胞数进行定量分析，结果发现，si-PYCR1组迁移和侵袭细胞数少于si-NC组，PYCR1组迁移和侵袭细胞数多于pcDNA组，差异均有统计学意义（P＜0.05）。结论:PYCR1可以促进胃癌细胞增殖、迁移及侵袭，可能是胃癌发病机制的关键分子和治疗的新靶点。

Effect of PYCR1 on proliferation and invasion of gastric cancer cells

Objective:To investigate the effects of human pyrroline-5-carboxylic acid reductase 1 (PYCR1) on the proliferation and invasion of AGS in gastric cancer cells.Methods:Human gastric cancer cell AGS was transfected with knockdown and over-expressing PYCR1 plasmids.Three kinds of knockdown sequences were set up.qRT-PCR and Western Blot were used to detect the expression of PYCR1 mRNA and protein to verify the knockdown efficiency,and select cells with the best knockdown efficiency for subsequent experiments.AGS cells were divided into si-NC group,si-PYCR1 group,pcDNA group,and PYCR1 group.Cell viability was detected at different time points in each group by CCK8,migration ability of each group was detected by wound healing experiment,and invasion ability of each group was detected by Transwell experiment.Results:The results of qRT-PCR and Western Blot showed that the three knock-down sequences of PYCR1 mRNA and PYCR1 protein were lower than those of si-NC group,and the overexpression of PYCR1 group was higher than that of pcDNA group.The differences were statistically significant (P＜0.05).CCK8 detected the cell proliferation activity of each group,and the results showed that the cell viability of the si-PYCR1 group at 48 h,72 h,and 96 h was significantly lower than that of the si-NC group at the same time.The cell viability of PYCR1 group at 48 h,72 h and 96 h was higher than that of pcDNA group,and the differences were statistically significant (P＜0.05).Wound healing experiments were performed to detect the cell migration ability of each group,and the quantitative analysis was performed with 48 h/0 h scratch width.The results showed that the 48 h/0 h scratch width of the si-PYCR1 group was higher than that of the si-NC group,and the 48 h/0 h scratch width of the PYCR1 group was lower than that of the pcDNA group.The differences were statistically significant (P＜0.05).Transwell test was used to detect the invasion ability of each group of cells,and the number of invaded cells was used for quantitative analysis.The results showed that the number of invasive cells in the si-PYCR1 group was less than that in the si-NC group,the number of invasive cells in the PYCR1 group was more than that in the pcDNA group,and the differences were statistically significant (P＜0.05).Conclusion:PYCR1 can promote the proliferation and migration of gastric cancer cells,and may be a key molecule in gastric cancer pathogenesis and a new target for treatment.