高表达磷酸丝氨酸转氨酶1抑制胃癌细胞增殖的实验研究

温绍艳; 蔡明志; 梁寒

doi:10.12354/j.issn.1000-8179.2021.20211621

高表达磷酸丝氨酸转氨酶1抑制胃癌细胞增殖的实验研究

doi: 10.12354/j.issn.1000-8179.2021.20211621

温绍艳^1,,
蔡明志²,
梁寒^2, ,

1.
天津医科大学肿瘤医院空港医院综合外一科（天津市300308）
2.
天津医科大学肿瘤医院胃部肿瘤科

基金项目: 本文课题受天津医科大学临床医学研究项目（编号：2017kylc002）资助

详细信息

作者简介:
温绍艳：专业方向为综合肿瘤外科

通讯作者:
梁寒　tjlianghan@126.com

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出版历程
- 收稿日期: 2021-11-15
- 录用日期: 2021-12-10
- 网络出版日期: 2021-12-14

Increased expression of phosphoserine aminotransferase 1 inhibits proliferation of gastric cancer cells

1.
Department of Comprehensive Surgery, Tianjin Medical University Cancer Institute & Hospital, Airport Hospital, Tianjin 300308, China
2.
Department of Gastrointestinal Cancer, Tianjin Medical University Cancer Institute & Hospital, Tianjin 300060, China

Funds: This work was supported by Tianjin Medical University Clinical Research Program (No. 2017kylc002)

摘要

摘要: 目的代谢重塑是肿瘤重要特征之一，多种关键代谢酶在肿瘤发生、发展过程中功能异常。磷酸丝氨酸转氨酶1（phosphoserine aminotransferase 1，PSAT1）催化磷酸丝氨酸生成，促进了肺腺癌、胶质瘤、乳腺癌、结肠癌等肿瘤的恶性发展。本研究分析PSAT1在胃癌中的生物功能。方法利用Kaplan-Meier plotter数据库分析PSAT1与胃癌患者预后的关系；利用RT-qPCR检测PSAT1在胃癌细胞株中的mRNA表达水平；构建PSAT1慢病毒高表达载体，在胃癌BGC823和NCI-N87细胞中高表达PSAT1；通过CCK-8、克隆形成和裸鼠皮下成瘤试验分析PSAT1对胃癌细胞增殖能力的影响。结果 Kaplan-Meier plotter数据库分析显示PSAT1高表达的胃癌患者有更好的总生存期（P=1.7e-6，HR=0.52）。在胃癌KATOⅢ、AGS、SNU16、NCI-N87、MKN45、BGC823、MGC803和SGC7901细胞中，PSAT1的表达水平低于永生化的胃上皮细胞GES1。CCK8试验结果显示PSAT1高表达后可以显著抑制胃癌BGC823（120 h，P<0.000 1）和NCI-N87（96 h，P<0.000 1）细胞的体外增殖能力。克隆形成试验同样证实PSAT1可以抑制胃癌BGC823（P=0.029 6）和NCI-N87（P=0.036 5）细胞的克隆形成能力。进一步动物实验表明PSAT1抑制了BGC823细胞在裸鼠体内的增殖（n=7，P=0.009 1）。结论 PSAT1高表达提示胃癌患者有更好的预后；PSAT1在多数胃癌细胞株中的表达低于GES1细胞；PSAT1高表达可显著抑制胃癌细胞体外、体内的增殖。
- 胃癌 /
- 磷酸丝氨酸转氨酶1 /
- 增殖
Abstract: Objective Reprogrammed metabolism is a hallmark of cancer, and an array of key enzymes are deregulated in various cancer settings. Phosphoserine aminotransferase 1 (PSAT1), which converts 3-phosphohydroxypyruvate to phosphoserine, enhances malignant traits in a few solid cancers including lung adenocarcinoma, glioblastoma, breast cancer, and colorectal cancer. In this study, we investigated the function of PSAT1 in gastric cancer (GC). Methods The Kaplan-Meier plotter database was used to analyze the association between PSAT1 expression and overall survival (OS) rate in patients with GC. PSAT1 mRNA levels were determined using RT-qPCR in a panel of GC cell lines and the immortalized human gastric epithelial cell line GES1. Lentiviral particles expressing PSAT1 were packaged, and then BGC823 and NCI-N87 cells were transduced. The cell counting Kit-8 (CCK-8) and clone formation assay were used to test the role of PSAT1 in the proliferation of GC cells in vitro. Additionally, a xenograft model was established in immunodeficient mice to detect the growth of GC cells in vivo. Results Higher PSAT1 expression indicated longer OS in patients with GC (P=1.7e-6, HR=0.52). PSAT1 mRNA expression in GC KATOIII, AGS, SNU16, NCI-N87, MKN45, BGC823, MGC803, and SGC7901 cell lines was markedly lower than that of GES1 cells. CCK8 assay showed that enforced expression of PSAT1 significantly attenuated proliferation in BGC823 （120 hr, P<0.000 1 and NCI-N87（96 hr, P<0.000 1）cells in vitro . Clone formation assay also demonstrated that PSAT1 overexpression reduced the clone numbers of BGC823 (P=0.029 6) and NCI-N87 (P=0.036 5) cells. Additionally, PSAT1 significantly suppressed growth of the subcutaneously injected NCI-N87 cells in Balb/c nude mice, as evidenced by the reduced tumor weights in PSAT1 group relative to control group (n=7, P=0.009 1) Conclusions The higher expression of PSAT1 indicated favorable prognosis in patients with GC. Moreover, the PSAT1 expression decreased in most GC cell lines compared with that in GES1, and the high expression of PSAT1 markedly inhibited the proliferation of GC cells in vitro and in vivo.
- gastric cancer (GC) /
- phosphoserine aminotransferase 1 (PSAT1) /
- proliferation

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图 1 Kaplan-Meier plotter数据库中PSAT1表达水平与胃癌患者总生存期之间的关系

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图 2 PSAT1在胃癌细胞株及GES1细胞中的表达水平

A：胃癌细胞和永生化胃上皮细胞株中PSAT1的mRNA水平；B：胃癌细胞和永生化胃上皮细胞株中PSAT1的蛋白水平

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图 3 PSAT1在胃癌BGC823和NCI-N87细胞中过表达

A：qPCR检测PSAT1过表达后BGC823和NCI-N87细胞中的mRNA水平；B：免疫印迹实验确认PSAT1过表达后在BGC823和NCI-N87细胞中的蛋白水平

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图 4 PSAT1抑制胃癌细胞增殖能力

A：BGC823细胞；B：NCI-N87细胞

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图 5 PSAT1抑制胃癌细胞克隆形成能力

A：BGC823细胞；B：NCI-N87细胞

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图 6 PSAT1抑制胃癌细胞体内增殖能力

A：对照组和PSAT1高表达组BGC823细胞在裸鼠皮下形成的瘤块；B：对照组和PSAT1高表达组瘤块的重量

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