国际肿瘤学杂志

国际肿瘤学杂志 ›› 2023, Vol. 50 ›› Issue (5): 257-262.doi: 10.3760/cma.j.cn371439-20230302-00052

• 论著 • 上一篇    下一篇

GCSH基因在胃癌SNU-1细胞增殖和凋亡中的作用研究

杨娅1, 王慧礼2, 刘艳1, 郭金凤1, 王春霞1, 吕敏2, 山长平2()   

  1. 1济宁医学院附属医院健康管理中心，济宁 272001
    2济宁医学院附属医院肿瘤科，济宁 272001
  • 收稿日期:2023-03-02 修回日期:2023-04-13 出版日期:2023-05-08 发布日期:2023-06-27
  • 通讯作者: 山长平 E-mail:scp9933@163.com

Effects of GCSH gene on proliferation and apoptosis of gastric cancer SNU-1 cells

Yang Ya1, Wang Huili2, Liu Yan1, Guo Jinfeng1, Wang Chunxia1, Lyu Min2, Shan Changping2()   

  1. 1Health Management Center， Affiliated Hospital of Jining Medical University， Jining 272001， China
    2Department of Oncology， Affiliated Hospital of Jining Medical University， Jining 272001， China
  • Received:2023-03-02 Revised:2023-04-13 Online:2023-05-08 Published:2023-06-27
  • Contact: Shan Changping E-mail:scp9933@163.com

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摘要：

目的 探究敲低甘氨酸裂解系统H蛋白（GCSH）在体外对胃癌SNU-1细胞增殖、凋亡、氧化应激和迁移的影响。方法 体外培养SNU-1细胞并分为对照组（不做转染）、阴性对照组（转染阴性对照siRNA）、GCSH敲低组（转染GCSH siRNA），使用定量PCR方法检测敲低效果，使用免疫荧光观察各组细胞的形态，CCK-8实验检测SNU-1细胞增殖，流式细胞术检测细胞的凋亡水平和氧化应激水平，划痕实验检测细胞迁移。结果 定量PCR实验显示，对照组、阴性对照组、GCSH敲低组的GCSH相对表达量分别为1.29±0.16、1.36±0.17、0.32±0.04（F=90.32，P<0.001），对照组与阴性对照组差异无统计学意义（P=0.497），GCSH敲低组较阴性对照组显著减少（P<0.001）。免疫荧光观察各组细胞的形态无显著差异。CCK-8实验结果显示，对照组、阴性对照组、GCSH敲低组的细胞增殖活力分别为2.63±0.12、2.61±0.14、2.45±0.14（F=6.35，P=0.005），对照组与阴性对照组差异无统计学意义（P=0.751），GCSH敲低组较阴性对照组显著降低（P=0.011）。流式细胞术结果显示，对照组、阴性对照组、GCSH敲低组SNU-1细胞早期凋亡率分别为（13.38±0.45）%、（12.86±0.65）%、（20.04±3.61）%（F=15.37，P<0.001），对照组与阴性对照组差异无统计学意义（P=0.559），GCSH敲低组较阴性对照组显著增加（P=0.002）；3组细胞晚期凋亡率分别为（2.21±0.25）%、（2.68±0.45）%、（5.67±1.67）%（F=18.24，P<0.001），对照组与阴性对照组差异无统计学意义（P=0.356），GCSH敲低组较阴性对照组显著增加（P=0.024）。对照组、阴性对照组、GCSH敲低组活性氧阳性率分别为（26.98±8.79）%、（28.27±5.63）%、（48.41±0.94）%（F=22.56，P<0.001），对照组与阴性对照组差异无统计学意义（P=0.950），GCSH敲低组较阴性对照组显著升高（P<0.001）。对照组、阴性对照组、GCSH敲低组的细胞迁移率分别为（48.29±5.79）%、（51.66±2.29）%、（14.01±1.56）%（F=148.80，P<0.001），对照组与阴性对照组差异无统计学意义（P=0.328），GCSH敲低组较阴性对照组显著降低（P<0.001）。结论 敲低GCSH基因可在体外抑制SNU-1细胞增殖和迁移，增加细胞凋亡率和氧化应激水平，GCSH基因可能是治疗胃癌的潜在靶点。

关键词: 胃肿瘤, GCSH, 细胞凋亡, 细胞迁移

Abstract:

Objective To explore the effects of knocking down glycine cleavage system H protein（GCSH） on proliferation， apoptosis， oxidative stress and migration of gastric cancer SNU-1 cells in vitro. Methods SNU-1 cells were cultured in vitro and divided into control group （no transfection）， negative control group （transfection of negative control siRNA） and GCSH knockdown group （transfection of GCSH siRNA）. Quantitative PCR was used to detect the knockdown effect. Immunofluorescence was used to observe the morphology of cells in each group. CCK-8 was used to test the proliferation of SNU-1 cells. Flow cytometry was used to detect the apoptosis and oxidative stress level， and scratch test was used to detect the cell migration. Results Quantitative PCR experiment showed that the relative expression levels of GCSH in the control group， negative control group and GCSH knockdown group were 1.29±0.16， 1.36±0.17 and 0.32±0.04， respectively （F=90.32， P<0.001）. There was no significant difference between the control group and negative control group （P=0.497）. Compared to the negative control group， the GCSH knockdown group was significantly decreased （P<0.001）. Immunofluorescence experiment showed no significant difference in the morphology of cells among the groups. The CCK-8 experiment results showed that the cell proliferation activities of the control group， negative control group and GCSH knockdown group were 2.63±0.12， 2.61±0.14， 2.45±0.14， respectively （F=6.35， P=0.005）. There was no significant difference between the control group and negative control group （P=0.751）， and the GCSH knockdown group significantly decreased compared to the negative control group （P=0.011）. The results of flow cytometry showed that the early stage apoptosis rates of SNU-1 cells in the control group， negative control group and GCSH knockdown group were （13.38±0.45）%， （12.86±0.65）%， （20.04±3.61）%， respectively （F=15.37， P<0.001）. There was no significant difference between the control group and negative control group （P=0.559）. Compared to the negative control group， the GCSH knockdown group significantly increased （P=0.002）. The late stage apoptosis rates of the three groups were （2.21±0.25）%， （2.68±0.45）%， （5.67±1.67）%， respectively （F=18.24， P<0.001）. There was no significant difference between the control group and negative control group （P=0.356）. Compared to the negative control group， the GCSH knockdown group showed a significant increase （P=0.024）. The reactive oxygen species positive rates in the control group， negative control group and GCSH knockdown group were （26.98±8.79）%， （28.27±5.63）%， （48.41±0.94）%， respectively （F=22.56， P<0.001）. There was no significant difference between the control group and negative control group （P=0.950）. Compared to the negative control group， the GCSH knockdown group significantly increased （P<0.001）. The cell migration rates of the control group， negative control group and GCSH knockdown group were （48.29±5.79）%， （51.66±2.29）%， （14.01±1.56）%， respectively （F=148.80， P<0.001）. There was no significant difference between the control group and negative control group （P=0.328）. Compared with the negative control group， the GCSH knockdown group significantly decreased （P<0.001）. Conclusion Knock down of GCSH gene can inhibit the proliferation and migration， increase cell apoptosis rate and oxidative stress of SNU-1 cells in vitro. GCSH gene may be a potential target for the treatment of gastric cancer.

Key words: Gastric neoplasms, GCSH, Apoptosis, Cell migration