南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (8): 1497-1507.doi: 10.12122/j.issn.1673-4254.2024.08.08

• • 上一篇    

SPAG5在胃癌细胞恶性增殖中的生物学作用

庞一丹1(), 刘雅1, 陈思嫒1, 张荆雷1, 曾今1, 潘元明2(), 安娟1()   

  1. 1.青海大学医学院,青海 西宁 810001
    2.首都医科大学附属北京胸科医院肿瘤研究中心,北京 101149
  • 收稿日期:2024-05-07 出版日期:2024-08-20 发布日期:2024-09-06
  • 通讯作者: 潘元明,安娟 E-mail:15546261023@163.com;peter.f.pan@hsc.pku.edu.cn;anjuan@qhu.edu.cn
  • 作者简介:庞一丹,硕士,E-mail: 15546261023@163.com
  • 基金资助:
    国家自然科学基金(82260846);青海省中藏药管理局科研创新项目(J2022015);青海省科技厅重点研发与转化计划(2021-QY-213)

Biological role of SPAG5 in the malignant proliferation of gastric cancer cells

Yidan PANG1(), Ya LIU1, Siai CHEN1, Jinglei ZHANG1, Jin ZENG1, Yuanming PAN2(), Juan AN1()   

  1. 1.Medical College, Qinghai University, Xining, 810001, China
    2.Cancer Research Center, Beijing Chest Hospital, Capital Medical University, Beijing, 101149, China
  • Received:2024-05-07 Online:2024-08-20 Published:2024-09-06
  • Contact: Yuanming PAN, Juan AN E-mail:15546261023@163.com;peter.f.pan@hsc.pku.edu.cn;anjuan@qhu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(82260846)

摘要:

目的 在胃癌组织及癌旁组织中分析SPAG5的表达情况,通过干扰SPAG5分析其在胃癌细胞生长中的生物学作用。 方法 结合TCGA分析,免疫组织化学、免疫荧光染色分析SPAG5及MKi67在胃癌及癌旁组织中的表达模式,利用胃癌细胞AGS和MGC803进行体外细胞生物学实验,分别设置空白对照组和干扰组,采用慢病毒干扰,其中空白对照转染shCtrl,干扰组转染shSPAG5。通过Celigo、MTT和克隆形成实验、凋亡检测分析敲减SPAG5基因后对胃癌细胞生长的影响。 结果 Proteinatlas数据库、TCGA数据库分析结果发现SPAG5在胃癌中高表达,KM-plot及GEPIA数据库分析SPAG5在肺腺癌、乳腺癌、肝癌、胰腺癌、宫颈癌、膀胱癌中高表达,免疫组化发现SPAG5在胃癌中高表达(P<0.001),免疫组化和免疫荧光共聚焦证明SPAG5与MKi67存在显著相关性(R=0.393,P<0.001)。Real time-PCR及Western blotting结果显示SPAG5在MKN74、BGC823、MGC803、SGC7901和AGS等细胞中表达水平较高(P<0.01)。敲减SPAG5后mRNA和蛋白的表达下降,Celigo、MTT和克隆形成实验结果显示敲减SPAG5抑制胃癌细胞增殖(P<0.01),流式凋亡分析发现敲减SPAG5促进胃癌细胞凋亡(P<0.001)。 结论 SPAG5和MKi67在胃癌组织中的表达具有相关性,干扰SPAG5基因可以抑制胃癌细胞的增殖。SPAG5与患者预后相关,可能成为胃癌的潜在标志物。

关键词: SPAG5, 胃癌, MKi67, 细胞增殖

Abstract:

Objective To analyze the expression of SPAG5 in gastric cancer tissues and its regulatory roles in gastric cancer cell growth. Methods TCGA analysis, immunohistochemistry, and immunofluorescence staining were used to analyze the expression patterns of SPAG5 and MKi67 in gastric cancer and adjacent tissues. In gastric cancer AGS and MGC803 cells, the effects of lentivirus-mediated SPAG5 knockdown on cell growth and apoptosis were evaluated using Celigo, MTT, clone formation assays and flow cytometry. Results Proteinatlas and TCGA database analysis suggested that SPAG5 was highly expressed in gastric cancer, and Kaplan-Meier analysis and GEPIA analysis showed high expressions of SPAG 5 in lung adenocarcinoma, breast cancer, hepatocellular carcinoma, pancreatic carcinoma, cervical cancer and bladder carcinoma. Immunohistochemistry revealed that SPAG5 was highly expressed in gastric cancer tissues (P<0.001), and immunofluorescence colocalization analysis demonstrated a significant correlation between SPAG5 and MKI67 (R=0.393, P<0.001). RT-qPCR and Western blotting showed that SPAG5 was highly expressed in MKN74, BGC823, MGC803, SGC7901 and AGS cells. In AGS and MGC803 cells, SPAG5 knockdown significantly inhibited proliferation and promoted apoptosis. Conclusions The expressions of SPAG5 and MKi67 are correlated in gastric cancer tissues, and SPAG5 knockdown inhibits the proliferation of gastric cancer cells. SPAG5 is associated with the prognosis of gastric cancer patients and may serve as a promising biomarker for gastric cancer.

Key words: SPAG5, gastric cancer, MKi67, cell proliferation